OBJECTIVETo study the effect of total flavanones of Sedum sarmentosum (SSTF) on the apoptosis of rat hepatic stellate cells (HSC-T6) and its mechanism.

METHODDifferent concentrations of SSTF and HSC-T6 cells were co-cultured for different period of time. The MTT assay was used to detect the inhibitory effect of SSTF on the proliferation of HSC-T6 cells. The flow cytometry Annexin-V/PI double staining method was adopted to detect SSTF's effect on HSC-T6 cell apoptosis. Western blotting and Real-time PCR methods were applied to observe the effect on the protein and mRNA expressions of apoptosis-related cytokines Bcl-2, Bax and Caspase-3.

RESULTSSTF significantly inhibited HSC-T6 cell proliferation and induced cell apoptosis in a dose and time dependent manner. According to Western blotting result, SSTF promoted apoptosis by inhibiting Bcl-2, Bax and promoting the protein expression of Caspase-3; according to a further Real-time PCR study, Bcl-2 mRNA levels can inhibit Bcl-2 and promote Bax and Caspase-3 expressions.

CONCLUSIONSSTF has the effect of promoting the apoptosis of HSC-T6 mainly by inhibiting Bcl-2 and promoting protein and mRNA expressions of Bax and caspase-3.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Cell Line ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Flavanones ; pharmacology ; Hepatic Stellate Cells ; cytology ; drug effects ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Rats ; Sedum ; chemistry

Objective In order to obtain the reference sequences and relative expression of transcription genes and study the genetic base of active ingredients in Penthorum chinense, which were useful for researching functional gene to P. chinense. Methods In this study, by performing Illumina Hiseq 2000 and de novo assembly, the transcriptome of whole plant was sequenced, the data were filtered and assembled, and the unigene was compared and annotated. Meanwhile, the genes related to the synthesis of metabolic pathway of active ingredients in P. chinense were analyzed. Results Totally, 40 005 442 valid short sequences were obtained, and 42 306 unigenes were spliced by de novo. Also, a total of 518 open reading frames (ORF) were obtained by ORF analysis, and 75 ORF of them had transcription factor domains. In addition, by performing KEGG pathway analysis, 33, 32, 59, and 68 unigenes were found to be involved in the pathway of flavonoid biosynthesis, steroid biosynthesis, terpenoid backbone biosynthesis, and 2-oxocarboxylic acid metabolism, respectively. Conclusion The datasets provided in this study will contribute significantly to genetic improvement and study on the genes related to biosynthesis pathway of pharmaceutical active substances from P. chinense.

OBJECTIVE: To establish a one-step esterification derivative analysis method for fatty acids in Cordyceps sinensis. and analyze and compare the composition and content of fatty acids of 12 samples from five counties of Yushu prefecture of Qinghai province. METHODS: Qualitative and quantitative analysis of fatty acids was performed by gas chromatography-mass spectrometry (GC/MS) in combination with comparison to reference substances, then the data was compared. RESULTS: Thirteen kinds of fatty acids of C14-C20 were found with a total amount of 79.81 - 160.20 mg · g^-1. For a same sample, palmitic acid, oleic acid, and linoleic acid were the three kinds of fatty acids with the highest contents, the total content of which accounted for 92.81% -95.66% of the total 13 fatty acids. The proportion of unsaturated fatty acid was much higher than that of saturated fatty acids, and the ratio of UFA/SFA was 4.23-6.63. The fatty acids of 18 carbon and 16 carbon were the component with the highest amount and content, and the ratio of C18/C16 ranged from 4.04 to 6.19. Furthermore, the composition of the 13 fatty acids was basically consistent, while the content and proportion of the fatty acids varied greatly in samples from different origins. CONCLUSION: The method of fatty acids determination in Cordyceps sinensis set up by this research is simple, accurate, and reproducible, which is suitable for the fatty acids analysis in natural Cordyceps sinensis. The qualitative and quantitative data of the fatty acids could provide references for better recognition of the value of Cordyceps sinensis and further studies on its quality control.

OBJECTIVE: To establish an HPLC method for simultaneous determination of 15 nucleosides in Cordyceps sinensis, and investigate the differences in the composition and contents of these 15 nucleosides in 12 samples from 5 counties of Yushu prefecture, Qinghai province.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Interleukin-6 ; metabolism ; Liver Neoplasms ; metabolism ; pathology

Oral glucose tolerance test(OGTT)was performed in 419 first-degree relatives(FDRs)of type 1 diabetes mellitus. GADA, IA-2A, and IAA were determined by radioligand assay, and the positive rates were 7.16%, 1.43%, and 1.26%, respectively. Intravenous glucose tolerance test(IVGTT)and nateglinide-OGTT were performed in 39 controls, 11 first-degree relatives with positive autoantibody(Ab+group), 14 ones with negative autoantibody(Ab-group)during 5-7 days.The first-phase insulin release(FPIR), area under insulin release during 0-10 min [AUC0-10] of IVGTT and the value of(ΔI30/ΔG30)of nateglinide-OGTT in Ab+group were lower than those of control and(2.75±0.37 vs 3.61±1.05)mU/mmol, all P＜0.05]. The 1st min insulin release in Ab+group was lower than that of Ab-group [(3.80±0.30 vs 4.52±0.70)mU/L, P＜0.05]. The HOMA-IR was higher in Ab-group than that in control group(2.92±1.04 vs 1.96±1.22, P＜0.05). The results suggest that the positivity rates of autoantibodies in FDRs of type 1 diabetes mellitus are very close to those of Caucasian. There exist insulin secretion defects in FDRs with positive autoantibody while insulin resistance in FDRs with negative autoantibody.

Objective To investigate the expression of Oct4 in liver cancer, and the interrelation of the Oct4 and Wnt/β-catenin genes in hepatocellular carcinoma( HCC) cell line HepG2. Methods RTPCR technique was used to detect the expression of Oct4 and β-Catenin in HCC specimens; RNAi was used to knock-down the expression of Oct4 in HepG2, and the change of Wnt/β-catenin related genes were detected by Real time-PCR. Results In HCC specimens, the expression of Oct4 and β-Catenin in tumor and cirrhotic liver tissues were stronger than normal liver tissues. In SiRNA Oct4 HepG2 cells, the expression of Oct4 was downregulated, and β-catenin as well as Wnt10b were in a positive correlation with Oct4, TCF3 was in negative correlation with Oct4. Clone formation and move ability of the HepG2 were downregulated. Conclusions The expression of Oct4 was higher in tumor tissues than in normal liver tissues. Silencing Oct4 by SiRNA-0ct4 in HepG2 resulted in decreased ability of clone formation and cell movement.

Objective To find methods to isolate and purify plasminogen activator (Pla) from artificial culture of Yersinia pestis. Methods Ultrasonication and urea extracting combined by ammonium sulfate salting-out were tried to extract Pla. High performance liquid chromatography(HPLC) was used to purify Pla. The first step was ion exchange and the second was gel filtration, Preparative electrophoresis was used to purify Pla, too. The enzyme activity of the isolated or purificated Pla was detected. Results Both 50% - 60% saturated ammonium sulfate deposition of supernatant of plague bacilli ultrasonication and 0 - 10% saturated ammonium sulfate deposition of supernatant of plague bacilli powder soaked by urea had three bands(Mr about 31×103, 35×103 and 37×103) and lysis rings were 6.5 and 7.2 mm in diameter respectively when the enzyme activity was detected. Pla purified by HPLC was mainly composed of three bands(Mr about 31×103, 35×103 and 37×103), occupying more than 80% of total protein weight and lysis ring was 5.0 mm in diameter. Pla purified by preparative electrophoresis mainly consisted of three bands(Mr about 31×103, 35×103 and 37×103) with other proteins of low concentration nearby, no lysis ring was detected. Conclusions Pla is collected by the methods of ultrasonication and urea extracting. Priliminary purification of Pla can be achieved by HPLC and preparative electrophoresis.

Objective To investigate the clinical value of evaluation of fetal cardiac function in congenital heart disease by brain natriuretic peptide (BNP) and velocity vector imaging (VVI). Methods Fetuses who came from Shenzhen Maternity & Child healthcare Hospital were divided into the congenital heart disease group and the control group. At the same time we collected amniotic fluid and assayed BNP concentration. Using the VVI software, the velocity, strain and strain rate of the global and segmental of the left ventricle were measured. Comparison and correlation were made between the two groups. Results There was significantly difference of BNP concentrations in amniotic fluid between two groups. The gestational age had significant positive correlation with BNP concentrations in disease group. The comparison of global velocity, strain and strain rate of left ventricle between the two groups showed significant differences. All of the left ventricular dynamic parameters in disease group were lower than those of the control group. Conclusions Compared with the control group, the disease group had a high level of BNP in amniotic fluid and a lower level of dynamic parameters of left ventricular. There was a positive correlation between BNP concentration and gestational age in disease group. So we can conclude that theBNP concentration can be a biological parameter for evaluating the latent impairments of fetal cardiac function.

Objective To investigate the relationship between spontaneous miscarriage and embryonic chromosome abnormalities,and to evaluate the clinical application of karyotype analysis by chorionic villus cell culture. Methods The chorionic villus karyotype of 1983 cases of miscarriage from January 2010 to July 2016 in Guangzhou Women and Children′ s Mecical Center were analyzed retrospectively. The miscarried chorionic villi were obtained by curettage under sterilized condition. The chromosome specimens were prepared after chorionic villus cell culture. Karyotype analysis was performed by G-banding technique. Results In the 1983 samples, successful karyotype analysis was performed in 1770 cases, with the successful rate of 89.98%. Chromosomal abnormalities were found in 1038 cases (58.64%,1038/1770). Chromosomal structural abnormalities were found in 37 cases. The numeral abnormalities were more common than structural abnormalities, and most of the numeral abnormalities were aneupoidies. In turn, they were trisomy 16, 45,X, trisomy 22, trisomy 2, trisomy 21, trisomy 15. The most common structural abnormality was balanced translocation, including Robersonian translocation. Female embryoes accounted for 61.02%(1080/1770) miscarriages and for 57.4%(596/1770) of chromosomal abnormalities, while male embroyes acoounted for 61.02%(1080/1770),57.4%(596/1770)respectively. The proportion of female embryoes was higher than male embryoes. The median age of the patients was 30 years old(16-46 years old). As the maternal age increased, the proportion chromosomal abnormalities increased. The incidence of chromosomal abnormalities in the advanced age group (≥35 years) was 68.38%(240/351), which was significantly higher than that in the younger group (56.24% ,798/1419; χ2=17.10, P<0.01). Conclusions Embryonic chromosomal abnormalities are the most common cause of early spontaneous miscarriage. The abnormalities centralize in some karyotypes. There is certain relationship between maternal age and the incidence of miscarriage, as well as the embryonic gender. Chorionic villus cell culture and karyotype analysis are helpful in finding the cause of miscarriage and counsel the patients.