BACKGROUND: Chinese medicines have better effects on preventing and treating diabetic nephropathy at early period, and the effects are caused by the diversity of the effective components of Chinese medicines which acts on the different targets of diabetic nephropathy.OBJECTIVE: To observe the effect of Xiaokening on the proliferation of mesangial cells under high glucose, and investigate the possible mechanism. DESIGN: A controlled observation.SETTING: Department of Endocrinology, the First Affiliated Hospital of Nanjing Medical University.MATERIALS: The experiments were completed in the Research Room of Cell Culture, Research Center of Endocrine Metabolism, the First Affiliated Hospital of Nanjing Medical University from July 2003 to May 2004. The rat mesangial cell lines HBZY-1 were bought from Chinese Center for Typical Culture Collection.METHODS: The mesangial cells were passaged and cultured according to the instructions. ① Grouping according to different concentration of stimulation: In the high glucose+Xiaokening group, Xiaokening of 6 concentrations were used, I.e., 20.0, 40.0, 60.0, 80.0, 120.0, 200.0 g/L. Meanwhile,normal glucose control group and high glucose control group were also set,the glucose concentration in the medium was 5.6 and 30 mmol/L respectively. The proliferations were observed after 72 hours. Grouping according to different time points of stimulation: There were normal glucose control group, high glucose control group and high glucose+Xiaokening group, the glucose concentration in the medium was 5.6, 30 and 30 mmol/L respectively, and the Xiaokening concentration was 60 g/L. The proliferations were observed at 24, 48 and 72 hours respectively in the three groups.② The dispensed cell suspension was placed into the 96-well plate, and 200 μL suspension for each well. The culture plate was placed in the inoculation box containing CO2 (0.05 in volume fraction) at 37 ℃ for 24 hours, then the supernatant was deserted, cell solutions containing different drugs of different concentrations were added in each group, 200 μL for each well. The 96-well plate was then placed in the culture box containing CO2 (0.05 in volume fraction) and 100% humidity at 37 ℃ for inoculation.In the groups of different concentrations, 20 μL MTT (5 g/L) was added into the wells after 72 hours. In the groups of different time points, 20 μL MTT (5 g/L) was added into the wells at 24, 48 and 72 hours. Then the plates were inoculated at 37 ℃ for 4 hours. Under microscope, it was observed that MTT get into the cells, the supernatant was sucked away, then 150 μL dimathyl sulfoxide was added to dissolve methyl alcohol, and shaken to even with plate rocking bed for 30 s, and the absorbance (A) values were recorded with the microplate reader at the wavelength of 492 nm.MAIN OUTCOME MEASURES: The proliferations of mesangial cells (A value) were observed at different time points and in Xiaokening of different concentrations.RESULTS: ①Proliferation of mesangial cells at different time points: The A values in the high glucose control group at 24, 48 and 72 hours (0.685 ±0.010, 0.750±0.087, 0.659±0.018) were higher than those in the normal glucose control group (0.586±0.054, 0.598±0.040, 0.527±0.047, P ＜ 0.05). In both the high and normal glucose control groups, the A value at 48 hours was higher than that at 24 hours (P ＜ 0.05), but the A value at 72 hours was lower than that at 48 hours (P ＜ 0.05). The A value in the high glucose+Xiaokening group at 72 hours was 0.538±0.023, and it was lower than that in the high glucose control group (P ＜ 0.05). ② Proliferation of mesangial cells in Xiaokening of different concentrations: Xiaokening high er than 60.0 g/L could obviously restrain the excessive stimulation of high glucose to the mesangial cells, and the effect was in a concentration-de pendent manner, but too high concentration (120.0 g/L) would result in the abscission and death of cells, whereas no survived cells could be observed in extremely high concentration (200.0 g/L). CONCLUSION: Xiaokening could inhibit the proliferation of mesan gial cells stimulated by high glucose after 72 hours in a concentration dependent manner, but too high concentration would cause strong cytotoxicity.

Objective To investigate the value of real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) in the detection of Streptococcus agalactiae in early screening of Streptococcus agalactiae in pregnant women in the third trimester, and to study the effect of Streptococcus agalactiae infection on them.Methods A retrospective analysis of 5 855pregnant women with vaginal discharge and rectal secretions from January 2017to January 2018in our hospital were performed to detect Streptococcus agalactiae by RT-qPCR.Meanwhile, 100vaginal non-vaginosis women in the third trimester were selected as the negative group and100healthy women as the control group.The vaginal micro-environment-related indexes (Lactobacilli content, flora diversity and cleanliness) of three groups were analyzed.Results The total carrier rate of Streptococcus agalactiae of the third trimester from January 2017to January 2018in our hospital was 9.6％, of which vaginal carriage rate was 2.4％, rectum carriage rate was 7.2％, Streptococcus agalactiae positive group (GradeⅡ-Ⅲ), rates of population diversity (GradeⅡ-Ⅲ) cleanliness (Ⅰdegree) and microenvironment imbalance were 30.1％, 36.4％, 25.9％and 76.2％respectively, while those in the negative group were 58.0％, 55.0％, 61.0％and 63.0％and those in the control group were 87.0％, 91.0％, 83.0％, 24.0％.The positive rate of Streptococcus agalactiae in each group was significantly higher than that in the control group (P＜0.05).The differences of all the indexes between the negative group and the control group were also statistically significant (P＜0.05).Conclusion The carrying rate of Streptococcus agalactiae in third trimester pregnancy in our hospital was within the normal range reported in the literature.The detection of Streptococcus agalactiae by RT-qPCR could be effectively screened and monitored.Carrying Streptococcus agalactiae might increase the risk of vaginal infection in third trimester women.