abstract:
Objective To study the effect of mesencephalic astrocyte-derived neurotrophic factor(MANF) on the proliferation and apoptosis of A2 type reactive astrocytes(RAS) in the spinal cord. Methods The spinal cord of SD rats was taken to separate and culture astrocytes, and the cells were identified by immunofluorescence technology. Interleukin-1β(IL-1β) was used to stimulate astrocytes to prepare A2 type RAS as the experimental group; and the untreated group was used as a blank control group. Immunofluorescence was used to identify A2 type RAS. The prepared cells were divided into control group and MANF group. The cells were treated with different concentrations(0, 0.5, 1, 2 μg/ml) of MANF protein and different action times(12, 24, 36,48 h), and CCK-8 method was used to detectthe absorbance at 450 nm. The cells were treated with a certain concentration of MANF protein(1 μg/ml) and divided into the control group and the MANF group. The EdU method was used to verify the effect on the proliferation of A2 RAS. The TUNEL method was used to verify the effect on A2 RAS apoptosis. Results Immunofluorescence results showed that IL-1β could stimulate the transformation of astrocytes to type A2; The results of CCK-8 and EdU proliferation experiments showed that after MANF protein intervention, the cell proliferation ability was significantly higher than that of the control group(P<0.05);TUNEL apoptosis experiment showed that after the intervention of MANF protein, apoptotic cells significantly reduced(P<0.05). Conclusion MANF protein can promote proliferation and inhibit apoptosis of A2 RAS.

abstract:
Objective To study the drug-resistant characteristics of group BStreptococcusin urinary tract infection, the distribution of drug resistance genes and sensitivity of CAMP test and thecfbgene expression. Methods The sensitivity of antimicrobial agents was determined by disk diffusion method and Vitek 2 Compact instrument analysis. The erythromycin induced clindamine resistance was detected by D test. The antibiotic resistance genes were detected by PCR. CAMP test was conducted and strains were identified by detecting 16 S rRNA gene fragments and thecfbgene, biochemical test and MALDI-TOF MS. Results The drug resistance rates of 51 strains of group BStreptococcusto ampicillin, doxycycline, rina thiazole amine, nitrofurantoin, penicillin and quinupristin/dalfopristin, tigecycline, vancomycin were 0, while the drug resistance rates to erythromycin, tetracycline, clindamycin, levofloxacin, moxifloxacin, and ciprofloxacin were 66.7%, 66.7%, 49.0%, 47.1%, 49.0%, and 47.1%, respectively. The main erythromycin and tetracycline resistance genes were ermB and tetM respectively, and quinolones resistance genes were gyrA and parC and their detection rate was all 78.4%. The results of CAMP test showed that 3 strains were negative, 9 strains were weakly positive and 39 strains were positive. Combined with biochemical tests,cfbgene, 16 S rRNA detection and MALDI-TOF MS identification results, all 51 strains were group BStreptococcus. However,cfbgenes of 2 CAMP-negative strains and 2 weakly positive strains were not detected. Conclusion Group BStreptococcusin urinary tract infection is sensitive to a variety of antibiotics, but highly resistant to erythromycin, clindamycin, tetracycline and quinolone antibiotics. There is a certain correlation between drug-resistant gene and drug resistance. CAMP test andcfbgene expression are not sensitive to the identification of group BStreptococcus.

abstract:
Objective To investigate the inhibitory effect of sodium aescinate on bile duct hyperplasia in rats with obstructive jaundice. Methods Eighty one healthy male Sprague Dawley rats were randomly divided into three groups: simple bile duct ligation group(n=36), sodium aescinate group(n=27) and sham operation group(n=18). The first intraperitoneal injection was started 24 hours after bile duct ligation. After continuous administration for 7 days, the liver tissues of rats in each group were obtained. HE staining was used to observe the morphological structure of liver tissues. Masson staining was used to observe the proliferation of collagen fibers in liver tissues. Immunofluorescence staining was used to observe the growth of type Ⅰ collagen fibers(COL-Ⅰ) and type Ⅲ collagen fibers(COL-Ⅲ) in liver tissues of rats in each group. Staining was used to detect the expression of related proteins. Results Sodium aescinate could improve liver fibrosis induced by obstructive jaundice in rats, and inhibit the expression of COL-Ⅰ, COL-Ⅲ and α smooth muscle actin(α-SMA), reduce the proliferation of small bile ducts around liver portal area and the expression of phosphorylated eukaryotic translation initiation factor 4 E binding protein 1(p-4 EBP1) in bile duct cells. Conclusion Sodium aescinate can inhibit the proliferation of small bile duct in rats with obstructive jaundice by reducing the expression of eukaryotic translation initiation factor 4 E binding protein 1(4 EBP1).

abstract:
Objective To investigate the effects of different doses of clozapine on the expression of amylase and pro-inflammatory cytokines [interleukin 1β(IL-1β),interleukin 6(IL-6),and tumor necrosis factor α(TNF-α)] in the serum and pancreatic tissue of male C57 BL/6 mice. Methods Male C57 BL/6 mice were randomly divided into four groups. Mice were treated with clozapine 5,10 and 20 mg/kg or an equal volume of saline by gavage. Mice blood was collected to examine the levels of amylase and pro-inflammatory cytokines( IL-1β,IL-6 and TNF-α) after14 days ofconsecutive gavage. Then the pancreases of the mice were dissected. Western blot was used to detect the expression of amylase,IL-1β,IL-6 and TNF-α protein in pancreatic tissue. Fluorescence quantitative PCR method was used to detect the expression of amylase,IL-1β,IL-6 and TNF-α mRNA in pancreatic tissue. Results Compared with the control group,serum amylase levels significantly increased in the 10 and 20 mg/kg clozapine groups( P<0. 01 and P<0. 001). Compared with the control group,serum IL-1β level of the three clozapine groups increased in a dose-dependent manner( P<0. 01,P<0. 001). Compared with the control group,serum IL-6 level decreased in the clozapine 5 mg/kg group( P<0. 05). Clozapine dose-dependently increased the expression of amylase,IL-1β,IL-6 and TNF-α protein and mRNA in pancreatic tissues of mice,and the differences were statistically significant compared with the normal saline group( P<0. 01). Conclusion Clozapine reliably increased the levels of serum amylase and IL-1β． Clozapine also increased the expression of amylase and pro-inflammatory cytokines IL-1β,IL-6 and TNF-α in pancreatic tissue,which may be one of the pathogenesis of clozapine-induced hyperamylasemia and even pancreatitis.

abstract:
Objective To explore the effect of using Er, Cr: YSGG laser with different power to remove thePorphyromonas gingivalis(P.gingivalis) biofilm on sandblasted, large-grit, acid-etched(SLA) surface. Methods P.gingivalisbiofilm was prepared on SLA surface and randomly divided into three groups. NL: no treatment; L1: 0.75 W Er, Cr: YSGG laser treatment; L2:1.5 W Er, Cr: YSGG laser treatment. Scanning electron microscope(SEM) was used to observe the bacterial biofilm on SLA surface, and live/dead bacterial staining and crystal violet staining methods were used to detect the bacteria in each group. Results Under SEM and live/dead bacterial staining, almost no bacteria was observed in the L1 and L2 groups, but L1 group was still covered with a large amount of acquired pellicle. The results of crystal violet staining showed that L1 and L2 groups were statistically different from NL group, but the difference between the two groups was not statistically significant. Conclusion 1.5 W, 30 Hz Er, Cr: YSGG laser can safely and effectively remove theP.gingivalisbiofilm on the SLA surface.

abstract:
Objective To investigate the effect of autophagy on drug sensitivity of PC9/GR and programmed death ligand-1(PD-L1) in gefitinib-resistant lung adenocarcinoma cellsin vitro. Methods Real-time fluorescence quantitative PCR and Western blot assay were performed to detect the expression levels of autophagy-related proteins LC3 II/LC3 I, Beclin1 and PD-L1 in gefitinib-sensitive cell line PC9 and gefitinib-resistant cell line PC9/GR; CCK-8 assay was performed to detect the sensitivity of PC9 and PC9/GR cells to gefitinib and the inhibition of PC9/GR; The expression of LC3 II/LC3 I, p62 and PD-L1 in PC9/GR cells after autophagy was detected by Western blot. Results The expression of autophagy and PD-L1 was higher in PC9/GR cells than that in PC9 cells, and CCK-8 assay showed that the IC50 for gefitinib was higher in PC9/GR cells than that in PC9 cells, and the sensitivity of PC9/GR cells to gefitinib was partially restored after inhibition of autophagy(P<0.05). PD-L1 expression increased and inhibited autophagy PD-L1 expression decreased with the time and concentration of autophagy inhibitor treatment(P<0.05). Conclusion The results ofin vitroexperiments suggest that inhibition of autophagy may enhance the sensitivity of lung adenocarcinoma resistant cells to gefitinib, suggesting that PD-L1 may be regulated by autophagy and that PD-L1 may be involved in the reversal of gefitinib resistance by inhibition of autophagyin vitro.

abstract:
Objective To investigate the effects and mechanism of knockdown mesencephalic astrocyte-derived neurotrophic factor(MANF) on rifampicin(RFP) induced HepG2 cell injury. Methods A MANF-knockdown stable cell line(MANF Y25) and a control cell line(MANF Y07) were constructed by lentivirus transfection. After HepG2 cells were treated with 100 μmol/L RFP for 24 h, the experiment was divided into MANF Y07+DMSO group, MANF Y07+RFP group, MANF Y25+DMSO group and MANF Y25+RFP group. Western blot and qRT-PCR were used to detect the protein and gene expression levels of MANF and unfolded protein response(UPR)-related genes such as glucose-regulated protein 78(GRP78), protein kinase R-like ER kinase(PERK), eukaryotic translation initiation factor 2α( e IF2α),activating transcription factor 4( ATF4),C/EBP-homologous protein( CHOP),tribbles homolog 3( TRIB3),inositol-requiring enzyme 1( IRE1),spliced X-box binding protein1-S( XBP1-S),non-spliced X-box binding protein 1-U( XBP1-U) and activating transcription factor 6( ATF6); Annexin V-PE/7-AAD double staining was used to detect the apoptosis rate of cells in each group; The changes in the cell proliferation ability were determined by the cell counting kit-8 assay; The relative contents of alanine aminotransferase( ALT),aspartate aminotransferase( AST),alkaline phosphatase( AKP),total bilirubin( TBIL) and indirect bilirubin( IBIL) in the supernatant of cell culture were detected by kits. Results At the protein level,RFP induced the protein expression of MANF,GRP78,p-e IF2α,ATF4 and ATF6; at the gene level,RFP induced the gene expression of MANF and UPR-related genes GRP78,PERK,e IF2α,ATF4,CHOP and TRIB3 in Hep G2 cells. After MANF knockdown,the protein expression levels of GRP78,p-PERK,p-e IF2α,ATF4,ATF6 and the genes expression levels of UPR-related genes mentioned above were further up-regulated( P<0. 05). Moreover,it was also found that after MANF knockdown,the rate of apoptosis increased( P<0. 01),the cell proliferation ability decreased( P<0. 01),and the levels of cell injury markers ALT,AST,AKP,TBIL and IBIL in the supernatant of cell culture increased( P<0. 05). Conclusion RFP activates the UPR,which is further enhanced by MANF knockdown,and cell injury is aggravated,indicating that MANF may play a protective role in RFP-induced Hep G2 cell injury by regulating the UPR.

abstract:
Objective To explore the effect of apigenin on the proliferation and osteogenic differentiation of human dental pulp mesenchymal stromal cells(hDPSCs). Methods CCK-8 method was used to detect the effect of different concentrations of apigenin on the proliferation of hDPSCs; Alkaline phosphatase activity detection, alkaline phosphatase staining(ALP) and alizarin red staining were used to observe the effect of apigenin on the mineralization ability of hDPSCs; Real-time PCR was used to detect the expression of genes related to osteogenic differentiation of hDPSCs under the influence of apigenin.A rabbit skull defect model(4 round defects with a diameter of 8 mm) was constructed and divided into four groups: blank group, bone meal group, normal cultured hDPSCs composite bone meal group, and 5 μmol/L apigenin cultured hDPSCs composite bone meal group.The specimens were taken 4 and 8 weeks after the operation, and the bone volume fraction was measured by Micro-CT. Results CCK-8 showed that 5 μmol/L apigenin had the best effect on the proliferation of hDPSCs; In the 5 μmol/L apigenin group, the results of alizarin red staining showed a significant increase in the number of calcium nodules and a significant increase in alkaline phosphatase activity(P<0.001);PCR results showed that the expression of RUNX2 and OCN was up-regulated in the 5 μmol/L apigenin group.Micro-CT showed that the hDPSCs compound bone meal group cultured with 5 μmol/L apigenin had more new bone formation than the normal cultured hDPSCs compound bone meal group at 4 and 8 weeks, the effect of the blank group and bone meal to form bone was not obvious. Conclusion Apigenin can promote the proliferation and osteogenic differentiation of hDPSCs, providing new ideas for the clinical treatment of periodontal bone defects.

abstract:
Objective To study the effect of Candida albicans with different Sap enzyme activities on the expression of NLRP3 inflammasome in humans and mice vaginal epithelial cells. Methods Candida collected from the vagina of vulvovaginal candidiasis( VVC) patients was purified and identified,and the Sap enzyme activity was detected; Humans and mice vaginal epithelial cells were cultured by enzyme stepwise digestion and co-cultured with Candida albicans with different Sap enzyme activities; The expression of NLRP3 inflammasome was detected in vaginal epithelial cells by immunofluorescence method; The levels of interleukin( IL)-1β and IL-18 in cell supernatant was detected by ELISA method; Cell morphology was observed by HE staining. The culture methods of humans and mice vaginal epithelial cells were in the same way. Results The levels of NLRP3 inflammasome,IL-1β and IL-18 in the group with strong Sap activity were all higher than those in the group with weak Sap activity,and reached their peaks in the early stage of infection. As the infection time increases,cells could be seen to rupture and die,and NLRP3 inflammasomes also declined,this presented the same tendency in humans and mice vaginal epithelial cells. Conclusion Candida albicans could stimulate the expression of NLRP3 inflammasome,IL-1β and IL-18 in vaginal epithelial cells,and Candida albicans with strong Sap activity was more effective. Humans and mice vaginal epithelial cells had the same inflammatory response after infection,so mice could replace humans as the model animal for VVC in vivo research.

abstract:
Objective To research the feasibility of Laponite®(LAP)-sodium alginate(SA) guided bone regeneration composite film used as tissue engineering barrier films by testing its mechanical properties and biocompatibility.Methods Pure SA solution and binary mixture with different LAP and SA ratios(1 ∶10, 3 ∶10, 5 ∶10, 7 ∶10, 9 ∶10) were formed into films through thermal spraying. The tensile strength of the composite films were tested by mechanical universal testing machine(n=6). The microstructure of pure SA film and the composite film with the best tensile strength was observed by scanning electron microscopy(SEM) and transmission electron microscopy(TEM). X-ray diffraction(XRD) and Fourier transform infrared(FTIR) spectrometer were used to characterize the ingredients of the material. Samples with the highest tensile strength was prepared again, followed by immersing in 0.5 mol/L calcium chloride solution for 5 s. The tensile strength of the new group of samples was tested after natural drying. Cell counting kit-8(CCK-8) was applied to detect cell counts in the blank control group, the pure SA film group, the composite film group with the highest tensile strength(n=5) and all groups co-cultured with the NIH/3 T3 for 24 h and 48 h.Results After adding LAP, the cross-section of the composite film showed ordered layer structure. XRD and FTIR showed that LAP could be found in the composite film. The tensile strength first increased and then decreased with the increase of LAP proportion in composite films. The optimal mass ratio of LAP and sodium alginate in composite film was(5 ∶10), and the tensile strength reached(166.9±11.5) MPa before and(178.0±14.8) MPa after Ca2+chelation. The CCK-8 test of NIH/3 T3 showed that there was no significant difference in absorbance between the pure SA film group and the LAP-SA composite film group when they were compared with the blank control group. Conclusion When the mass ratio of the composite film is 5 ∶10, the composite film has the best tensile strengthand good cytocompatibility.

abstract:
Objective To analyze methicillin-resistantStaphylococcus aureus(MRSA) isolated from patients with skin and soft tissue infection(SSTI), and to analyze the homology of staphylococcal cassette chromosome mec(SCC mec), so as to provide laboratory basis for standardizing the rational use of antibiotics and nosocomial infection control. Methods 44 strains of MRSA isolated from patients with SSTI were tested for antimicrobial susceptibility by K-B method and Vitek 2 Compact instrument. MecA gene was detected by ordinary PCR, SCC mec genotyping was performed by multiplex PCR, and homology was analyzed by matrix assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS). Results Mec A gene was detected in all 44 MRSA strains by common PCR detection. Three SCC mec genotypes of Ⅱ, Ⅲ and Ⅳa were identified by multiple PCR, including 5 strains of type Ⅱ(11.36%), 12 strains of type Ⅲ(27.27%), 8 strains of type Ⅳa(18.18%) and 19 strains of untyped type(43.18%). 44 MRSA strains were divided into two clusters by MALDI-TOF MS. Conclusion The main type of MRSA isolated from SSTI patients is SCCmec Ⅲ type. Through the combination of SCCmec genotyping and MALDI-TOF MS technology, it provides a basis for homology analysis among different strains and understands the molecular biology and epidemiological characteristics of MRSA in specific areas.

abstract:
Objective To explore the therapeutic effect of early low-frequency electrical stimulation on knee extension contracture and its possible intervention mechanism in rabbits. Methods Twenty-four New Zealand white rabbits were randomly divided into control group(C), electrical stimulation group(E), natural recovery group(NR) and electrical stimulation treatment group(EST), with 6 rabbits in each group. Group C had free activity for 7 weeks. In group E, free activity was performed for 4 weeks, followed by low frequency electrical stimulation on the quadriceps of the left hind limb for 3 weeks. In group NR, the left knee joint was fixed in the straight position for 4 weeks, and then 3 weeks of free activity was given after the fixed removing. In the EST group, the left knee joint was fixed in the straight position for 4 weeks, followed by low frequency electrical stimulation on the quadriceps of left hind limb for 3 weeks. After each group intervention, the range of motion(ROM) of the left knee joint was measured before and after muscle removal. HE staining and Masson staining were performed on the rectus femurs. The protein expression levels of transforming growth factor-β1(TGF-β1) and α-smooth muscle actin(α-SMA) in rectus and capsule were detected by Western blot. Results Compared with NR group, total contracture and myogenic contracture of the knee joint were reduced(P<0.01) and arthrogenic contracture was not improved in EST group. HE staining of rectus femoris showed that the shape and membrane integrity of muscle fibers in the EST group were improved compared with those of the NR group. The quantitative analysis of masson staining showed that the fibrosis improvement degree of rectus femoris in the EST group was higher than that of the NR group( P<0. 01). In addition,the expression levels of TGF-β1 and α-SMA in the rectus femurs of EST group were lower than those in the NR group( P<0. 05). The expression differences of TGF-β1 and α-SMA in joint capsule of the EST group were not statistically significant compared with those in the NR group( P>0. 05). Conclusion Early lowfrequency electrical stimulation can improve extension contracture of the knee via reducing the level of skeletal muscle fibrosis. The inhibition of skeletal muscle fibrosis is greater than that of joint capsule fibrosis.

abstract:
Objective To investigate the function and mechanism of histone deacetylase 3(HDAC3) in modulating hypoxia-induced autophagy in H9 C2 cardiac cells. Methods The hypoxia model of H9 C2 was establishedin vitro. The expression and fluorescence activity of autophagy-related proteins LC3 B-Ⅱ, Beclin-1, and P62 were detected by Western blot and immunofluorescence staining. After the transfection of HDAC3 plasmid or interfering RNA, the expression and fluorescence activity of autophagy regulatory molecule mTOR and autophagy-related protein LC3 B were detected. Results Compared with the control group, the protein level of LC3 B-Ⅱ and Beclin-1 were augmented after hypoxia for 1 h, 6 h, and 12 h in H9 C2 cardiac cells(P<0.05), and the expression of p62 protein also increased first at 1 h and then decreased at 6 h(P<0.05). After hypoxia for 1 h and 6 h, the protein expression of HDAC3 and mTOR were amplified, while both of them declined at hypoxia for 12 h(P<0.05). The protein level and fluorescence activity of mTOR decreased, but the protein level and fluorescence activity of LC3 B increased after overexpression of HDAC3(P<0.05). Downregulation of HDAC3, the expression and fluorescence activity of mTOR enhanced, while the protein level and fluorescence activity of LC3 B reduced(P<0.05). Conclusion HDAC3 negatively regulates mTOR protein and promotes hypoxia-induced autophagy in H9 C2 cardiac cells.

abstract:
Objective To investigate the effect of hydrogel on oxidative stress and the protective effect of intestinal injury in rats after open abdomen. Methods A poly(ethylene glycol) modified with tyramine was grafted onto a chitosan backbone to enhance the solubility of the chitosan and to crosslink into three-dimensional networks. Followed by intraperitoneal injection of air to induce abdominal compartment syndrome, sixty rats were randomly divided into hydrogel group(CPT group), temporary abdominal closure group(TAC group) and sham operation group(Sham group). Hydrogel group was treated with chitosan-polyethylene-tyramine molecular hydrogel coated on the intestine, and then closed with a polypropylene mesh after open abdomen(CPT group). Temporary abdominal closure group was temporarily closed with a polypropylene mesh after open abdomen(TAC group). Sham group was undergone laparotomies and then closed immediately(Sham group). At immediately four hours after open abdomen,the oxidative stress cells in the small intestinal epithelium of the hydrogel group and the temporary abdominal closure group were detected by immunohistofluorescence. The levels of malondialdehyde( MDA) and 8-OHDG in intestinal homogenate of the three groups were measured at specific time points by TBA and ELISA respectively. Results At 4 h after open abdomen,no oxidative stress injury was observed in intestinal epithelial cells in the CPT group and oxidative stress injury of intestinal epithelial cells was observed in the TAC group. Four hours after open abdomen,MDA and 8-OHDG levels were significantly highest in the TAC group,followed by CPT group,with Sham group was the least. At 8 h and 24 h after open abdomen,MDA and 8-OHDG levels in CPT group returned to Sham values,whereas these levels in TAC group were higher than those of Sham group. Conclusion The biomaterial chitosan-polyethylene glycol-tyramine molecular hydrogels can significantly reduce oxidative stress levels and protect against intestinal damage in small intestine after open abdomen.

abstract:
Objective To investigate whether the ferroptosis inhibitor ferrostatin-1(fer-1) has a protective effect on acute liver injury(ALI) in mice caused by overdose of acetaminophen(APAP). Methods (1) Survival experiment. The 40 male ICR mice were randomly divided into 4 groups, namely APAP model group, fer-1 pre-treatment group, fer-1 post-treatment group, and N-acetyl-L-cysteine(NAC) post-treatment group. They were observed for 7 days after treatment and their deaths were recorded.(2) The 48 male ICR mice were randomly divided into 0 h group, fer-1 group, APAP model group(1, 4, 24 h), and fer-1 pretreatment group(1, 4, 24 h). The pathological changes of mouse liver were observed by HE staining. The aspartate aminotransferase(AST), alanine aminotransferase(ALT) in mouse serum and oxidative stress indicator malondialdehyde(MDA) in liver homogenate were detected by biochemical instrument. RT-qPCR was used to detect the expressions of ferroptosis marker genes long-chain fatty acid-CoA synthase 4(ACSL4), prostaglandin endoperoxidase 2(ptgs2), iron metabolism-related genes transferrin receptor 1(TFR1), hepcidin regulatory protein(HJV), transferrin receptor 2(TFR2) and iron regulatory protein 1(IRP1). Results Compared with NAC post-treatment, fer-1 pretreatment and post-treatment could improve the survival rate of mice; compared with the model group, in the fer-1 pretreatment group, the liver coefficient of mice decreased, liver function indicators ALT, AST improved, lipid peroxidation index MDA decreased, ferroptosis gene expression decreased and iron metabolism disorders were ameliorated. Conclusion Fer-1 pretreatment can alleviate ALI induced by APAP in mice. The mechanism may be through inhibiting ferroptosis and improving the expression of iron metabolism-related genes in the liver of mice.

abstract:
Objective To study the role of presenilin in the reproductive development and oviposition ofSchistosoma japonicum(S.japonicum). Methods The 16, 18, 24 or 28 days post infection(dpi) worms were collected from the mice infected withS.japonicum, then the expression levels of presenilin in the 16, 18 or 24 dpiS.japonicumfemales were analyzed by real-time quantitative reverse transcription PCR(qRT-PCR); The ovaries were collected from 28 dpiS.japonicumfemales, then the expression levels of presenilin in ovary were analyzed by qRT-PCR. The paired schistosomes of 28 dpi were transfected with specific presenilin siRNA and culturedin vitrofor 10 days, then the morphological changes in reproductive organs, male-female pairing and egg production were observed after presenilin gene knockdown. Results The presenilin mRNA expression levels in females 24 dpi were markedly higher than those in females 18 dpi or 16 dpi, in both male and female parasites, higher amounts of the presenilin gene were expressed in ovary tissues. The RNAi of 28 days' pairing worms were treated with siRNA, after 10 days of culturein vitro, the results showed that compared with the control group and the Mock group, the male and female pairing rate of the experimental group decreased significantly; the results of confocal laser scanning microscope showed that in the interference group, the morphology of the reproductive system changed, the number of germ cells decreased and the morphology was abnormal. Conclusion Presenilin plays an important role in the reproductive development and oviposition ofS.japonicum.

abstract:
Objective To study lactoferrin hexapeptide(LfcinB 4-9) to reduce the drug resistance of human ovarian cancer cells and explore its possible mechanism of action. Methods MTT was used to detect the proliferation inhibition of cis-dichlorodiammine platinum(DDP) combined with LfcinB4-9 in human ovarian cancer cells SKOV3, SKOV3/DDP, CI3 K, CI3 K/DDP. The morphologic changes of human ovarian cells were detected by HE staining and section assay. The effect of DDP combined with LfcinB4-9 on the clone formation ability of human ovarian cancer cells was detected by plate cloning assay; Transwell assay was used to detect the effect of DDP combined with LfcinB4-9 on the invasion ability of human ovarian cancer cells. qRT-PCR was used to detect the expression of HSF1, HSP70, OPTN and other genes in human ovarian cancer cells treated with LfcinB4-9 and DDP. Results The results showed that the combined effect of LfcinB4-9 and DDP had a significantly higher inhibitory rate on ovarian cancer cells that of DDP alone(P<0.05 orP<0.01). The cell morphology of human ovarian cells was significantly changed after DDP combined with LfcinB4-9. Compared with DDP alone group, the clone formation ability and invasion ability of human ovarian cancer cells were significantly reduced by DDP combined with LfcinB4-9(P<0.05 orP<0.01). The qRT-PCR assay showed that the expressions of HSF1, HSP70, OPTN and other drug-resistant related genes in DDP combined with LfcinB4-9 were significantly lower than those in the control group and DDP alone group. Conclusion The combined effect of LfcinB4-9 and DDP significantly enhances the sensitivity of ovarian cancer cells to DDP. Its effect is achieved by reducing the expression of HSF1, HSP70, OPTN and other genes through the signalling pathway of HSF1-HSP70-resistant protein.

abstract:
Objective To investigate the growth trajectory of low birth weight infants(LBWI) in infancy, and to establish a growth model of physical development index percentile for LBWI. Methods A total of 261 cases of LBWI were selected in the urban area of Hefei to establish a research cohort. The LMS method was used to establish the percentile curves of age-specific body weight, age-specific length for LBWI infants in Hefei City. Results LMS method was used to fit the body mass and body length curves, and the physical development index percentile curve of LBWI was established. The reference values of body mass and body length percentile of LBWI at different months of infancy were obtained. The fitting effect of growth curve was good. Conclusion This study from the body quality, long established different genders LBWI infantile growth standard reference value and growth curve, to assess LBWI infant growth situation and predict infant disease such as clinical practice to provide the reference and help to understand the Hefei LBWI physique growth characteristics and trajectory, for monitoring LBWI growth and provide reference for evaluation of nutritional status.

abstract:
Objective To study the correlation between occlusal contacts from over-erupted third molar and temporomandibular disorders(TMD). Methods 45 patients with TMD were selected as experimental group: 15 cases(group A) with unilateral mandibular third molar over-eruption, 15 cases(group B) with unilateral maxillary third molar over-eruption, 15 cases(group C) with unilateral mandibular and the contralateral maxillary third molar over-eruption, and 15 cases with normal occlusion as control group. Data of occlusal contacts were recorded with T-scan Ⅲ. Craniomandibular index(CMI), visual analogue scale(VAS) and mandibular motor function before and after the extraction were evaluated. Results In each experimental group, the movement of center of force increased, and the closure time, lateral disclosure time and protrusive disclosure time were longer than those in the control group(P<0.05). The asymmetry index of contact in each experimental group increased(P<0.05). The incidence of premature contact, protrusive interference and lateral interference in each experimental group were significantly higher than those in normal control group(P<0.05), and there was no significant difference in the incidence of premature contact and lateral interference between experimental group and control group after extraction(P<0.05). The CMI and VAS of TMD patients decreased significantly 1 month after the extraction.The maximum vertical opening, protrusive and lateral distance increased(P<0.05). Conclusion Bilateral occlusal symmetry and balance are poor in patients with third molar(s) over-eruption. Occlusal interference caused by the over-erupted third molar(s) is closely related to the clinical symptoms of TMD. Removing the over-erupted third molar(s) can better improve the opening type and vertical opening of the patients with TMD, and relieve the joint noise and pain to a certain extent.

abstract:
Objective To explore the expression of M2 pyruvate kinase(PKM2), cyclooxygenase 2(COX2) and inhibitor of apoptosis protein(IAPs) in cancer tissues of patients with primary liver cancer(PLC) and its relationship with radiotherapy sensitivity. Methods A total of 105 PLC patients who underwent three-dimensional conformal radiation therapy(CRT) were enrolled. Liver cancer tissues were collected by liver needle biopsy. The expression of PKM2, COX2 and IAPs protein families [X-linked inhibitor of apoptosis protein(XIAP), cellular inhibitor of apoptosis protein 1(cIAP-1)] in liver cancer tissues was detected by immunohistochemistry. The differences in expression of PKM2, COX2, XIAP and cIAP-1 in cancer tissues among patients with different radiotherapy effects and sensitivity were compared. The relationship between the expression of above-mentioned proteins and prognosis was analyzed. Results The positive expression rates of PKM2, COX2, XIAP and cIAP-1 in liver cancer tissues of the 105 patients were 81.90%, 74.28%, 85.71% and 82.86%, respectively. There were significant differences in response rate of treatment between patients with positive expression of COX2 and XIAP and those with negative expression(P<0.05). There were significant differences in clinical staging, differentiation degree, tumor diameter, expression of PKM2, COX2, XIAP and cIAP-1 in liver cancer tissues between patients with radiotherapy sensitivity and those without radiotherapy sensitivity(P<0.05). High clinical staging, poor differentiation, positive expression of COX2 and XIAP were independent risk factors of radiotherapy insensitivity(P<0.05). There were significant differences in recurrence rate and metastasis rate between patients with positive expression of XIAP and those with negative expression(P<0.05). Conclusion The positive rates of PKM2, COX2, XIAP and cIAP-1 are high in liver cancer tissues of PLC patients. The expression of COX2 and XIAP is closely related to radiotherapy effect and sensitivity, and the expression of XIAP is also related to recurrence and metastasis.

abstract:
Objective To investigate the changes of serum interleukin-15(IL-15) in patients with knee osteoarthritis(KOA) and healthy controls. Methods The peripheral blood of 30 KOA patients and the peripheral blood of 35 healthy adults were selected. The KOA group was divided into two groups, one was the experimental group with IL-15(10 ng/ml), and the other was without IL-15 as the control group. The HC group was also divided into two groups, one was the experimental group with IL-15(10 ng/ml), and the other was without IL-15 as the control group. ELISA method was used to detect the level of IL-15 in peripheral blood of KOA group and healthy control group. Flow cytometry was used to detect the effect of IL-15 on the expression of natural killer( NK) cell receptor in peripheral blood of patients with KOA and healthy controls,and the ELISA method was used to detect the effect of IL-15 on the levels of tumor necrosis factor α( TNF-α) and interferon γ( IFN-γ) by peripheral blood mononuclear cells( PBMCs). Results The level of serum IL-15 in KOA group was higher than that in control group. IL-15 could enhance the expression of NK cell receptors CD69,CD94 and NKG2 D in KOA group. The levels of TNF-αand IFN-γ in the supernatant of KOA group were higher than those of control group. Conclusion IL-15 antagonist may become a new treatment option to prevent or delay the progression of KOA.

abstract:
Objective To investigate the clinical efficacy of injectable platelet-rich fibrin(i-PRF) combined with vacuum sealing drainage(VSD) technology in the treatment of chronic ulcers. Methods 80 patients with chronic ulcers were enrolled and randomly divided into control group and i-PRF group. In the control group, ulcers was blocked through the negative pressure drainage after wound debridement, and in the i-PRF group, ulcers were conducted multi-point dispersion injection of i-PRF on the wound surface and around the wound after wound debridement, followed by closed negative pressure drainage treatment. All patients repeated the above treatment in a 7-day cycle, and wounds was repaired after reaching the restore standard. The relevant parameters of the two groups were recorded, evaluated and compared on 7 th day and 14 th day after treatment, including bacterial culture, leukocyte, C-reactive protein, erythrocyte sedimentation rate, fresh granulation tissue growth and NRS score, preparation time for the second-stage repair surgery and the time for complete wound healing. Results The inflammation indexes and pain scores decreased in both groups on 7 th and 14 th day after treatment, while i-PRF group showed a more significant downward trend(P<0.05); i-PRF group had higher coverage of fresh granulation tissue and higher rate of bacterial conversion on 7 th and 14 th day after surgery, and the difference was statistically significant(P<0.05); i-PRF group had significantly lower preparation time for secondary repair surgery and complete wound healing time than VSD group(P<0.05). Conclusion Compared with the single application of VSD technology, i-PRF combined with VSD can play a more effective role in treating chronic wounds, controlling infection, reducing pain, accelerating wound healing and reducing hospitalization time.

abstract:
Objective To explore the relationship between serum biomarker's concentrations and early complications after single umbilical cord blood transplantation(UCBT). Methods Concentrations of soluble suppressor of tumorigenicity 2(sST2), regenerating islet-derived protein 3-alpha(REG3α), interleukin(IL)-6, IL-8 and tumor necrosis factor receptor 1(TNFR1) in patients with hematological diseases were dynamically monitored on days 0,7,14,21,28 after UCBT(D0, D7, D14, D21, D28) to analyze the relationship between pre-engraftment syndrome(PES), acute graft versus host disease(aGvHD) and infection and the concentrations of five biomarkers. Results There were no significant differences in biomarker concentrations between PES-/PES+ patients at each time point. D28 sST2 was significantly higher in the aGvHD+ group compared with the aGvHD-group(Z=2.236,P=0.036). There was no statistically significant difference in serum biomarker levels between the two groups at each time point according to whether infection occurred before D28. sST2 showed a moderate or high positive correlation with TNFR1 on D21 and D28(rs=0.633,rs=0.821;P=0.005,P=0.023). Conclusion The elevated serum concentration of sST2 on D28 after single UCBT is closely related to the development of aGvHD.

abstract:
Objective To investigate the risk factors of low anterior resection syndrome after anus-preserving radical resection for rectal cancer and establish a predictive nomogram model. Methods This was a retrospective case-control study. Patients who had undergone anus-preserving radical resection for rectal cancer at department of general surgery of the first affiliated hospital of Anhui medical university completed a LARS score scale. Then the nomogram model was established according to the risk factors of LARS which were assessed by univariate and multivariate analyses. Results Body mass index( BMI) ≥24 kg/m2( OR = 2. 041,95% CI: 1. 038-4. 013),recovery time≤6 months( OR = 2. 456,95% CI: 1. 339-4. 505),the distance from tumor to anus≤7 cm( OR = 2. 735,95%CI: 1. 480-5. 055),neoadjuvant therapy( OR = 3. 772,95% CI: 1. 109-12. 832),anastomotic leak( OR = 5. 537,95% CI: 1. 103-27. 791) were independent risk factors of LARS. Based on the 5 selected risk factors,a nomogram model was established to predict the risk factors of LARS after anus-preserving radical resection for rectal cancer. The area under ROC curve of the nomogram model was 0. 754( 95% CI: 0. 689-0. 819). After internal verification by Bootstrap self-sampling method,the C-index value of the model was 0. 750 and the calibration curve fitted well with the ideal curve. Conclusion The nomogram model based on the above risk factors can better predict the probability of LARS after anus-preserving radical resection for rectal cancer,which is helpful for early identification of hign-risk population and development of clinical intervention measures.

abstract:
Objective To investigate the correlation between tinnitus and insomnia in the elderly people in rural areas of Anhui province. Methods A stratified cluster sampling method was used to survey the 860 elderly people aged 60 and above in rural areas of Huaibei, Hefei, Xuancheng and Anqing. Insomnia Severity Index(ISI) and Tinnitus Handicap Inventory(THI) were used to evaluate the symptoms of insomnia and tinnitus. Results The incidence of insomnia and tinnitus in the elderly people in rural areas of Anhui province were 39.30% and 32.44%. There was a significant difference between the insomnia group and the non-insomnia group(P<0.05). The multivariate regression analysis showed that the elderly people in rural areas with somatic diseases(OR=1.676,95%CI:1.223-2.298) and tinnitus symptoms(OR=1.576,95%CI:1.166-2.132) were more likely to have insomnia. Conclusion The incidence of insomnia and tinnitus is high and tinnitus is a risk factor for insomnia in the elderly people in rural areas of Anhui province, and active measures should be taken to intervene against tinnitus.

abstract:
Objective To compare the efficacy and safety of total body irradiation(TBI) and total bone marrow and lymphoid irradiation(TMLI) combined with chemotherapy for unrelated cord blood transplantation(UCBT) in the treatment of hematological malignancies. Methods Retrospective analysis was performed on 45 patients with hematological malignancies who received UCBT in the affiliated Provincial Hospital of Anhui Medical University. They were divided into TBI group(31 cases) and TMLI group(14 cases) according to different conditioning regimens, in which TBI was performed by electron linear accelerator and TMLI was performed by tomotherapy. The hematopoietic reconstruction, the dose of organs at risk(OAR), the acute and chronic toxic and side effects, and the safety and effectiveness of TMLI dose climbing in the two groups were analyzed and compared. Results The median time of neutrophil engraftment in the TBI group and the TMLI group was 19.50(16-29) days and 16.50(12-38) days respectively(P=0.005), and the cumulative neutrophil engraftment rate by day 42 was 90.32%(95%CI: 88.17%-92.47%) and 100%(95%CI: 97.14%-102.86%)(P=0.025), respectively. The gastrointestinal reaction in the TMLI group was less and the overall treatment time was shorter. Conclusion TMLI provides a new option for patients with refractory leukemia, who cannot tolerate high-dose chemotherapy or cannot adhere to complete TBI. When TMLI dose climbs to 15 Gy, adverse reactions can be tolerated.

abstract:
Objective To investigate the correlation between OLGA and OLGIM staging system and serum pepsinogen(PG) concentrationin patients with chronic atrophic gastritis(CAG), and to analyze the incidence of gastric cancer and its risk factors during follow-up. Methods Endoscopic monitoring was performed on patients who were previously diagnosed as chronic atrophic gastritis(CAG) and were pathologically confirmed to be in line with gland atrophy with or without intestinal metaplasia(IM) and low-grade intraepithelial neoplasia(LGIN). Follow-up patients were endoscopied and biopsied according to the new Sydney system biopsy standards for histological examination,according to the histological results,OLGA and OLGIM staging were performed. Stage 0 to II were low risk of gastric cancer,and stage Ⅲ to Ⅳ were high risk of gastric cancer. Serum pepsinogen and gastrin-17 were measured with ELISA methods at the same time. Results 164 patients with atrophic gastritis were enrolled,averaging age of( 57. 48 ± 10. 71) and male/female 102/62. All patients had been diagnosed with gastric mucosal atrophy with or without intestinal metaplasia and low-grade intraepithelial neoplasia,and the mean follow-up time was( 4. 2± 3. 3) years at least two endoscopic follow-ups were performed. Compared with the low-risk groups with OLGA stage and OLGIM stage,G-17 was significantly increased in the high-risk group,PGI and PGR were significantly decreased,and the difference was statistically significant( P<0. 05). During the follow-up,6 cases of gastric cancer were detected( 3. 65%),and 4/2 were male/female. Among them,there were 5 cases of early gastric cancer and 1 case of advanced gastric cancer,and the tumor progression rate of chronic atrophic gastritis was0. 58% annually. COX regression analysis showed that patients older than 60 years old with atrophic gastritis and low-grade intraepithelial neoplasia were risk factors for the prediction of gastric cancer of atrophic gastritis. Conclusion Chronic atrophic gastritis is a precancerous condition of gastric cancer,and endoscopic follow-up is helpful for early detection of gastric cancer. For patients over 60 years old with severe atrophic gastritis and low-grade intraepithelial neoplasia,it is recommended to reexamine high-definition gastroscopy every 1-2 years.

abstract:
Objective To investigate the differences in gray matter volume(GMV) changes of male and female patients with Alzheimer′s disease(AD) and the potential correlation with cognitive impairment. Methods Eighty-eight patients with AD(37 males and 51 females), 84 patients with amnestic mild cognitive impairment(aMCI, 40 males and 44 females) and 79 healthy controls(HC, 34 males and 45 females) were included. High-resolution 3-dimensional T1 structure images were obtained from each participant. The optimized voxel-based morphological measurement(VBM) method and two-way ANOVA analysis were conducted to investigate differential brain regions with interaction between groups and sexes, which were then chosen as regions of interest(ROI) to extract GMV by the REST software for correlation analysis with the Mini-mental State Examination(MMSE) scores. Results The right temporal pole, right orbital inferior frontal gyrus, right middle temporal gyrus, bilateral anterior cingulate cortex and right middle cingulate cortex were brain regions with interaction between sexes and groups of AD, aMCI and HC(P<0.05,FWEcorrected at cluster level). In HC and aMCI groups, the GMV of females in the aforementioned areas was similar to or slightly larger than that of the males, while in the AD group, it bacame significantly smaller than that of males. For correlation analysis, the aMCI and AD patients were combined and then dimidiated into two groups by sex, MMSE scores of the female patient group positively correlated with GMV of the aforementioned five brain regions(P<0.05), MMSE scores of male patient group were positively correlated with GMV of the right middle temporal gyrus(r=0.265,P=0.020). The aMCI and AD patients were separated and then dimidiated into two groups by sex, MMSE scores positively correlated with the GMV from the right middle temporal gyrus in both the female AD group and male aMCI group(r=0.327,P=0.019 andr=0.419,P=0.007). Conclusion There is a difference between the sexes in terms of gray matter atrophy while AD progresses. In particular, some brain regions atrophy more severely in female patients. The different gray matter atrophy patterns in males and females may lead to differences in cognitive decline during the AD disease process.

abstract:
Objective To investigate the risk factors and etiological characteristics of urinary tract infection in continuous ambulatory peritoneal dialysis(CAPD) patients. Methods A total of 90 CAPD patients with urinary tract infection in the past 5 years were selected as the infection group. In addition, 36 patients with CAPD without urinary tract infection were selected as the control group. The clinical characteristics of the two groups were compared. The risk factors of urinary tract infection and the etiological characteristics were analyzed. Results A total of 90 strains of bacteria were isolated from the urine culture of 90 CAPD patients with urinary tract infection.Escherichia coli,Enterococcus faecalisandKlebsiella pneumoniaewere the most common bacteria. The proportion of women in the urinary tract infection group was 84.4%, which was higher than 38.9% in the control group(χ2=26.149,P<0.001). The proportion of people with 24-hour urine volume less than 200 ml was about 50% in the infection group, which was higher than 16.7% in the control group(χ2=11.859,P<0.05). The peritoneal dialysis age, total cholesterol, triglycerides, fasting blood glucose, blood calcium, calcium and phosphorus product of the urinary tract infection group were higher than those of the control group(P<0.05). Multivariate binary logistic regression analysis indicated that female was an independent risk factor for urinary tract infections(OR=6.218,P=0.001). Conclusion Urinary tract infections in peritoneal dialysis patients are more common with Gram-negative bacteria. The urinary tract infection group has the characteristics of long dialysis age, higher total cholesterol, triglycerides, fasting blood glucose, blood calcium, calcium and phosphorus product, and low urine volume over 24 hours compared to the control group. Female is an independent risk factor for urinary tract infections.

abstract:
Objective To investigate the effect of atosiban on frozen-thawed embryo transfer outcomes in infertile patients with scar uterus. Methods This was a retrospective study.278 secondary infertile patients with cesarean scar uterus were divided into two groups, the study group(n=128) treated with low-dose atosiban(6.75 mg/0.9 ml, iv) within 30 min before fozen-thawed embryo transfer, while the control group(n=150) without treatment with atosiban.Then the two groups were respectively stratified according to the times of embryo transfer, which were the first time, second time and third time or more.The differences of embryo implantation rate, biochemical pregnancy rate, clinical pregnancy rate and live birth rate between study group and control group were compared. Results The embryo implantation rate, biochemical pregnancy rate, clinical pregnancy rate and live birth rate of the overall study group were improved, but the difference was not statistically significant(P>0.05); atosiban could greatly improve the transfer outcomes of patients in the second embryo transfer, but the difference was not statistically significant(P>0.05). Conclusion Treatment with atosiban before frozen-thawed embryo transfer can′t significantly improve transfer outcomes of infertile patients with scar uterus.

abstract:
Extraction of CP-PGN was processed according to the TCA method, the purity of the final product was verified by treating with the lysozyme. Then the molecular weight, structures and compositions of amino acids of CP-PGN were studied. The animal models of bloodstream infection were established by using carbapenenes producingKlebsiella pneumonia(KPN) andCandida albicans(CAL) to the evaluation of the effect of CP-PGN against bloodstream infection. The lysozyme dissolution experiment verified that the extracted product was peptidoglycan and CP-PGN possessed the characteristics of peptidoglycan. The molecular weight of CP-PGN is 18 774 u, which showed the characteristic peaks of PGN by infrared spectroscopy. The GB/T14965-1994 was used to detect its compositions of amino acids. CP-PGN significantly prolonged the survival time, improved the survival rate of mice and showed different degree of prevention in the animal experiments. All these showed that the extracted product of CP-PGN had the significant preventive and resistant effects on KPN and CAL-induced bloodstream infection.

abstract:
Cytomics FC500 flow cytometer and ImageStreamX Mark Ⅱ imaging flow cytometer were used to detect the percentage of mouse-derived RAW264.7 cells transfected with control group NC mimics and microRNA-22(miR-22) mimics to phagocytose mouse-derived glioma cells GL261, and the correlation and consistency of the two detection technologies were observed. The results showed that the ratio of RAW264.7 cells(NC mimics and miR-22 mimics) phagocytosing GL261 cells measured by two kinds of flow cytometry had a good correlation(R2>0.8). The Bland-Altman method analyzed the two detection techniques and found that the proportion of RAW264.7 cells(NC mimics and miR-22 mimics) phagocytosing GL261 cells was less biased, which were 4.289% and 8.073%, respectively. Compared with the control group NC mimics, the ability of RAW264.7 cells in miR-22 mimics group to swallow tumor cells increased(P<0.05), and imaging flow cytometry could also obtain individual cell pictures of the phagocytic process. Classical flow cytometry is easy to operate and fast while imaging flow cytometry is based on traditional flow cytometry, which realizes multi-parameter quantitative analysis of cell images and can obtain new statistical data of cell morphology.

abstract:
To investigate the CT features of lung adenocarcinoma with ground glass nodules(GGN) on thin slice CT(TSCT) and to compare with the pathological classification. A total of 83 pulmonary ground glass nodules were collected retrospectively. According to the pathological results, the lesions were divided into pre-invasive lesions group with 20 lesions, microinvasive adenocarcinoma(MIA) group with 32 lesions, and invasive adenocarcinoma(IAC) group with 31 lesions. The appearance of nodules with different pathological types on thin-slice CT was analyzed. Among 83 lesions, 34 1 esions were pure ground glass opacity(pGGN),while 49 lesions were mixed ground glass opacity(mGGN).pGGN was observed in pre-invasive lesions group(18/20,90.0%), MIA group(16/32,50.0%) and IAC group(0/31,0%), respectively. Among the 3 groups, the proportion of foliation sign, air bronchi sign, vascular cluster sign and pleural depression sign showed an increasing trend, while the proportion of round/quasi-round sign and clear sign of tumor lung interface showed a decreasing trend. There were statistically significant differences in round or quasi-round, lobulated sign, clear lung interface of tumor, air bronchi sign, vascular cluster sign and pleural depression sign(P<0.05). There were statistically significant differences between pre-invasive lesions group and IAC group in round or round shape, lobulation sign, clear lung interface of tumor, air bronchi sign, vascular cluster sign and pleural depression sign(P<0.05). There were statistically significant differences between MIA group and IAC group in terms of roundness, lobulation sign, clear lung interface of tumor and vascular cluster sign(P<0.05). The manifestations of GGN of different pathological types on TSCT have certain specificity, and familiarity with the manifestations of TSCT is conducive to the diagnosis and differential diagnosis of pre-invasive lesions, micro-invasive adenocarcinoma and invasive adenocarcinoma.