Objective Establish the method of bacterial endotoxins test for human papillomavirus bivalent(types 16,18)vaccine(pichia pastoris).Methods The method of gel-clot technique and photometric technique(kinetic-turbidimetric assay and kinetic-chromogenic assay)for the determination of bacterial endotoxins was carried out according to the Appendix of China Pharmacopoeia in 2020 Vol Ⅲ 1143"bacterial endotoxins test"and Vol IV 9251"guidelines for bacterial endotoxin test".Results There is no interference after more than forty times dilution for gel-clot technique,the recovery rates of kinetic-turbidimetric assay and kinetic-chromogenic assay can also meet the requirements.Conclusion It is feasible to establish gel-clot technique,kinetic-turbidimetric assay and kinetic-chromogenic assay for bacterial endotoxin test of human papillomavirus bivalent(types 16,18)vaccine(pichia pastoris).

Objective:To investigate the effect of dydrogesterone (DYD) on the ultrastructure of the endometrium in the window of implantation.Methods:We prospectively studied endometrium tissues of patients who underwent in vitro fertilization and embryo transfer (IVF-ET) from July 2020 to July 2022 at the Center for Reproductive Medicine of Zhejiang Provincial People's Hospital. The ultrastructure of mid-luteal endometrium was examined by scanning electron microscopy and transmission electron microscopy. Firstly, we used self-control study to compare the ultrastructure of endometrium between the natural cycle group and the DYD 40 mg/d group in 20 patients with male factor infertility. Secondly, we used self-control study to compare the ultrastructure of the endometrium between the DYD 40 mg/d group and the DYD 80 mg/d group in 43 patients with recurrent implantation failure (RIF). Results:There were no significant differences in endometrial thickness, ultrasound grading, positivity rate of nucleolar channel system (NCS), pinopode score and secretor status score, in addition to number and morphology score of mitochondria, between the natural cycle group and the DYD 40 mg/d group (all P>0.05). In RIF patients, the DYD 80 mg/d group rendered significantly higher pinopode score (-46.26±19.97), NCS positivity rate [65.12% (28/43)] and mitochondrial morphology score [0 (0, 1)] than those in the DYD 40 mg/d group [-67.62±15.94, P<0.001; 39.53% (17/43), P=0.043; 0 (0, 0), P=0.023], while other characteristics were not significantly different between the two groups (all P>0.05). Conclusion:The ultrastructure of endometrium in DYD supplementation cycle was similar to that in the natural cycle. Increasing the dosage of DYD significantly improved pinopode maturity, NCS positivity rate and mitochondrial morphology of the endometrium in RIF patients.

Objective:To investigate the effect of dydrogesterone (DYD) on the ultrastructure of the endometrium in the window of implantation.Methods:We prospectively studied endometrium tissues of patients who underwent in vitro fertilization and embryo transfer (IVF-ET) from July 2020 to July 2022 at the Center for Reproductive Medicine of Zhejiang Provincial People's Hospital. The ultrastructure of mid-luteal endometrium was examined by scanning electron microscopy and transmission electron microscopy. Firstly, we used self-control study to compare the ultrastructure of endometrium between the natural cycle group and the DYD 40 mg/d group in 20 patients with male factor infertility. Secondly, we used self-control study to compare the ultrastructure of the endometrium between the DYD 40 mg/d group and the DYD 80 mg/d group in 43 patients with recurrent implantation failure (RIF). Results:There were no significant differences in endometrial thickness, ultrasound grading, positivity rate of nucleolar channel system (NCS), pinopode score and secretor status score, in addition to number and morphology score of mitochondria, between the natural cycle group and the DYD 40 mg/d group (all P>0.05). In RIF patients, the DYD 80 mg/d group rendered significantly higher pinopode score (-46.26±19.97), NCS positivity rate [65.12% (28/43)] and mitochondrial morphology score [0 (0, 1)] than those in the DYD 40 mg/d group [-67.62±15.94, P<0.001; 39.53% (17/43), P=0.043; 0 (0, 0), P=0.023], while other characteristics were not significantly different between the two groups (all P>0.05). Conclusion:The ultrastructure of endometrium in DYD supplementation cycle was similar to that in the natural cycle. Increasing the dosage of DYD significantly improved pinopode maturity, NCS positivity rate and mitochondrial morphology of the endometrium in RIF patients.

Objective:To analyze the evaluation value of changes in β-human chorionic gonadotrophin (β-HCG) for the curative effect of methotrexate (MTX) on ectopic pregnancy (EP).Methods:A retrospective analysis was performed in 80 EP patients who admitted to Jinhua People's Hospital from January 2013 to August 2019.Results:Of the 80 EP patients, success and failure rates of treatment were 77.50% (62/80) and 22.5% (18/80), respectively.According to different treatment results, they were divided into success group (62 cases) and failure group (18 cases). Before treatment, hemodynamic index of fallopian tube artery[resistance index (RI)] in success group was higher than that in failure group ( t=4.150, P<0.05). Before treatment, after 1d, 4d and 7d of treatment, the serum levels of β-HCG in success group were lower than those in failure group ( t=69.496, 76.618, 217.488, 310.203, all P<0.05). The results of Pearson correlation analysis showed that RI was negatively correlated with serum level of β-HCG before treatment, after 1d, 4d and 7d of treatment ( r=-0.374, -0.396, -0.471, -0.482, all P<0.05). The receiver operating characteristic (ROC) curves analysis showed that after 4d and 7d of treatment, areas under the curve (AUC) of β-HCG for evaluating the curative effect of MTX on EP were 0.821 and 0.829, respectively, which were higher than before treatment and after 1d of treatment (0.685, 0.701) ( P<0.05). Conclusion:The changes of β-HCG are of high clinical value in evaluating the curative effect of MTX on EP, which can be applied as an important monitoring index.

Objective It is important to precisely determinate the single nucleotide polymorphisms (SNPs) in many genes in‐cluding genes related with base excision repair (BER) pathway .This research is conducted to evaluate the role of polymerase chain reaction with confronting two‐pair primers (PCR‐CTPP) in analyzing the SNPs of BER pathway .Methods Four common SNPs of BER pathway (OGG1 Ser326Cys ,XRCC1 Arg399Gln ,APE1 Asp148Glu and‐141T/G in the promoter region) was detected with PCR‐CTPP .10 of the products were sent for genotype sequencing .Compare the results of PCR and sequencing to evaluate the accu‐racy of PCR‐CTPP .Results The genotypes were exactly the same as the sequencing .Conclusion The PCR‐CTPP was a reliable and rapid detective technology for SNPs genotyping .Its broadest application would be great help for gene variant analysis .

Objective To investigate the present state of mineral metabolism and its related factors in elderly patients with maintenance hemodialysis (MHD),in order to improve the quality of life and survival rate in elderly patients with MHD in Xinxiang.Methods Clinical data of 466 patients with treatment of MHD for over 3 months in 4 hospitals in Xinxiang district were enrolled from January 2012 to August 2013 in this study,and the patients were divided into the elderly group (n=159,aged ≥ 60 years) and the non-elderly group (n =307,aged ＜ 60 years).The evaluation indexes for mineral metabolism were determinated,such as serum levels of calcium,phosphorus,intact parathyroid hormone (iPTH) and alkaline phosphatase (ALP).The present state of mineral metabolism and its relationships with anemia,dialysis ages,adequacy of dialysis,malnutrition and inflammatory factors were analyzed in elderly patients with MHD.Results Among 466 patients,the serum levels of calcium,phosphorus and iPTH in MHD patients were (1.95 ± 0.34) mmol/L,(2.54±1.38) mmol/L and (409.5±345.6) ng/L respectively,and their control rates were 34.3％,20.4％ and 25.5％ respectively.And in patients with hemodialysis three times a week,the control rates were 50.8％,31.4％ and 32.2％ respectively.The serum levels of phosphorus,iPTH,ALP,albumin and transferrin saturation were lower in elderly group than in non-elderly group (all P＜0.01),and the serum levels of pre-albumin,hemoglobin and feritin were lower in elderly group than in non-elderly group (all P＜0.05).But the serum levels of calcium and C-reactive protein (CRP)were higher in elderly group than in non-elderly group (all P＜0.05).Conclusions The elderly patients with MHD exhibit unique clinical manifestations of abnormal mineral metabolism,and low bone transformation osteopathy is the main manifestation.The micro-inflammation,malnutrition and anemia are more severe in elderly patients than in non elderly patients.

Objective To analyze the expression characteristics of chemotherapeutic related genes (ERCC1 ,BRCA1 ,APE1 , RRM1 ,TUBB3 and TS) in non-small cell lung cancer(NSCLC) and their associations with the pathological types to provide the ex-perimental evidences for clinical individual chemotherapy .Methods The immunohistochemical SP method was used to determine the expression of ERCC1 ,BRCA1 ,APE1 ,RRM1 ,TUBB3 and TS genes in 733 patients with NSCLC .Chi-square test was performed to analyze the gene expression characteristics among different pathological types .Spearman relationship was conducted to evaluate the associations among the six genes and different pathological types .Results The expression of BRCA1 ,TUBB3 and TS had sta-tistical differences among three pathological types (P<0 .01);and these three genes showed a relationship with the pathological type (r=0 .107 ,-0 .229 and 0 .168 ,respectively ,P<0 .01) .ERCC1 vs .BRCA1 ,ERCC1 vs .APE1 and APE1 vs .RRM1 showed the pos-itive correlation(r=0 .214 ,0 .316 and 0 .222 ,respectively ,P<0 .01) .Conclusion The chemotherapeutic related gene expressions of NSCLC are related with the clinical pathological types .And the expressions of multiple markers are associated with each other .The combined detection of the chemotherapeutic related markers and the pathological types would be helpful to the decision making of individual chemotherapy in clinic .

OBJECTIVE:To optimize the technics for the extraction of Qiangliyanhou mixture.METHODS:The physico-chemical properties of the compositions in the formula medicinal substances were analyzed,the obtained quantity of aqua aromatica was used as an investigation index of distillation technics;The preparation technics was optimized by orthogonal experiment in which the extraction ratio of the water extractives and the content of chlorogenic acid were used as the indexes to investigate the influence of the amount of water added,the soaking time before decoction,decoction time and times of decoc?tion on the extraction result.RESULTS:The optimized amount of aqua aromatica in the distillation procedures was collecting distillate that is3times the amount of the medicinal substances;The optimum water extraction condition was:the medicinal substances was soaked for2hours by adding15times amount of water before decocting for total3hours(3times?1hour/time).CONCLUSION:The preparation procedure is reasonable and reliable.