Objective To explore the repair effect and mechanism of human amniotic mesenchymal stem cells(hAMSCs)on endometrial injury.Methods hAMSCs were isolated using a two-enzyme digestion and then cultured.The third-passage(P3)cells were harvested to detect the surface markers by flow cytometry and to identify their trilineage differentiation potentials.Eighteen nulliparous female SD rats(8～9 weeks old,weighing 250～280 g)were randomly divided into 3 groups(n=6):normal control group,model group,and hAMSCs group.A rat model of intrauterine adhesions(IUA)was established in SD rats by using curettage combined with lipopolysaccharide(LPS)infection.In 2 weeks after modeling,the hAMSCs group received a bilateral uterine horn transplantation of 0.2 mL hAMSCs(1.0×10? cells/mL),while the model group received a same volume of PBS into both uterine horns.All rats were sacrificed in 2 weeks after transplantation.HE and Masson staining was used to observe endometrial thickness and gland number as well as endometrial fibrosis area.RT-qPCR and Western blotting were performed to detect the mRNA and protein levels of TGF-β1,Smad3,Smad7,epithelial-mesenchymal transition(EMT)markers(E-cadherin,Vimentin),fibrosis factor α-SMA,and endometrial estrogen receptor(ER)and progesterone receptor(PR)in endometrial tissues.Results The obtained cells were identified as hAMSCs due to the characteristics of surface markers and differentiation potentials.Compared with the normal control group,the model group showed decreased endometrial thickness,reduced gland number,increased fibrosis area,and enhanced mRNA and protein levels of fibrosis-related factors TGF-β1,Smad3,Vimentin,and α-SMA(P<0.01),while down-regulation of fibrosis-inhibiting molecule Smad7,the EMT marker E-cadherin,and endometrial receptors ER and PR at both mRNA and protein levels(P<0.01).hAMSCs transplantation increased endometrial thickness and gland number,decreased fibrosis area,and down-regulated mRNA expression of the aforementioned fibrosis-related factors(P<0.01),and up-regulated the mRNA expression levels of Smad7,E-cadherin,ER,and PR(P<0.01).The hAMSCs group also exhibited obviously down-regulated protein levels of TGF-β1,Smad3,and α-SMA(P<0.05),while enhanced protein levels of Smad7 and PR(P<0.05).Conclusion Intrauterine transplantation of hAMSCs can promote the repair of endometrial injury,and inhibits endometrial EMT and fibrosis through the TGF-β1/Smad7 signaling pathway.

OBJECTIVE To study the effects of Zigui yichong formula on premature ovarian insufficiency （POI） in mice through glycolysis metabolic pathway. METHODS Eighty SPF C57BL/6N female mice were divided into normal group， model group， Zigui yichong formula group （14.175 g/kg）， Zigui yichong formula+2-deoxy-D-arabino-hexose （2-DG） group （Zigui yichong formula 14.175 g/kg + glycolysis inhibitor 2-DG 100 mg/kg）， with 20 mice in each group. Except for the normal group， POI model mice were induced by intraperitoneal administration of cyclophosphamide in the other groups. After the model was successfully established， each group was given corresponding drugs. HE staining was employed to observe the pathomorphological changes in ovarian tissue and to count follicles at all developmental stages； radioimmunoassay was conducted to measure the serum levels of estradiol （E2）， anti-Müllerian hormone （AMH）， and follicle-stimulating hormone （FSH）； TUNEL assay was employed to detect apoptosis in ovarian granulosa cells of mice； the activities of hexokinase （HK）， pyruvate kinase （PK） and lactate dehydrogenase （LDH） were detected by colorimetry； Western blot and real-time fluorescence quantitative PCR were employed to analyze the protein and mRNA expressions of B-cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， caspase-3， HK2， pyruvate kinase M2 （PKM2）， and lactate dehydrogenase A （LDHA）. RESULTS Compared with model group， the number of primordial follicles， growing follicles， antral follicles and granulosa cells were increased significantly（P＜0.05）， and granulosa cells arranged neatly， but the number of atretic follicles and granulosa cells apoptosis were decreased significantly in Zigui yichong formula group （P＜0.05）； the serum levels of E2 and AMH， the activities of HK， PK and LDH， protein and mRNA expressions of Bcl-2， HK2， PKM2 and LDHA were increased significantly （P＜0.05）； the serum levels of FSH， the protein and mRNA expressions of Bax and caspase-3， Bax/Bcl-2 ratio were decreased significantly （P＜0.05）. 2-DG could reverse the improvement effects of Zigui yichong formula on the above indexes of POI model mice. CONCLUSIONS Zigui yichong formula may inhibit the apoptosis of ovarian granulosa cells， reduce follicle atresia and improve ovarian reserve function by promoting glycolysis levels in POI model mice.

Objective:To investigate the effect of human amniotic mesenchymal stem cells (hAMSCs) on ovarian function and endometrial receptivity in mice with autoimmune premature ovarian insufficiency (POI).Methods:POI mouse was induced by treatment with zona pellucida 3 polypeptide fragment-Freund immune adjuvant. The animals were divided into normal group ( n=10), model group ( n=15) and hAMSCs group ( n=15). hAMSCs (1×10 6 cells/mouse) were transplanted by tail vein single injection. The oestrus cycles were evaluated by vaginal smears. The levels of follicle-stimulating hormone (FSH), estrogen and anti-Müllerian hormone (AMH) in serum were detected by enzyme-linked immunosorbent assay methods. Morphological changes of ovarian and uterus tissues were observed after HE staining. The expressions of homeobox A10 (HOXA10), tumor necrosis factor-α (TNF-α) and FSH receptor (FSHR) proteins in uterine were measured by immunohistochemistry. The endometrial receptivity was comprehensively assessed. Results:After hAMSCs transplanting 6 weeks, the rate of abnormal oestrus cycles in hAMSCs group [40.0% (6/15)] was lower than that in model group [86.7% (13/15), P=0.021]. Compared with the levels of serum FSH [(10.239±1.091) μg/L], estradiol [(103.325±4.952) ng/L] and AMH [(1.133±0.494) μg/L] in model group, the level of FSH in hAMSCs group [(7.664±0.735) μg/L] was significantly decreased ( P<0.001), the levels of estradiol [(126.883±23.370) ng/L] and AMH [(2.204±0.453) μg/L] were significantly increased in hAMSCs group ( P=0.015, P<0.001). Different from model group, the ovarian and uterine index were increased. A large number of healthy follicles at all stages were highly increased, but it was rare to find interstitial fibrosis and atresia follicles. The uterine wall and endometrium were thickened, and the number and volume of the glands were increased. The absorbance ( A) of HOXA10 in hAMSCs group (5.90±1.94) was higher than that in model group (2.79±1.27, P=0.029). The TNF-α A value of hAMSCs (3.83±1.23) group was significantly lower than that of model group (6.26±0.96, P=0.002). Although there was no significant difference on FSHR A value between hAMSCs group (3.61±1.66) and model group (2.74±0.22, P>0.05), the FSHR A value of model group was lower than that of normal group (4.13±0.54, P=0.006). Conclusion:hAMSCs transplantation could restore ovarian function of autoimmune POI mice meanwhile significantly improve uterine receptivity and fertility.

Objective:To investigate the effect of human amniotic mesenchymal stem cells (hAMSCs) on ovarian function and endometrial receptivity in mice with autoimmune premature ovarian insufficiency (POI).Methods:POI mouse was induced by treatment with zona pellucida 3 polypeptide fragment-Freund immune adjuvant. The animals were divided into normal group ( n=10), model group ( n=15) and hAMSCs group ( n=15). hAMSCs (1×10 6 cells/mouse) were transplanted by tail vein single injection. The oestrus cycles were evaluated by vaginal smears. The levels of follicle-stimulating hormone (FSH), estrogen and anti-Müllerian hormone (AMH) in serum were detected by enzyme-linked immunosorbent assay methods. Morphological changes of ovarian and uterus tissues were observed after HE staining. The expressions of homeobox A10 (HOXA10), tumor necrosis factor-α (TNF-α) and FSH receptor (FSHR) proteins in uterine were measured by immunohistochemistry. The endometrial receptivity was comprehensively assessed. Results:After hAMSCs transplanting 6 weeks, the rate of abnormal oestrus cycles in hAMSCs group [40.0% (6/15)] was lower than that in model group [86.7% (13/15), P=0.021]. Compared with the levels of serum FSH [(10.239±1.091) μg/L], estradiol [(103.325±4.952) ng/L] and AMH [(1.133±0.494) μg/L] in model group, the level of FSH in hAMSCs group [(7.664±0.735) μg/L] was significantly decreased ( P<0.001), the levels of estradiol [(126.883±23.370) ng/L] and AMH [(2.204±0.453) μg/L] were significantly increased in hAMSCs group ( P=0.015, P<0.001). Different from model group, the ovarian and uterine index were increased. A large number of healthy follicles at all stages were highly increased, but it was rare to find interstitial fibrosis and atresia follicles. The uterine wall and endometrium were thickened, and the number and volume of the glands were increased. The absorbance ( A) of HOXA10 in hAMSCs group (5.90±1.94) was higher than that in model group (2.79±1.27, P=0.029). The TNF-α A value of hAMSCs (3.83±1.23) group was significantly lower than that of model group (6.26±0.96, P=0.002). Although there was no significant difference on FSHR A value between hAMSCs group (3.61±1.66) and model group (2.74±0.22, P>0.05), the FSHR A value of model group was lower than that of normal group (4.13±0.54, P=0.006). Conclusion:hAMSCs transplantation could restore ovarian function of autoimmune POI mice meanwhile significantly improve uterine receptivity and fertility.

Objective To study the association between serum TB level and target organ damage in elderly metabolic syndrome (MS) patients.Methods Two hundred and forty-five elderly MS patients admitted to our hospital were included in this study.Their general condition was recorded,their serum TB,blood glucose,blood lipids,blood urea nitrogen and creatinine (Cr) levels were measured,and their left ventricular mass (LVM) and left ventricular mass index (LVMI) were detected by parallel echocardiography.Results Correlation analysis showed that the serum TB level was positively related with that of Cr (r=0.168,P=0.009) but not related with LVM,LVMI,serum blood urea nitrogen level,prevalence of chronic renal insufficiency and chronic kidney disease (P＞0.05).Regression analysis showed that serum TB level was an independent risk factor for urea in MS patients (OR=-0.27,95％CI:-0.48-0.06,P=0.01;OR=1.27,95％CI:0.33-2.22,P=0.01).Quantile analysis of serum TB level showed that the serum Cr level was significantly higher in Q76-100 group,Q51-75 group and Q26-50 group than in Q1-25 group (P=0.031).Conclusion Serum TB level is positively related with endogenous Cr clearance in elderly MS patients,suggesting that mildly elevated serum TB level may be a protective factor for impaired renal function in elderly MS patients.

Objective To label granulocytes in a state of differentiation in mouse bone marrow (BM) with EdU (5-ethynyl-2′-deoxyuridine) for further understanding the changes in granulocyte produc-tion at different stages of differentiation during inflammation. Methods C57BL/6 mice were intraperitoneal-ly (i.p.) injected with EdU and heat-inactivated Escherichia coli(HI E.coli). BM cells were harvested at different time points after HI E.coli injection and then stained with fluorescent-conjugated antibodies(Abs). Myeloblasts,promyelocytes,myelocytes, metamyelocytes and band and segmented neutrophils were identi-fied by fluorescence-activated cell sorting(FACS). The percentage of EdU-positive cells in each population was recorded. Results The percentage of EdU-positive myeloblasts in mice increased by 10.0% at 24 h af-ter intraperitoneal injection with HI E.coli,but decreased by 75.0% and 23.0% at 48 h and 72 h,respec-tively. The percentage of EdU-positive promyelocytes declined by 23.0%,54.5%,64.3% and 77.8% at 24 h,48 h,72 h and 96 h,respectively. The percentage of EdU-positive myelocytes increased by 60.0% and 10.0% at 24 h and 48 h,but decreased by 80.0% and 90.0% at 72 h and 96 h. The percentage of EdU-positive metamyelocytes increased by 50.0% at 24 h,but decreased by 33.3%,61.5% and 66.7% at 48 h,72 h and 96 h. The percentage of EdU-positive band and segmented cells increased by 14.0% at 24 h,but decreased by 50.0%, 77.8% and 88.0% at 48 h, 72 h and 96 h. Conclusion Emergency granulopoiesis occurred 24 h after the establishment of HI E.coli-induced model of acute peritonitis, which meant that the proliferation of myeloid precursor cells,especially that of myelocytes and metamyelocytes,was accelerated and resulted in increasing number of mature neutrophils immigrating to sites of inflammation.

Objective To investigate the effect of blood transfusion physicians for participating in clinical consultation to provide a reference for blood transfusion physicians participating in clinical practice.Methods The clinical data in 5 224 inpatients receiving blood transfusion before the blood transfusion physicians participating in clinical consultation (from January 2010 to December 2015) and after participating in clinical consultation (from January 2013 to December 2015) were retrospectively analyzed.Results The occurrence rates of blood transfusion adverse reactions before and after the blood transfusion physicians participating in clinical consultation were 2.39 ％ and 1.48 ％ respectively (x2 =5.674,P=0.021),the blood transfusion ratios of inpatients were 4.85 ％ and 3.55 ％ respectively (x2 =135.5,P＜0.01),the average allogeneic blood transfusion volumes were 0.563 U and 0.420 U (t=3.986,P=0.016),and the constituent ratios of mild,moderate andsevere anemia were 5.40％ and 0.23 ％,29.60％ and 17.90％,and 65.00％ and 81.80％ respectively (x2 =237.6,P＜0.01).Conclusion Participating in clinical consultation by the blood transfusion physicians is conducive to clinical treatment.

Objective: To further explore TCE-induced hepatotoxicity and its mechanisms by identification of trichloroethylene (TCE) induced abnormal histone methylation in human liver cells. Methods: L-02 cells were treated with 0 and 8 mmol/L TCE for 24 h. Histones were extracted by acid. Liquid chromatography electrospray ionization tandem mass spectrometry (ESI-LC-MS/MS) were used to identify and quantify TCE related histone methylations. TCE induced abnormal methylation of H3K79 me2 and H3K79 me3 were validated by Western blot analysis. The further analysis of the function of histone abnormal methylation modifications were done by single cell gel electrophoresis (SCGE) and Western blot analysis of p53 and ɤH2AX. Results: After treatment with TCE for 24 h in L-02 cells, the 36 TCE related histone methylation sites in 28 peptide segments were identified by MS. After treatment with TCE in concentrations of 0 and 8.0 mmol/L in L-02 cells for 24 h, the relative expression level of histone H3K79 me3 were 1.00±0.06, 0.70±0.09 (t=15.01, P=0.015); the relative expression level of histone H3K79 me2 were 1.00±0.05, 0.74±0.07 (t=16.69, P=0.018); the Olive Tail Moment about DNA damage were 1.46±0.28, 3.12± 0.68 (t=15.22, P=0.018); the relative expression levels of p53 were 1.00±0.04, 1.24±0.04 (t=18.71, P= 0.012); and the relative expression levels of ɤH2AX were 1.00 ± 0.03, 1.56 ± 0.11 (t=8.32, P=0 045). Conclusion TCE can induce changes in the relative expression level of H3K79 me2 and H3K79 me3 in L-02 cell, and induce DNA damage, suggesting that TCE may induce changes in the relative expression level of H3K79 me2 and H3K79 me3 by DNA damage.

Objective To evaluate the efficacy and toxicity of the combination of oxaliplatin and S-1 in the treatment of patients with advanced breast cancer.Methods A total of 72 patients with advanced breast cancer after the treatment failuer of anthracycline and taxane were treated with oxaliplatin and S-1.The first day,they were given oxaliplatin,135 mg/m2,with the 5％ glucose injection 500 ml,the time of intravenous drip should be more than 2 hours.And the S-1 was taken after breakfast and dinner,the dose was 40-60 mg,and the time of duration was 2 weeks,then they had 7 days to rest.The cycle was 21 days.Every 2 cycles,we estimated the efficacy.Patients who were effective and stable kept that chemotherapy regimens,the maximum duration was 6 cycles.The efficacy and toxicities were evaluated after cycles of chemotherapy.Results Two cases (2.8％) had complete response (CR),26 cases (36.1％) had partial response (PR).The response rate (RR) was 38.9％ and the disease control rate (DCR) was 69.4％.The median progress free survival (PFS) was 7.7 months and the median overall survival (OS) was 12.3 months.Subgroup analysis showed that the OS of patients who belong to stage Ⅳ,had two or more metastases or with failure treatment after being treated with anthracycline and taxane was notably shorter than the patients who belong to stage Ⅲ C,only one metastasis,with effective treatment after being treated with anthracycline and taxane,and the differences were statistically significant (10.5 months vs.15.0 months,x2 =4.469,P =0.035;9.3 months vs.15.0 months,x2 =8.297,P=0.004;10.0 months vs.14.0 months,x2 =4.077,P=0.043).The main side effects were neutropenia (19.4％),nausea (8.3％) and nerve toxicity (2.8％),mainly 3-4 degree,and could be welltolerated.The others were diarrhea,impaired liver function,stomatitis,anemia and hand-foot syndrome,mainly 1-2 degree.Conclusion Oxaliplatin combined with S-1 is effective and tolerable in treatment of patients with advanced breast cancer,the adverse reactions can be tolerated.

OBJECTIVETo assess the association of interferon gamma gene (IFNγ ) tag single nucleotide polymorphisms (Tag SNPs) with hepatitis B virus (HBV) infection in ethnic Dai and Hani minorities from Xishuangbanna, Yunnan.

METHODSPeripheral blood samples were collected from 300 Dai minorities and 300 Hani minorities, each included 100 healthy controls and 200 HBV infected individuals (including 100 spontaneous recovery subjects and 100 chronic HBV infected patients). Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDITOF-MS) was used to determine the Tag SNPs of IFNγ gene. Haplotypes were constructed.

RESULTSIn Hani and Dai minorities, the frequencies of rs1861494 CC genotype in HBV infected group was significantly higher than the healthy group (Dai: χ2=10.017, P=0.001; Hani: χ2=6.515, P=0.039), and there was a significant difference between the HBV infected group and the control group under the C allele recessive mode (CC/TC+TT) (Dai: P=0.035, OR=9.567, 95%CI: 1.166-78.499; Hani: P=0.027, OR=5.484, 95%CI: 1.216-24.726). In Dai minorities, the frequencies of rs2069705 CC genotype and C allele in chronic HBV infected group was significantly higher than the spontaneous recovery group (genotype: χ2=8.112, P=0.017; allele: χ2=4.066, P=0.044), and there was a significant difference between chronic HBV infected group and spontaneous recovery group under the C allele recessive mode (CC/CT+TT) (P=0.013, OR=0.341, 95%CI: 0.146-0.796).

CONCLUSIONAbove results suggested that the rs1861494 CC genotype of the IFNγ gene has conferred an increased risk for HBV susceptibility in both Dai and Hani minorities. In addition, the rs2069705 CC genotype may be a risky factor for Dai minorities to develop chronic HBV infection.

Adult ; Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Female ; Genetic Predisposition to Disease ; ethnology ; Genotype ; Hepatitis B ; ethnology ; genetics ; virology ; Hepatitis B virus ; physiology ; Humans ; Interferon-gamma ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Risk Factors ; Young Adult