Purpose: To investigate the clinical efficacy of blood-letting puncture and cupping in cooperation with acupuncture and moxibustion for treating Ramsay Hunt syndrome. Atreatment group of 32 cases was treated by blood-letting puncture with plum-blossom needle and cupping in cooperation with acupuncture and moxibustion and a control group of 32 cases,with acupuncture and moxibustion, and andaciclovir. Results and conclusion: The cure and marked effectiveness rate was 81.2% in the treatment group and 56.2% in the control group after 4 courses of treatment. It was higher in the treatment group than in the control group (P<0.05). The short-term otalgia-relieving rate was also significantly higher in the treatment group than in the control group (P<0.01).

Objective To construct the RNA interference eukaryotic expression vector targeting human centromere protein-I (CENP-I) and to observe its effect on the growth of human embryo kidney 293 cells (HEK 293). Methods The expression vectors of pGenesil-1/CENP-I-siRNA-1. pGenesil-1/CENP-I-siRNA-2 and pGenesil-1/CENP-I-siRNA-3 were constructed by gene recombination and then were transfected into the HEK293 cells by liposome. The expressions of CENP-I at the protein and mRNA levels were detected by Western blotting and fluorescence quality PCR (FQ-PCR). The effective vector and the best transfection time were selected. The growth and the cell cycle of the transfected cells were assessed by MTT assay and flow cytometry. Giemsa was used to stain the transfected cells to calculate the mitotic index. Results Sequence-specific siRNAs targeting CENP-I significantly down-regulated the expression of CENP-I in HEK293 cells. The recombinant plasmid of pGenesil-1/CENP-I-siRNA-3 was the effective vector. After transfecting for 72 h the best inhibited efficiency was achieved. In CENP-I-siRNA transfected cells,the rate of cell growth was decreased markedly. Cells at G 2/M phase and the mitotic index were increased conspicuous compared with the cells transfected with the blank vector or untransfected. Conclusion Down-regulation of CENP-I in HEK293 cells by sequence specific siRNA delays the cell growth and postpones the cell division.

Objective To compare the cost-effectiveness and cost-utility of venlafaxine and mirtazapine in patients with treatment-resistant major depression (TRD).Methods One hundred and five patients with TRD were enrolled in this study and grouped into venlafaxine treatment (n=50) and mirtazapine treatment (n=55) based on the double-blind randomization scheme generated by computer.The treatment costs of antidepressants during 8 weeks were calculated,the rates of clinical response and remission were taken as treatment effectiveness,and the quality-adjusted life years (QALYs) as treatment utility.The descriptive analysis and nonparametric test were used to compare the cost-effectiveness and cost-utility of different groups.Results During 8 weeks,the treatment cost of antidepressant was ￥ 1 396.44 for venlafaxine and ￥ 1 206.90 mirtazapine,and the difference between two groups was ￥ 189.54.The cost-effectiveness ratios between venlafaxine and mirtazapine were very close (differed ￥ 0.06 for remission rate and ￥ 1.08 for response rate respectively).There was no significant difference for cost-utility ratios between two groups (physical functioning Z=-0.15,P＞0.05 ; mental health Z=-0.54,P＞0.05).Conclusion Both cost-effectiveness and cost-utility of venlafaxine in patients with TRD are close between venlafaxine and mirtazapine.

Objective To investigate the expression of human spindle mitosis arrest deficiency gene (hsMAD2)in spontaneous abortion embryos and the relationship between low expression of hsMAD2 and numerical chromosomal aberration. Methods Spontaneous abortion embryo tissues were collected,including 23 cases of once spontaneous abortion tissue and 10 cases of twice or more spontaneous abortion tissue and induced abortion embryos(35 cases)from the Department of Gynaecology and Obstetrics of the Affilisted Hospitals of Chongqing University of Medical Science during the period of March 2006 to March 2007.FQ-PCR and western blot were used to evaluate the endogenous expression level of hsMAD2 mRNA and hsMAD2 protein;primary culturing of cells from the induced abortion embryos was conducted and 5 embryonic cells were selected by chromosomes karyotype analysis.Recombinant shRNA plasmids targeting hsMAD2 gene were constructed to inhibit the expression of endogenous hsMAIY2 genes in embryonic cells which have normal karyotypes;the groups were defined as the first experimental group(transfeeted with pshRNA-hsMAD2-1),the second experimental group(transfected with pshRNA-hsMAD2-2),the third experimental group(transfected with pshRNA-hsMAD2-3),the first control group(transfected with nothing),the second control group(transfected with pTZU6+1)and the independent group(transfected with pshRNA-N1).Interference efficiency was demonstrated by FQ-PCR and western blot:cell prolireration was meagured by methyl thiazolyl tetrazolium(MTT)assay;cell-cycle was assessed by flow cytometry (FCM):the chromosome numbers were calculated to analyze the variation of chromosomes.Results(1) The mRNA levels of hsMAD2 in the once spontaneous abortion tissue,twice or more spontaneous abortion tissue and indueed abortion tissue were 0.00879±0.00035.0.00901±0.00033 and 0.00941±0.00026 respectively,and there Wag no significant ditierence(P＞0.05)compared with each other;however,the protein levels of hsMAD2 in three groups were 0.2791±0.0311.0.0431±0.0020 and 0.5790±0.0331 respectively,and there were significant difierences(P＜0.05)compared with each other.(2)Recombinant shRNA plagmids could significantly and specifically inhibit hsMAD2 gene expression in embryonic cells.Compared with the first control group(4%)and the second control group(3%),the recombinant shRNA could inhibit embryonic cell proliferation to 54% at 48 h after transfection(P＜0.05):compared with the first control group(8.2%)and the second control group(8.0%),the ratios of G2/M phase cells in the experimental group(17.9%)was significantly increased(P＜0.05);compared with the first control group (4.8%),the ratios of abnormal chromosomes in the experimental group was increased to 30.0%(P＜0.05).Conclusions Down-expression of hsMAD2 gene may be one of the mechanisms inducing numerical chromosome aberration,abnormal embryo development and the occurrence of spontaneous abortion.

Objective To explore the mechanism of hBUB1 gene in the developing of spontaneous abortion embryos with numerical chromosomal abnormalitv.Methods Quantitative real-time RT-PCR and Western blot were used to determine the mRNA and protein level of hsMAD2 gene both in spontaneous abortion embryos with numerical chromosomal abnormality(experimental group)and with numerical chromosomal normality(control group).Recombinant shRNA plasmids targeting hBUB1 gene was constructed to inhibit the expression of endogenous hBUB1 genes in embryonic cells.Interference efficiency was demonstrated by fluorescent quantitative PCR and Western blot.The inhibitory rate of cell proliferation was measured by MTT assay and cells-cycle was assessed by flow cytometry.Resuns Western blot analysis showed that protein level of hBUB1 in the experimental group was decreased signifieandv(the rate of positivity and strong positivity were 8%and 93.5%,respectively,P＜0.05)compared with the control group.The expression of hBUB1 gene in embryonic cells was significantly and specially inhibited by shRNA plasmids (the mRNA level before and after treatment witll RNAi were 0.196±0.067 and 0.042±0.006,respectively,P＜0.05).The inhibitory rate of cell proliferation was increased to 62%from 4%at 48 h after transfection.The rate of G2/M phase cells was decreased after transfection with efficient shRNA(control group:40.2%and 41.3%,test group:21.3%).Conclusions Down-regulation of hBUB1 gene leads to the inhibition of cell proliferation and the arrest of cell-cycle.It also probably plays an important role in the development of spontaneous abortion embryos with numerical chromosomal abnormality.The clinical relevance warrant further investigation.

Purpose:To study bcl-2 and MDR expressions in renal cell carcinoma and to evaluate its clinical significance.Methods:blc-2 and MDR expression were studied by immunohistochemical technique in 48 specimens of renal cell carcinoma tissues and in 10 normal renal tissues and their clinical significance was analyzed.Results:The positive rate of expression of bcl-2 in the renal cell carcinoma tissue was 77.1%, while it was 40.0% in the normal renal tissue(P＜0.05). However, it was not related to the tumor grade and tumor stage(P＞0.05). On the other hand, There was a positive expression of MDR1 in renal cell carcinoma and normal renal tissue. MDR1 was positively related to tumor cell type(P＜0.05). Compared with the other types of renal cell carcinoma. The granular existed higher expression. Both have no relationship with prognosis.Conclusions: bcl-2 Protein is overexpressed in the majority of renal cell carcinomas. The overexpression of bcl-2 may have a role in tumorigenesis. MDR1, an established predictor for chemo-resistance, however, has no significant correlation with bcl-1. The multidrug resistant of renal cell cancer is probably induced by double-functions of bcl and MDR1.

Objective To systematically evaluate the correlation between vitamin D deficiency in early pregnancy and the outcome of preterm birth.Method PubMed,Embase,the Cochrane Library,Web of Science,Ebsco,CBM,CNKI and Wanfang Data databases were searched to collect cohort studies and case-control studies on the correlation between vitamin D deficiency in early pregnancy and preterm birth outcomes,and the retrieval time was from the establishment of the database to June 2019.Two researchers independently reviewed the literature,extracted the data and evaluated the risk of bias in the included studies.RevMan 5.3 software was used for Meta analysis.Result A total of 6 cohort studies and 3 nested case-control studies were included.A total of 30 891 newborns were included,including 1 912 premature infants.3 Chinese articles and 6 English articles were reviewed including three studies from China,three from North America,two from Europe and one from Australia.The diagnostic criteria for vitamin D deficiency and preterm birth were similar in these studies.After adjusting for age,race and other confounding factors,Meta-analysis results showed that vitamin D deficiency in early pregnancy did not increase the risk of preterm birth (OR =1.04,95％ CI 0.90 ～ 1.20,P =0.63).Subgroup analysis were conducted according to the study type,measurement method and regional population,and the results were consistent with the overall results.No significant publication bias was found in the meta-analysis results.Conclusion Current evidence suggests that vitamin D deficiency in early pregnancy has no significant influence on preterm birth.

Objective To investigate the effects of different culture methods in vitro mouse zona-free embryonic devel-opment. Methods In various stages of embryonic development rate,blastocyst rate and cell numbers of blastocysts de-veloped were measured as mWOW system (the modified “well of well” system), micro-drops single + group, micro-drops single and micro-drops group culture in vitro. The effects of different methods of were compared in vitro embryo development zona-free. Results Embryonic development rate,blastocyst rate and cell numbers of blastocysts developed were significantly higher in mWOW system culture than micro-drops single + group, micro-drops single and micro-drops group culture. Conclusion Compared with micro-drops single+group,micro-drops single and micro-drops group culture,mWOW system method is more suitable for zona-free cultured in vitro mouse embryos.

OBJECTIVETo investigate the effect of high-intensity focused ultrasound (HIFU) on tumor metastasis in mouse model bearing melanoma xenograft.

METHODSMice bearing murine melanoma B16-F10 cell xenograft were randomized for sham-HIFU or HIFU exposure when the tumors grew to a maximum diameter of 7-10 mm, and the tumor size was measured every 3 days. The cumulative survival rate of the mice and tumor metastasis rate were calculated, and the circulating melanoma cells were detected using qRT-PCR. At 14 days after HIFU treatment, B16-F10 cells were retransplanted via the tail vein and the pulmonary metastatic nodules were counted.

RESULTSThe median survival time of the mice was 19.00 days (95% CI 17.14-20.86 days) in the sham group and 26.00 days (95%CI 24.76-27.25 days) in HIFU group. The cumulative survival rate in the HIFU group was significantly higher than that in sham-HIFU group (P<0.01), and the tumor size was significantly smaller in HIFU group at 20, 23, and 26 days after HIFU treatment (P<0.05). Compared with the sham-HIFU group, HIFU group had significantly lower levels of MAGE-A3, MART1 and PAX3 at 7 days after HIFU (P<0.05) with still lower MAGE-A3 level at 14 days (P<0.05). HIFU group showed a significantly smaller number of pulmonary metastatic nodules following tumor cell retransplantation than in sham-HIFU group (P<0.01) with a metastasis inhibition rate of 42.4%.

CONCLUSIONHIFU treatment can inhibit tumor metastasis in melanoma-bearing mice possibly by reducing tumor cell detachment from the primary tumor site and suppressing colonization of the circulating melanoma cells.

Objective To investigate the effect of high-intensity focused ultrasound (HIFU) on tumor metastasis in mouse model bearing melanoma xenograft. Methods Mice bearing murine melanoma B16-F10 cell xenograft were randomized for sham-HIFU or HIFU exposure when the tumors grew to a maximum diameter of 7-10 mm, and the tumor size was measured every 3 days. The cumulative survival rate of the mice and tumor metastasis rate were calculated, and the circulating melanoma cells were detected using qRT-PCR. At 14 days after HIFU treatment, B16-F10 cells were retransplanted via the tail vein and the pulmonary metastatic nodules were counted. Results The median survival time of the mice was 19.00 days (95 % CI 17.14-20.86 days) in the sham group and 26.00 days (95%CI 24.76-27.25 days) in HIFU group. The cumulative survival rate in the HIFU group was significantly higher than that in sham-HIFU group (P<0.01), and the tumor size was significantly smaller in HIFU group at 20, 23, and 26 days after HIFU treatment (P<0.05). Compared with the sham-HIFU group, HIFU group had significantly lower levels of MAGE-A3, MART1 and PAX3 at 7 days after HIFU (P<0.05) with still lower MAGE-A3 level at 14 days (P<0.05). HIFU group showed a significantly smaller number of pulmonary metastatic nodules following tumor cell retransplantation than in sham-HIFU group (P<0.01) with a metastasis inhibition rate of 42.4%. Conclusion HIFU treatment can inhibit tumor metastasis in melanoma-bearing mice possibly by reducing tumor cell detachment from the primary tumor site and suppressing colonization of the circulating melanoma cells.