abstract:
Objective To investigate the role of chloride intracellular channels(CLICs) in inflammasome activation duringL.monocytogenesinfection. Methods The primary cultured murine bone marrow macrophages were treated with CLIC inhibitor IAA94 and infected withL.monocytogenes, the levels of interleukin-1 beta(IL-1β) in cell culture supernatants and cell lysates were detected using ELISA and Western blot, respectively. Results The release of IL-1β and lactate dehydrogenase(LDH) decreased in the group of indanyloxyacetic acid 94(IAA94) treatment withL.monocytogenesinfection. Western blot showed that the level of IL-1β in the cell lysates with IAA94 treatment andL.monocytogenesinfection decreased but not in Nod-like receptor family, pyrin domain containing 3(NLRP3) dependent manner. Conclusion Chloride channel proteins is essential in inflammasome activation duringL.monocytogenesinfection, but it may not be in NLRP3-independent way, and the mechanism is still to be further elucidated.

abstract:
Objective To detect the expression changes of pSTAT3 protein in trigeminal ganglions(TGs) of trigeminal neuralgia(TN) model rats and its located cell type. Methods The rat model of TN was established by chronic constriction of infraorbital nerve. Von Frey hair was used to determine the mechanical sensitivity threshold of the two groups, and Western blot was used to detect the changes of pSTAT3 protein expression in TG. Immunofluorescence assay was performed to observe the fluorescence staining of pSTAT3 and its located cell type. Results The CCI-ION group showed a significant reduction of the mechanical pain threshold(P<0.05) from the first day to the fourteenth day after surgery. The expression level of pSTAT3 protein in TG of CCI-ION group was significantly higher than that of Sham group(P<0.05). The immunofluorescence experiment showed that the staining of pSTAT3 in the CCI-ION group was stronger than that in the Sham group, and pSTAT3 was located in microglia cells rather than satellite glia cells or neurons. Conclusion The activation of JAK/STAT3 pathway was found in the TG microglias of TN model rats.

abstract:
Objective To research the expression of β-dystrophin(β-DG) in colon cancer cell lines(HT29,HCT116,SW480,SW620), and the effect of its overexpression on apoptosis and the expression of apoptosis related proteins in SW620. Methods Colon cancer cell lines were cultured, total protein was extracted for Western blot to detect the expression of endogenous β-DG protein in each cell. The eukaryotic expression plasmid of pcDGFP-β-DG was constructed and transfected into SW620 cells, total protein was extracted for Western blot or immunofluorescence directly to detect the overexpression and localization of β-DG. After overexpression of β-DG, the apoptosis of SW620 cells was measured by flow cytometry, and the expression of apoptotic marker protein PARP was further detected. Results Compared with normal epithelial cells HEK 293 T, the expression of endogenous β-DG(43 ku) was decreased in colon cancer cell lines, especially in SW620 cells, and different levels of β-DG hydrolysis fragments(31 ku) were detected. The eukaryotic expression plasmid pcDGFP-β-DG was constructed successfully, which could be effectively expressed in SW620 cells and mainly located in the cytoplasm near the nuclear membrane. After overexpression of β-DG, the apoptotic rate of SW620 cells was(0.167±0.014), significantly higher than that of blank group(0.075±0.027) and control group(0.084±0.023)(P<0.05). After overexpression of β-DG, the protein content of complete PARP(113 ku) decreased, and the fragment of PARP(89 ku) increased. Conclusion The abnormal expression of β-DG in colon cancer cell lines is mainly manifested in its abnormal hydrolytic breakdown. Overexpression of β-DG can significantly promote apoptosis of SW620 cells and expression of apoptotic protein, which provides a cytological basis for further understanding the function of β-DG and its relationship with colon cancer.

abstract:
Objective To investigate the effect of quercetin(Que) on the expression of claudin 6(CLDN6) and the proliferation, migration and invasion of endometriosis cells, and then analyze its possible mechanism. Methods Forty patients with endometriosis were selected. Endometrial tissue was harvested during operation, and ectopic endometrial stromal cells(EcSCs) were cultured. EcSCs were treated with Que at different concentrations(0, 12.5, 25, 50 μmol/L). The effects of different concentrations of Que on the proliferation of EcSCs were detected by MTT assay. The expressions of proliferating cell nuclear antigen and CLDN6 were detected by Western blot. Transwell migration and invasion assays were used to detect cell migration and invasion. Si-control and si-CLDN6 were transfected into EcSCs, and the effects of silencing CLDN6 expression on the proliferation, migration and invasion of EcSCs were analyzed. Subsequent experiments were divided into 4 groups: NC group, Que group, Que+pcDNA group, Que+pcDNA-CLDN6 group. MTT and Transwell migration and invasion assays were used to detect the proliferation, migration and invasion ability of EcSCs. Western blot was used to detect the expressions of proliferating cell nuclear antigen(PCNA), matrix metalloproteinase-2(MMP-2), MMP-9, epithelial-mesenchymal transitions and integrin-linked kinase(ILK) signaling pathway-related proteins. Results The proliferation, migration and invasion of EcSCs were significantly inhibited by Que, which was dose-dependent and significantly inhibited the expressions of CLDN6, PCNA, MMP-2 and MMP-9. Silencing CLDN6 expression could significantly inhibit the proliferation, migration and invasion of EcSCs. Que could significantly inhibit the expression of N-cadherin, Vimentin, ILK and p-AKT proteins and promote the expression of E-cadherin protein. While transfection of CLDN6 overexpression vector significantly reversed the inhibitory effect of Que on proliferation, migration and invasion of EcSCs. Conclusion Que can significantly inhibit the proliferation, migration and invasion of endometriosis cells by inhibiting the expression of CLDN6, which may inhibit the activation of ILK signaling pathway and EMT transition.

abstract:
Objective To investigate the effects of imperatorin on hippocampal injury and atpase activity in rats after cerebral ischemia and reperfusion. Methods The rat model of cerebral ischemia-reperfusion was established,and the rats were randomly divided into 6 groups: healthy control group, model group, imperatorin 25,50,100 mg/kg group and nimodipine group for follow-up experiments. Y maze experiment detected the learning and memory function of rat brain. HE staining was used to detect pathological damage of brain tissue in rats. Apoptosis of rat neurons was detected by TUNEL staining. The contents of total superoxide dismutase(SOD), malondialdehyde(MDA) and glutathione peroxidase(GSH) were detected by the kit. The activity of lactate dehydrogenase(LDH), Na+-K+-ATP and Ca2+-Mg2+-ATP were detected by the kit. Survivin, Caspase-3 and Caspase-9 were detected by Western blot. Results Compared with model group, healthy controls and extraneous imperatorin 50, 100 mg/kg and nim horizon group armed into a significant rise in number of incubation period(P<0.05), the extraneous arms, starting arm into the number, the other arm into number significantly reduced(P<0.05), significantly improved pathological damage degree and the apoptosis positive cells were significantly reduced, Survivin protein level significantly increased(P<0.05), Caspase 3, Caspase-9 proteins significantly reduced(P<0.05), SOD and GSH contents significantly increased(P<0.05), MDA content significantly decreased(P<0.05), LDH activity significantly decreased(P<0.05), Na+-K+-ATP, Ca2+-Mg2+-ATP activity significantly increased(P<0.05). Conclusion Imperatorin has a protective effect on cerebral ischemia and reperfusion in rats.

abstract:
Objective To investigate the effect of hydrogen sulfide(H2S) on proliferation, migration and relaxation of rat basilar artery(BAs) smooth muscle cells(VSMC) and the relationship between H2S and vascular endothelial growth factor receptor 2(VEGFR2). Methods Primary cultured rat BAs VSMC. Specifically, immunofluorescence was used to identify the expression of VEGFR2in BAs VSMC. Cell scratch test was used to observe the change of migration ability of BAs VSMC. CCK-8 method was applied to detect the effect of H2S on the proliferation of BAs VSMC. Cell relaxation test was engaged to detect the relaxation ability of H2S on VSMC. Results The results of cell scratch and value-added experiments showed that, compared with the control group, exogenous H2S donor NaHS(100 μmol/L) could increase the proliferation and migration ability of BAs VSMC and promote VSMC relaxation(P<0.05). In addition, 10 ng/ml VEGF could also enhance the migration, proliferation and relaxation capacity of BAs VSMC, which was similar to the effect of NaHS. On the other hand, pretreatment with 10-5mol/L VEGFR2blocker SU5416 could reduce NaHS cell proliferation, migration and relaxation. Conclusion H2S can promote the proliferation, migration and cell relaxation of VSMC, which may be related to the stimulation of VEGFR2.

abstract:
Objective To explore the expression and purification of PrtC protein fromPorphyromonas gingivalisand its functionin vitro. Methods The whole gene was used to synthesizeprtC gene in ATCC33277 ofPorphyromonas gingivalis, and the recombinant expression vector p29-prtC was constructed,Bioinformatics analysis, low-temperature induced expression,SDS-PAGE and Western blot identification were performed, and the function of collagenase was detected by enzymatic hydrolysis. Results The prokaryotic expression vector of collagenase protein PrtC was successfully constructed, and the soluble protein expression was obtained in the expression system ofE.coli. The protein PrtC with high purity and stability was purified by affinity chromatography and molecular sieve chromatography, and it was proved to have the ability to degrade collagen. Conclusion PrtC protein expressed by prokaryotic recombinant protein had collagenase activity, which laid a foundation for subsequent analysis of PrtC structure.

abstract:
Objective To observe the protective effect of honokiol(HNK) on hydrogen peroxide(H2O2)-induced injury of human neuroblastoma cells(SH-SY5 Y) and explore its possible mechanism. Methods SH-SY5 Y cells were treated with 400 μmol/L H2O2for 24 hours to establish a cell injury model. In the protection group, SH-SY5 Y cells were preincubated with HNK for 6 hours, and then treated with 400 μmol/L H2O2for 24 hours. The activity of SH-SY5 Y cells was determined by CCK8 colorimetry. Apoptosis of SH-SY5 Y cells was determined by flow cytometry. The protein expressions of cytoplasmic Bcl-2, Bax, caspase-3, BIP, CHOP and caspase-12 were detected by Western blot. Results HNK promoted the activity of SH-SY5 Y cells treated with H2O2(P<0.01), decreased cell apoptosis rate, increased Bcl-2/Bax ratio, and decreased protein expressions of Caspase-3(P<0.05), BIP, CHOP and Caspase-12(P<0.01). Conclusion HNK protects SH-SY5 Y cells from H2O2-induced damage by inhibiting ERS-associated cell apoptosis.

abstract:
Objective To investigate the influence on cisplatin resistant of ovarian cancer regulating the expression of miR-214-3 p and analyze the effects of miR-214-3 p on early growth response protein 1(EGR1). Methods Lipofection transfection technology was used for transient inhibition or increasing of miR-214-3 p expression. Cell viability and the half concentration(IC50) of DDP were determined by CCK-8 method. Cell apoptosis was detected by flow cytometry. The mRNA expressions of miR-214-3 p and EGR1, the protein levels of EGR1 and xeroderma pigmentosum(XPD) were measured by using Real-time quantitantive PCR(qRT-PCR) and Western blot, respectively. Results miR-214-3 p in C13 K cells was down-regulated than those in OV2008 cells(P<0.001). After 48 h of transfection, qRT-PCR results revealed that the expression of miR-214-3 p was higher in mimics groups C13 K cells(22.42±4.27) compared to the blank control C13 K cells(0.33±0.16)(P<0.001). The expression of miR-214-3 p in inhibitor groups OV2008 cells(0.11±0.09) was lower than that of blank control OV2008 cells(7.92±2.22)(P<0.001). The decreasing cell proliferation and promoted apoptosis in C13 K cells after transfected with miR-214-3 p mimics was detected(P<0.01), while the increasing of cell proliferation and decreasing of apoptosis in OV2008 cells after transfected with miR-214-3 p inhibitor was detected in the same time(P<0.01). The IC50of DDP in mimics group C13 K cells was decreased, while was promoted in inhibitor group OV2008 cells after transfected(P<0.01). With increased miR-214-3 p expression by mimics transfected, the mRNA and protein expressions of EGR1 greatly increased, and the protein levels of XPD decreased in C13 K cells. By down-regulation of miR-214-3 p, the mRNA and protein expressions of EGR1 greatly decreased, and the protein levels of XPD increased in OV2008 cells. Conclusion miR-214-3 p plays the role of ovarian cancer cisplatin resistant. EGR1 protein is a possible downstream target of miR-214-3 p in ovarian cancer cells.

abstract:
Objective To study the impact of high expression level of HK2 gene in human prostate cancer on T help 1/T help 2,which are co-cultured with human prostate cancer(PC3 cell line). Methods shRNA was used to construct a stable transgenic cells(sh-HK2) of PC3 cell line with shRNA-infected PC3 cell line induced by lentivirus infection, nonsense sequences was used to construct lentivirus, PC3 cell line was infected to obtain negative control cells(NC),then the result of knocking down was inspected through qPCR and Western blot.Isolated cancer cells to obtain supernatant after culturing PC3 cells(NC and sh-HK2) for 48 hours. Human peripheral blood mononuclear cells(PBMCs) were extracted from healthy persons. TH1 blank group: PBMCs were cultured in 1640 medium for 48 h under TH1 induction conditions, and TH1 experimental group(TH1 sh-HK2 group): PBMCs were co-cultured for 48 h in TH1 induction conditions using the supernatant of the cultured stable transgenic cells.(TH1 NC group): PBMCs were co-cultured for 48 h under the condition of TH1 induction using the supernatant of the cultured negative control cells. The induction conditions of the TH2 blank group, the TH2 experimental group(TH2 sh-HK2 group), and the TH2 negative control group(TH2 NC group) were changed to TH2 induction conditions, and the others were unchanged. After 48 hours, the lactate level in each co-culture system was detected by a spectrophotometer. PBMCs were then extracted and cultured with phytohemagglutinin-M(PHA-M) for another 24 h, then the secretion of IFN-γ of PBMCs in TH1 induction conditions and IL-4 of PBMCs in TH2 induction conditions were inspected through ELISA. Results qPCR and Western blot showed that HK2 gene knockdown was significant.In TH1 experimental group of TH1-inducing conditions,the secretion of IFN-γ of PBMCs raised. In TH2 experimental group of TH2-inducing conditions,the secretion of IL-4 of PBMCs decreased. The level of lactic acid in sh-HK2 group declined compared with NC group. Conclusion The high expression level of HK2 gene in PC3 cell line enhances lactic acid in tumor environment. At the same time, it decreases the secretion of IFN-γ and enhances the secretion of IL-4, which leads to weak cellular immune function,therefore humoral immunity that invalid for tumor immunity is enhanced, resulting in the progress of tumor.

abstract:
Objective To investigate the expression and clinical significance of AP-1 complex subunit sigma-1 A(AP1 S1) gene in breast cancer. Methods The relationship between the expression level of AP1 S1 and the clinical prognosis of breast cancer patients was analyzed using the data in surgical tissues and adjacent tissues of breast cancer patients published in the online databases of GEPIA and UALCAN. 110 cases of wax block specimens with complete clinical data and complete clinical data were collected. The expression of AP1 S1 in breast cancer tissues was detected by immunohistochemistry.In vitroexperiments were performed using plate clones, CCK-8 and cell scratch experiments to analyze the effect of AP1 S1 expression on the proliferation and metastasis characteristics of breast cancer cells. Results Through the analysis of GEPIA and UALCAN data, it was found that the expression level of AP1 S1 in breast cancer tissues was significantly higher than that in normal breast tissues(P<0.01). The expression level of AP1 S1 was significantly associated with overall survival(OS) and disease-free survival(DFS)(P<0.05), and it was closely related to the tumor stage of breast cancer patients(P<0.05), and had no obvious correlation with age, tumor classification and lymph node metastasis. Clinical case analysis confirmed that AP1 S1 expression was closely related to breast cancer tumor size and AJCC stage(P<0.05), and had no significant correlation with age, lymph node metastasis, estrogen receptor(ER) status, progesterone receptor(PR) status, and human epidermal growth factor receptor 2(HER2) status(P>0.05). The results of cell function experiments showed that the expression of AP1 S1 gene could promote the proliferation, colony formation and migration ability of breast cancer cells. Conclusion AP1 S1 gene is highly expressed in breast cancer tissues, and its expression level is related to the prognosis of breast cancer patients. Moreover, AP1 S1 gene expression can promote breast cancer cell proliferation, colony formation and migration ability.

abstract:
Objective To prepare Kaposiʼs sarcoma-associated herpesvirus(KSHV) cell antigen tablets and verify their effectiveness, providing a feasible method for the detection of KSHV serum samples. Methods BCBL-1 cells stably expressing KSHV nuclear-associated antigen(LANA) were cultured. BCBL-1 cells were cells infected with KSHV and contained the KSHV gene in their genome. LANA is a protein that is highly expressed on the nucleus of a host cell. When the cells were vigorously viable and in good condition, the cells were collected and fixed to the pre-treated antigen tablets with 4% paraformaldehyde. After blocking,incubation of the serum to be tested and incubation of the fluorescent secondary antibody,the results were observed by immunofluorescence microscopy. Results The fluorescence of the fluorescent secondary antibody was observed in the serum samples of patients infected with KSHV by immunofluorescence technique,while the fluorescence of the secondary antibody was not observed by the specimens not infected with KSHV. Conclusion The cell antigenic tablets of KSHV which can detect serum have been successfully prepared,which provides a method for further studying the epidemiology of KSHV.

abstract:
Objective To compare the related properties of human dental pulp stem cells(DPSCs) and pericytes,and to investigate whether DPSCs can be used to replace pericytes for vascular regeneration. Methods DPSCs and pericytes were induced into osteogenesis and neurogenic differentiationin vitroto identify the multidirectional differentiation potential. Matrigel three-dimensional culture technique was used to evaluate the ability of DPSC and pericytes to stabilize vascular-like structure. Transwell assay was performed to evaluate the induced capacities of DPSCs and pericytes on the migration behavior of human umbilical vein endothelial cell(HUVEC). Results DPSCs and pericytes showed similar morphology and could differentiate into osteogenic and neurogenic cells after induction. The two kinds of cells co-cultured with HUVEC on the Matrigel could establish stable vascular-like structure. In Transwell assay, DPSC and pericytes showed strong induced capacities on the migration behavior of HUVEC. Conclusion DPSCs have similar morphological and functional characteristics to pericytes and can be used for vascular regeneration.

abstract:
Objective To explore effect and mechanism of paeoniflorin-6′-O-benzenesulfonate(CP-25) on human aortic smooth muscle cell(HASMC) with insulin resistance(IR). Methods IR-HASMC was induced by 1 μmol/L insulin. We detected glucose consumption by glucose consumption assay kit. Cell proliferation, migration and apoptosis were detected by CCK-8, transwell and flow cytometry method respectively. The protein expression of GRK2, INSR, p-IRS, PI3 K and GLUT4 were determined by Western blot. The co-expressions of GRK2 and INSR, GRK2 and p-IRS were determined by co-immunoprecipitation. Results CP-25 increased the glucose consumption in IR-HASMC. CP-25 also inhibited abnormal proliferation and migration of cells, and promoted cell apoptosis. CP-25 down-regulated membrane expression of GRK2, up-regulated the expression of INSR, p-IRS, PI3 K and membrane expression of GLUT4. The co-expressions of GRK2 and INSR, GRK2 and p-IRS could be reduced by CP-25. Conclusion CP-25 can improve IR in HASMC, in which it inhibits the membrane translocation and expression of GRK2 and promotes activation of INSR/IRS/PI3 K/GLUT4 signaling pathway.

abstract:
Objective To explore the application of preimplantation genetic haplotyping based on next generation sequencing in process of preimplantation genetic diagnosis of monogenetic diseases. Methods The whole genome of the biopsied trophoblast cells from blastocysts was amplified by multiple displacement amplification method. The pathogenic gene mutation was selected as the target region, and some SNPs were selected as the genetic markers in the upstream and downstream of the target mutation. Preimplantation genetic haplotyping and aneuploidy screening were carried out for embryos by next generation sequencing. The results were confirmed by Sanger sequencing. Results A total of 421 oocytes were obtained from 22 patients, 355 oocytes matured, 305 oocytes fertilized, 301 oocytes cleaved and 143 blastocysts were biopsied. After haplotype analysis, 40 embryos were transplantable, there were 17 transplantation cycles, which lead to 12 clinical pregnancies and 9 live births(1 patient gave birth to 2 infants, which were monozygotic twins). All the infants were followed up in good health. The pregnancy rate was 70.6%. All haplotype analysis results were confirmed by Sanger sequencing and the results were consistent. Conclusion The preimplantation genetic haplotyping method based on the next generation sequencing is feasible, which can help families with high genetic risk to obtain healthy offspring, so as to achieve the goal of eugenics.

abstract:
Objective To investigate the expression of KIAA1199 in pancreatic ductal adenocarcinoma(PDAC) and its clinical significance. Methods 60 PDAC tissues confirmed by pathological diagnosis and 20 pancreatic tissues adjacent to the cancer were collected, and immunohistochemical staining experiments were performed to detect the expression of KIAA1199 gene in paraffin sections. And we analyzed its relationship with related clinicopathological factors and tumor survival time after surgery. At the same time, Western blot and qRT-PCR experiments were used to detect the expression of KIAA1199 in 10 pairs of fresh frozen pancreatic cancer specimens and corresponding adjacent tissues. Results The high expression rates of KIAA1199 in PDAC tissue and adjacent tissues were 70.0%(42/60) and 25.0%(5/20), and the differences were statistically significant(P<0.05). Statistical analysis showed that clinical factors such as lymph node metastasis(P<0.05), differentiation degree(P=0.010), nerve invasion(P=0.016), TNM stage(P=0.047) were closely related to the expression of KIAA1199(P<0.05).However, factors such as age(P=0.206), gender(P=0.515), and tumor location(P=0.726) were not significantly related to the expression of KIAA1199(P>0.05).Using Kaplan-Meier method for survival analysis, the median survival time of KIAA1199 positive overall survival(OS) was 13.5 months and the median survival time of negatively expressed OS was 17 months. The median survival time for disease-free survival(DFS) in KIAA1199-positive patients was 10 months, while the median survival time for DFS in negative-expression patients was 13.5 months. The difference was statistically significant(P<0.05).Western blot and qRT-PCR experiments confirmed that the expression of KIAA1199 in PDAC tissues was higher than that in adjacent tissues, and the difference was statistically significant(P=0.0001,P<0.0001). Conclusion KIAA1199 gene is highly expressed in PDAC tissues, which is closely related to the pathological differentiation of PDAC, TNM stage, nerve infiltration, lymph node metastasis and the prognosis survival time of patients. KIAA1199 may be a new biomarker for predicting PDAC in the future.

abstract:
Objective To explore correlation analysis between movement function and early simple motions in stroke patients' upper limbs at different periods and probability of predictive motion. Methods 102 patients with firstly stroke was observed whether they could perform active finger extension, shoulder abduction and shrug on the 3 rd, 5 th, 7 th, 9 th, and 15 th day after onset. The recovery of upper limb motor function was assessed with action research arm test(ARAT), the modified Barthel index(MBI) and the Fugl-Meyer upper limb subtest(FMA-UE) respectively in the 1 st,3 rd,6 th month after the onset. Results (1) Pearson correlation analysis showed that FE,SA,SS were correlated with ARAT,MBI,FMA-UE(Pearson correlation coefficient: 0.757～0.913,P<0.001).(2) Multiple linear regression showed that SA and SS had no correlation with ARAT, MBI, FMA-UE(P>0.05). There was a correlation between FE of the 3 rd and 5 th day and ARAT, MBI, FMA-UE at the 1 st month after onset(P<0.01); there was a correlation between ARAT and MBI in the 3 th month(P<0.05). FE after 7 th day after the onset was related to ARAT and MBI at 1 st, 3 th and 6 th month(P<0.05), but it was not related to FMA-UE at 3 th and 6 th month(P>0.05).(3) Binary Logistic regression showed that the probability of recovering a certain sensitivity in movement function at 6 th month was 98% in patients with FE of 7 th, and the probability of patients without FE recovering a certain sensitivity in movement function at 6 th month was 38%. The probability of recovering a certain sensitivity in movement function at 6 th month was still 98% in patients who developed FE on 15 th day, but the probability of recovering a certain sensitivity in movement function at 6 th month was 28% in patients without FE. Conclusion (1) SA and SS cannot be used as predictors of motor function after stroke.(2) FE after 7 th days after the onset can be used as a predictor of short, medium and long-term motor function recovery(ARAT, MBI) of affected upper limb in patients with stroke.(3) The later the FE appears on the affected side, the lower the probability of recovery of motor function.

abstract:
Objective To compare clinical effects of mini-stimulation and luteal phase ovulation-inducing programs in patients with failurein vitrofertilization/intracytoplasmic sperm injection(IVF/ICSI). Methods The clinical data of infertile patients with a history of IVF/ICSI failure who received IVF/ICSI treatment at our reproductive center were retrospectively analyzed. They were divided into two groups according to the different stimulation protocols: luteal phase ovulation inducting protocol(98 cases), mini-stimulation protocol(551 cases). The differences in baseline data, ovulation-inducing situation and pregnancy outcome between the two groups were compared. Results There was no significant difference in age, duration of infertility, body mass index(BMI), the basic level of follicle stimulating hormone(FSH), proportion of primary infertility, and proportion of IVF between the two groups(P>0.05); There was also no significant difference in the duration of Gn, the number of retrieved oocytes, MII oocytes, 2 PN zygotes, transferable embryos, and transferred embryos between the two groups(P>0.05). But the total Gn dosage in the luteal phase ovulation-inducing group was significantly higher than that in the mini-stimulation group, and the difference was statistically significant(P<0.05). There was no significant difference in fertilization rate, cycle cancellation rate, implantation rate, and clinical pregnancy rate between the two groups(P>0.05). Conclusion The luteal phase ovulation-inducing and mini-stimulation protocols have similar therapeutic effects in patients with a history of IVF/ICSI failure.

abstract:
Objective To investigate the expression of BTB and CNC homology 1(BACH1) in gastric cancer and its relationship with clinicopathological features and prognosis. Methods Gastric cancer microarray containing 98 samples of gastric cancer with clinicopathological features and follow-up information was used to detect the expression of BACH1 by immunohistochemical method. And the relationship between the expression of BACH1 and the clinicopathological features and prognosis of gastric cancer patients was analyzed. Results The positive expression rate of BACH1 in gastric cancer tissues(44.9%) was higher than that in adjacent normal tissues(25.6%)(P<0.05), which was closely correlated with the pathological grade, T stage, lymph node metastasis and TNM stage of tumor(allP<0.05). K-M survival analysis showed that patients with high expression of BACH1 had a worse prognosis(P<0.05). COX survival analysis showed that the expression of BACH1, lymph node metastasis, distant metastasis and TNM stage were predictors of poor prognosis for gastric cancer(allP<0.05), and the expression of BACH1(P=0.028) and distant metastasis(P=0.001) were independent risk factors for the prognosis of gastric cancer patients. Conclusion The abnormal expression of BACH1 is closely related to the occurrence and development of gastric cancer,which may be a potential biomarker for the prognosis of gastric cancer patients.

abstract:
Objective To investigate the correlation between G3 BP1 and endocrine therapy resistance.Methods Gene Expression Omnibus( GEO) database was retrieved to analyze the expression of G3 BP1 mR NA in breast cancer resistant cell lines. 41 specimens of Luminal-like breast tumor were collected and divided into three groups:primary endocrine resistance group,secondary endocrine resistance group and sensitive group. The expression of G3 BP1 protein in each group was analyzed by immunohistochemical staining. Results GEO database showed the expression of G3 BP1 mR NA in endocrine therapy resistant cell lines was higher than that in sensitive cell lines( P<0. 05). The expression of G3 BP1 protein in primary endocrine resistance group was higher than that in secondary endocrine resistance group( 84. 62% vs 30. 00%,P<0. 01) and sensitive group( 84. 62% vs 28. 60%,P<0. 01). Conclusion G3 BP1 is closely related to endocrine therapy resistance in breast cancer. The high expression of G3 BP1 may lead to primary endocrine resistance.

abstract:
Objective Differential expression of the transcription factor Sp1 was compared in skin tissues and epidermal keratinocytes of psoriasis and normal controls to verify its correlation with the pathogenesis of psoriasis and to explore its clinical significance. Methods Immunohistochemical staining, real-time quantitative PCR and Western blot were used to detect the expression and distribution of Sp1 in mRNA and protein levels in skin tissues of psoriasis vulgaris group and normal control group; After processing Sp1 gene in Hacat cells, the cell proliferation function was detected. Results Immunohistochemical staining showed Sp1 expression was higher in the psoriasis group than that in the normal control group. The psoriasis group was widely expressed in the nucleus of each layer of the epidermis, mainly in the spinous layer and basal layer. Molecular studies showed the expression of Sp1 in mRNA and protein levels of psoriasis group was significantly higher than that of normal control group(P<0.05). After treatment with gene knockdown technology, Hacat cells were subjected to absolute cell counting and CCK-8 counting. The cells in the knockdown group grew slowly and the number of cells decreased, suggesting that Sp1 could promote the proliferation of keratinocytes. Conclusion There may be a correlation between Sp1 and the pathogenesis of psoriasis. The high expression of Sp1 in the skin lesions of patients with psoriasis may participate in the pathogenesis of psoriasis by promoting the proliferation of keratinocytes. At the same time, the expression of Sp1 can provide some reference values for evaluating the biological behavior and treatment effectiveness of psoriasis.

abstract:
Objective To investigate the relationship between vitamin D receptor(VDR) gene polymorphism and susceptibility to pulmonary tuberculosis( PTB) in adolescents,and to provide genetic theoretical basis for prevention and treatment of PTB in adolescents. Methods Peripheral venous blood was collected from 96 patients with pulmonary tuberculosis( as the case group) and 123 healthy controls( as the control group) in Hefei and Shannan.The genotypes of VDR( ApaI,BsmI,FokI,TaqI) were detected by PCR-RFLP,and Chi-square test was used to analyze the relationship between VDR gene polymorphism and TB susceptibility in adolescents from two regions. Results The polymorphism of BsmI locus was significantly different between the case group and the control group in Hefei and Shannan( Hefei: χ2= 19. 948,P<0. 001,Shannan: χ2= 38. 485,P<0. 001). The polymorphism of FokI locus was significantly different between the case group and the control group in Hefei and Shannan( Hefei: χ2=7. 231,P = 0. 027,Shannan: χ2= 21. 781,P<0. 001). But the polymorphisms of ApaI and TaqI locus showed no significant difference between the case group and the control group( P>0. 05). Conclusion VDR gene BsmI and FokI locus polymorphism are related to susceptibility to tuberculosis in adolescents,the Bb,bb,Ff and ff genotypes increase the susceptibility to tuberculosis in adolescents.

abstract:
Objective To investigate the microcirculation alteration in macular area and peripapillary region of myopic eyes and their relationship with axial length(AL) by optical coherence tomography angiography(OCTA). Methods A total of 66 patients(106 eyes) were selected, including 16 patients(28 eyes) in the low myopia group(control group), 23 patients(36 eyes) in the moderate myopia group and 27 patients(42 eyes) in the high myopia group. All patients were performed a complete ophthalmic examination, including spherical equivalent, slit lamp and fundus examination before OCTA inspection, then a series of parameters, including image quality, feveal density 300 μm(FD-300), A-circularity index(AI), parafoveal region superficial capillary plexus(SCP) blood flow density, deep capillary plexus(DCP) blood flow density, peripapillary blood flow density, retinal nerve fiber layer(RNFL) thickness were obtained by using OCTA and the latest version of analysis software. Results There was no significant difference in AI and SCP blood flow density among the three groups(P>0.05). There were statistically significant differences in DCP blood flow density, peripapillary microvascular density and RNFL thickness among the three groups(P<0.05). In the high myopia group and the moderate myopia group, the DCP blood flow density was lower than that of the low myopia group(allP<0.05), while the DCP blood flow density of the moderate myopia group and the high myopia group showed no statistically significant difference(P>0.05). The peripapillary microvascular density of the high myopia group was lower than that of the moderate myopia group and the low myopia group(P<0.05), while there was no statistically significant difference between the peripapillary blood flow density of the low myopia group and the moderate myopia group(P>0.05). Peripapillary blood flow density was positively correlated with RNFL thickness. The blood flow density of DCP, peripapillary vascular density, and FD-300 were all negatively correlated with the axis length. Conclusion Through analysis of OCTA, FD-300,DCP blood flow density and peripapillary microvascular density decrease accordingly with the increase of axis length. Peripapillary microvascular density and RNFL thickness decrease in high myopia.

abstract:
Objective To investigate the expression of lymphocyte subsets, including CD4+T and CD8+T cells immune checkpoint molecules PD-1, LAG-3 and TIM-3 in patients with malignant solid tumors. Methods Flow cytometry(FCM) was used to detect the expression ratios of CD3-cells, CD3+T cells, CD4+T cells, CD8+T cells, CD4/CD8 ratio, B cells, NK cells and their immune checkpoints PD-1, LAG-3 and TIM-3, in peripheral blood of 30 patients with malignant solid tumors and 15 healthy controls. Meanwhile, correlation analysis was performed on lymphocyte subsets and immune checkpoints expression ratios in tumor patients. Results Compared with healthy controls, the proportion of CD3-cells in peripheral blood of tumor patients increased and the proportion of CD3+T cells decreased, but the difference was not statistically significant. Tumor patients compared with healthy controls,there were no statistically significant differences in CD4+T cell ratio(57.64%±15.52%)vs(54.69%±11.33%), CD8+T cell ratio(34.03%±14.27%)vs(31.13%±9.97%), CD4/CD8 ratio 1.94(0.96, 2.68)vs1.85(1.30, 2.67), B cell ratio 5.90%(3.09%,10.69%)vs7.63%(6.67%,10.85%). However, the proportion of NK cells decreased by 8.61%(5.58%, 16.15%)vs21.96%(12.84%, 36.47%). The expression rates of immune checkpoint molecules PD-1, LAG-3 and TIM-3 on CD4+T, CD8+T cells in tumor patients were higher than those in healthy controls, and the proportion of double positive exhausted CD4+T increased, but there was no difference in the proportion of double positive exhausted CD8+T. The proportion of CD4+T cells in tumor patients was negatively correlated with the proportion of PD-1+CD4+T cells and the proportion of double positive exhausted CD4+T cells. Conclusion The proportion of peripheral blood lymphocytes in patients with malignant solid tumors is disordered, negative immune checkpoint molecules are highly expressed and co-expressed, and the proportion of exhausted T cells increased, which negatively regulates cellular immunity. Synergistic effects of immune checkpoint molecules may be involved in the development of tumors. Multi-target combination therapy may yield better results in cancer patients whose immunocheckpoint inhibitors have failed single drug therapy.

abstract:
Objective To compare collateral status on single-phase CT angiography(sCTA) and multiphase CT angiography(mCTA) and their ability to predict a target mismatch on CT perfusion(CTP) and clinical outcomes in patients with acute ischemic stroke(AIS). Methods Sixty-eight AIS patients with occlusions in the M1 segment of the middle cerebral artery and/or intracranial internal carotid artery within 24 h onset were included. The collaterals assessment of sCTA and mCTA were compared. The ability to predict target mismatch on CTP and clinical outcomes was compared between sCTA and mCTA by using receiver operating curve analysis. Results mCTA was better for visualizing collaterals when compared with sCTA. The ability of mCTA to predict target mismatch(AUC=0.82;95%CI:0.727,0.877) and clinical outcome(AUC=0.851;95%CI:0.735,0.944) was better than that of sCTA(P<0.05 overall). When it comes to collateral status>2, mSCTA predicted target mismatch on CTP was the best. The ability of mCTA to predict clinical outcome was comparable to CTP(P=0.29). Conclusion mCTA can be a new tool to combine with CTP for the imaging triage of patients with acute ischemic stroke.

abstract:
Objective To compare the clinical outcomes of double D5, D5+D6 and double D6 blastocysts transfer, meanwhile, explore the effectiveness and safety of D5+D6 blastocysts transfer protocol. Methods A retrospective analysis of 2 180 cycles ofin vitrofertilization/intracytoplasmic sperm injection-embryo transfer(IVF/ICSI-ET) in reproductive medicine center of our hospital. All cycles were divided into three groups in accordance with the different development days of blastocysts: double D5 group(n=1 201), D5+D6 group(n=205) and double D6 group(n=774). Results The biochemical pregnancy rates, clinical pregnancy rates and multiple pregnancy rates in the D5+D6 group were higher than those in the double D6 group(P<0.001), and lower than those in the double D5 group(P<0.001). Double D5 group had higher early pregnancy loss rate compared with D5+D6 group and double D6 group(P>0.05,P<0.05, respectively). The ectopic pregnancy rate in the D5+D6 group was lower than that in the double D5 group and the double D6 group, but there was no significant difference(P>0.05). Conclusion D5+D6 blastocysts transfer as a new protocol can obtain good clinical outcomes.

abstract:
Objective To investigate the correlation between plasma levels of lipoxin A4(LXA4), resolvin D1(RvD1), protectin D1(PD1) and the severity and clinical prognosis of coronary heart disease(CHD).Methods According to the history and clinical examination, 150 patients with CHD were selected and divided into stable angina pectoris(SAP) group, unstable angina pectoris(UAP) group and acute myocardial infarction(AMI) group. Meanwhile, according to the number of coronary artery lesions, 150 patients were divided into single-branch, double-branch and three-branch lesion groups, moreover, the Gensini score was performed. Twenty-six healthy people were selected as the control group. According to whether major adverse cardiovascular events(MACE) occurred during the follow-up, patients with CHD were divided into poor prognosis group and good prognosis group. The levels of plasma LXA4, RvD1 and PD1 were detected, and their correlations with the clinical type of CHD, number of coronary artery lesions, Gensini score and incidence of MACE were analyzed.Results(1) Compared with control group, the levels of plasma LXA4, RvD1 and PD1 were lower in SAP group, UAP group and AMI group(P<0.05). Compared with SAP group, the level of plasma LXA4 in UAP group and the levels of plasma LXA4, RvD1 and PD1 in AMI group were lower(P<0.05), while the levels of plasma RvD1 and PD1 had no statistical significance between SAP group and UAP group(P>0.05). Compared with UAP group, the levels of plasma LXA4 in AMI group were lower(P<0.05), while the levels of plasma RvD1 and PD1 had no statistical significance between UAP group and AMI group(P>0.05).(2) The levels of plama LXA4 and RvD1 in the single-branch lesion group were lower than those in control group(P<0.05), while the level of plasma PD1 had no significant difference between them(P<0.05). The level of plasma LXA4 in the double-branch lesion group was lower than that in control group(P<0.05), the levels of plasma RvD1 and PD1 had no significant difference between them(P<0.05). The levels of plasma RvD1 and PD1 in the three-branch lesion group were lower than those in control group(P<0.05), while the level of plasma LXA4 had no statistical significance between them(P>0.05). The plasma levels of plasma LXA4, RvD1 and PD1 had no statistical significance between single-branch lesion group, double-branch lesion group and three-branch lesion group( P>0. 05).(3) The levels of plasma LXA4,RvD1 and PD1 were negatively related to C-reactive protein and interleukin-6( IL-6)( ρ =-0. 263,-0. 367;-0. 247,-0. 229;-0. 251,-0. 302,P<0. 01),but positively related to IL-10( ρ = 0. 258,0. 245,0. 211,P<0. 01). The levels of plasma RvD1 and PD1 were negatively related to Gensini score( ρ =-0. 227,-0. 229,P<0. 01),however,there was no correlation between LXA4 and Gensini score( ρ =-0. 100,P>0. 05). Stepwise multiple linear regression showed that the levels of plasma LXA4,RvD1 and PD1 had little effect on Gensini score. Stepwise multiple logistic regression showed that plasma PD1 and IL-10 were the protective factors for CHD.(4) Compared with poor prognosis group,the level of plasma PD1 was significantly higher at 3 months of follow-up in the good prognosis group( P<0. 01). Conclusion The level of plasma PD1 can effectively judge the clinical severity of CHD patients,which is a protective factor of CHD and has predictive value for short-term prognosis of CHD patients. The values of plasma LXA4 and RvD1 are weak.

abstract:
Objective To investigate the relationship between the methylation of growth inhibitory factor 4(ING4) gene and cervical carcinogenesis and its clinical significance. Methods Tissue specimens of 80 patients with cervical cancer diagnosed were collected, and 50 cervical tissue specimens from patients with benign uterine lesions undergoing total hysterectomy during the same period were used as the control group. Methylation-specific PCR(MSP) technique was used to detect the methylation status of the ING4 gene promoter in cervical cancer tissues and cervical tissues of the control group, and the mRNA expression level of ING4 gene was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(qRT-PCR). The expression level of ING4 mRNA was analyzed, and the relationship between the methylation status of ING4 gene promoter and the clinical case characteristics of cervical cancer was analyzed. Results The methylation rate of ING4 gene in tissue of cervical cancer(66/80, 82.5%) was significantly higher than that of control group(6/50, 12%)(P<0.01). The mRNA expression of ING4 in cervical cancer tissue(0.41±0.07) was lower than that in control group(0.87±0.16). In addition, the mRNA level in cervical cancer tissues methylated by the ING4 promoter region(0.37±0.06, n = 66) was lower than that in tissues of cervical cancer patients with unmethylated ING4 promoter region(0.46±0.11, n=14). There was no statistically significant difference between the methylation of ING4 gene promoter region and age, lymph node status, pathological classification and clinical stage of cervical cancer in cervical cancer tissues(P>0.05). Conclusion Hypermethylation of the ING4 gene promoter region may be associated with the occurrence and development of cervical cancer.

abstract:
Objective To analyze the influencing factors of postoperative incomplete recovery in patients with type A aortic dissection. Methods A total of 88 patients with type A aortic dissection treated surgically in our depart-ment from January 2018 to November 2019 were selected as the study subjects. According to the postoperative recovery assessment,they were divided into the postoperative incomplete recovery group( n = 52) and the postoperative complete recovery group( n = 36). The body mass index( BMI),extracorporeal circulation( CPB) time,ascending aorta occlusion time,deep hypothermia cycle arrest time,intraoperative cerebral oxygen saturation,history of hypertension,and surgical treatment timing within 24 hours of onset were retrospectively analyzed. Results Univariate analysis showed that there were statistically significant differences in body mass index( P = 0. 002) and CPB time( P<0. 001),time of ascending aorta occlusion( P<0. 001),deep and low temperature shutdown cycle time( P<0. 001),history of hypertension( P = 0. 024),surgery within 24 hours of onset( P = 0. 006),and intraoperative brain oxygen saturation( P<0. 001). For the single factor analysis indicators to be statistically significant in two classifications dependent variable Logistic regression analysis,the results showed that extracorporeal circulation time( OR = 1. 150,P = 0. 001),the cycle time of cryogenic stop( OR = 1. 450,P = 0. 001),brain oxygen saturation( OR = 0. 918,P = 0. 002),hypertension( OR = 0. 682,P = 0. 015),and within 24 hours of surgery( OR = 0. 525,P = 0. 020) were statistically significant independent predictors. Conclusion Extracorporeal circulation time,deep hypothermia shutdown time,intraoperative cerebral oxygen saturation,history of hypertension,and surgery within 24 hours of onset were independent predictors of patients' recovery after type A dissection.

abstract:
Objective By detecting the expression levels of miR-16-5 p and miR-223-3 p in the plasma exosomes of rheumatoid arthritis(RA) patients and normal people, the relationship between them and clinical indicators of RA was analyzed to further understand their clinical value in RA diseases. Methods ExoEasy Maxi kit was used to extract exosomes from RA patients and normal human plasma, and miR-16-5 p and miR-223-3 p were quantitatively detected in exosomes using reverse transcription quantitative polymerase chain reaction technology. Results The expression of exosomes miR-16-5 p in the RA group [1.852(1.084, 2.532)] was higher than that in the healthy control group [0.995(0.572, 1.798)], and the difference was statistically significant(P<0.05). However, its expression level in the RA active group [1.427(0.950, 2.314)] was comparable to the RA stable group [1.970(1.248, 2.857)], and the difference was not statistically significant(P>0.05). The expression level of exosomes miR-223-3 p in the RA group[106.611(46.813, 223.841)] was higher than that in the healthy control group[47.586(8.115, 101.347)], and its expression level in the RA active group [199.615(104.944, 305.090)] was also higher than that in the RA stable group [53.993(21.524, 107.707)](P<0.05). At the same time, the relative expression level of miR-223-3 p was positively correlated with the patients' TJC, SJC, DAS28-CRP and DAS28-ESR scores(P<0.05). Conclusion The abnormal expression of miR-16-5 p and miR-223-3 p in plasma exosomes of RA patients suggests that they may play an important role in the pathogenesis of RA.

abstract:
A retrospective analysis of 258 polycystic ovary syndrome(PCOS) patients and 604 control persons was conducted to analyze the predictive value of uterus thickness on term delivery of PCOS patients and its related factors. Compared with the control group, the uterus thickness of PCOS group was significantly smaller than that of the control group, and the term delivery rate was significantly lower(P<0.05). The area under the ROC curve(AUC) of uterine thickness predicted for the term delivery of PCOS patients was 0.719(95%CI: 0.657-0.781,P<0.001). The cut-off value of uterus thickness was 34.5 mm. There was a significant positive correlation between age and uterus thickness(P<0.001), while there was a significant negative correlation between basal testosterone and uterus thickness(P=0.033) in PCOS patients. The thickness of uterus in PCOS patients is significantly thinner than that in the control group. Uterus thickness is an important factor affecting the term delivery rate of PCOS patients. It has a certain predictive value for the term delivery rate of PCOS patients.

abstract:
In research of dynamic monitoring of the inferior vena cava by bedside ultrasound and using the concept of optimal inferior vena cava width to guide individualized liquid therapy strategies that compared with conventional ICU fluid therapy strategies, it is studied whether it can improve the prognosis of patients with septic shock. Patients with septic shock from the Department of Critical Care Medicine of Anhui Provincial Hospital were admitted to the study. They were randomly divided into two groups by simple randomization method. The control group was treated with conventional treatment strategy while the intervention group was treated with individualized liquid management strategy.The volume and the prognosis of patients in the two groups were contrasted. There was no significant difference in gender, age, underlying disease, infection site, APACHE II score, SOFA score, and mechanical ventilation ratio between the two groups(P>0.05). The cumulative fluid intake, net fluid balance, ICU mortality and the frequency of related complications such as acute heart failure and hypoxemia in the first three and five days of the intervention group were lower than those in the control group(P<0.05).The use of an individualized liquid treatment guided by the concept of the optimal width of inferior vena cava can reduce patients' intake, reduce the risk of complications associated with fluid overload, and improve the patients' prognosis.

abstract:
To study the position of the projection point and the depth of the needle insertion point and depth on the standard or lateral lumbar spine when the lumbar cortical bone screw is implanted, and to provide a theoretical basis for correct nail placement. Ten L1-L5 spine specimens were used to dissect a single vertebral body. The insertion point of the cortical bone channel screw was the intersection of the vertical midline of the corresponding articular process on the corresponding vertebrae and the horizontal line of 1 cm below the ipsilateral transverse process. The open cone was open, the valgus angle was controlled at 10°, and the head tilt angle was 25°-30°. The guide needle was completely implanted into the cortical bone channel to determine the complete nail channel, and the inner sidewall of the pedicle was not damaged. Then the following data were measured and analyzed:(1) The clock recording position of the needle insertion point;(2) In the middle of the pedicle, the position of the needle tip: the projection point on the clock and the distance from the medial edge of the pedicle;(3) At the junction of the pedicle, the position of the tip of the guide pin, the distance from the medial edge of the pedicle;(4) At the position below the upper-end plate, the position of the clock of the guide pin records the position. The results showed:(1) The needle entry point was at 5 o'clock on the left side of the pedicle, and the right side was at 7 o'clock. The needle exit point was on the left side at 11 oʼclock, and the right side was at 1 oʼclock. In the middle part of the pedicle, the tip of the guide needle was close to the inner side of the pedicle in the middle and lower part of the pedicle projection.(3) At the junction of the vertebra and the pedicle, the guide needle was located at the upper pole of the pedicle.