OBJECTIVETo explore the effect of Bushen Tiaojing Recipe (BTR) on the quality of oocytes, reproductive hormones, and the expression of bone morphogenetic protein-15 (BMP15) of in vitro fertilization-embryo transfer (IVF-ET) patients.

METHODSSixty infertility patients who prepared for IVF-ET were assigned to two groups according to the treatment order, the treatment group [20 cases, treated with BTR + controlled ovarian hyperstimulation (COH)] and the control group (treated with COH alone, 40 cases). Age, the time limit for infertility, basal follicle-stimulating hormone (bFSH) concentration, usage days and the dosage of gonadotropins (Gn), serum levels of estradiol (E2), luteotropic hormone (LH), and progesterone (P) on the HCG injection day, the number of retrieved occytes, the fertilization rate, the number of embryos, the high quality embryo rate, and the clinical pregnancy rate were compared. Concentrations of follicle-stimulating hormone (FSH), LH, E2, testosterone (T), and P in the follicular fluid were detected via chemiluminescence microparticle immunoassay. The mRNA and protein expression of BMP-15 in mature granulosa cells was detected by real-time fluorescent PCR and Western blot.

RESULTSThirty-two patients were pregnant and the total pregnancy rate was 53.3%. Of them, 19 were pregnant and the total pregnancy rate was 47.5% in the control group, while 20 were pregnant and the total pregnancy rate was 65.0% in the treatment group. But there was no statistical difference between the two groups (P > 0.05). Compared with the control group, the Gn dosage was lower and the high quality embryo rate was higher in the treatment group, showing statistical difference (P < 0.05). There was no statistical difference in serum concentrations of E2, LH, or P on the HCG injection day, the number of retrieved oocytes, or the fertilization rate (P > 0.05). Compared with the control group, FSH concentrations in the follicular fluid were significantly lower and LH concentrations were significantly higher in the treatment group (P < 0.05). The LH concentrations in the follicular fluid were significantly higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05).There was no statistical difference in E2, T, or P concentrations (P > 0.05). The mRNA and protein expression of BMP-15 in granulosa cells was higher in the treatment group than in the control group (P < 0.05). It was also higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05).

CONCLUSIONDuring the IVF-ET process, BTR could elevate the quality of oocytes, and increase the sensitivity of ovarian follicles to exogenous Gn, which was correlated with the mRNA and protein expression of BMP-15 in granulosa cells, and changing concentrations of FSH and LH.

Adult ; Bone Morphogenetic Protein 15 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Embryo Transfer ; Estradiol ; blood ; metabolism ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; metabolism ; Follicular Fluid ; metabolism ; Humans ; Luteinizing Hormone ; blood ; metabolism ; Oocytes ; drug effects ; Pregnancy ; Progesterone ; blood ; metabolism ; Testosterone ; metabolism ; Young Adult

Objective To explore the association of Gly71Arg mutation in gene of bilirubin uridine 5'-diphosphate-glucuronosyltransferase(UGT1A1)and neonatal jaundice in Beijing city Han population.Methods The genotypes and alleles of the Gly71 Arg polymorphism for UGT1A1 gene were identified by polymerase chain reaction-restricted fragment length polymorphism assay in infants of Beijing city Han population of China,including 96 infants with neonatal jaundice[serum bilirubin(307.06?38.5)?mol/L,indirect bilirubin(292.9?35.9)?mol/L] and 101 healthy control infants [serum bilirubin(131.2?42.1)?mol/L,indirect bilirubin(126.3?39.7)?mol/L].The genotypes and allele frequencies of the polymorphism were compared between infants with neonatal jaundice group and healthy infant group(control group).The effect of polymorphism in infants with neonatal jaundice group on serum bilirubin level were analyzed.Results There were significant differences in genotypes distribution in Gly71Arg polymorphism for UGT1A1 gene between the 2 groups(?2=9.47 P=0.002).Compared with control group,neonatal jaundice group had significantly higher Arg allele frequency in the polymorphism for UGT1A1 gene(?2=10.34 P=0.001).There were independent effects of Gly71Arg mutation in the gene on serum bilirubin level in neonatal jaundice group,at the carriers of homozygote of the Arg allele of Gly71Arg polymorphism had higher serum bilirubin levels compared to carriers of heterozygote of the Arg allele of the polymorphism and non-carriers of the Arg allele of the polymorphism(Pa

A clinically reliable non-invasive test for endometriosis is expected to reduce the diagnostic delay. Although varieties of biomarkers have been investigated for decades, and cancer antigen-125, cancer antigen-199, interleukin-6, and urocortin were the most studied ones among hundreds of biomarkers, no clinically reliable biomarkers have been confirmed so far. Some emerging technologies including "omics" technologies, molecular imaging techniques, and microRNAs are promising in solving these challenges, but their utility to detect endometriosis has yet to be verified. New combinations of researched indicators or other non-invasive methods and further exploration of the emerging technologies may be new targets and future research hotspots for non-invasive diagnosis of endometriosis. In conclusion, researches of biomarkers for the detection of endometriosis are still ongoing and may benefit from novel molecular biology, bioinformatics methods and a combination of more diverse monitoring methods. Though it will be a daunting task, the identification of a specific set of diagnostic biomarkers will undoubtedly improve the status of endometriosis.

To study still further the activity of CNTF mutant designed by computer molecular modling,the methods of dissociated cultures of chick dorsal root ganglion、TF-1 prolification and the normal mice'weight loss tests weve used.The results indicated that the mutant protein promoted the survival of dorsal root ganglion、induced TF-1 prolification and made the normal mice lose weight,decrease appetite and reduce fat index.The weight loss effect was dependant with its administration dosage,ED50 was 150.986?g/kg/d.To TF-1,the specific activity reached 2.0?106U/mg against international reference reagent.In a word,CNTF mutant had excel bioactivity.So it provided clues for its development and application.

Alternol is purified from fermentation productions of microorganisms named as Alternaria alternata var. monosporus. The research is to investigate the apoptosis-inducing effect of alternol on mouse lymphocyte leukemia (L1210) cells and the possible mechanisms. MTT method was used to evaluate the viability of L1210 cells. Apoptosis of L1210 cells was detected by morphological assessment, DNA electrophoresis assay and flow cytometry. Western blotting analysis was carried out to determine the apoptosis-related proteins. Proliferation inhibition of L1210 cells by alternol was found remarkably in a dose-dependent manner. When treated with alternol, apoptotic morphological features of L1210 cells were observed by fluorescent microscopy (AO/EB) and the apoptosis rate was also elevated in a time-dependent manner. After treatments with various concentrations of alternol for 48 h, DNA laddering appeared. The increase of reactive oxygen species (ROS) production was found after cells were exposed to alternol for 6 h, while the decrease of mitochondrial transmembrane potential (delta psi m) was not found until cells were exposed to alternol for 24 h. Furthermore, the level of Bcel-2 and Bcl-2/Bax was down-regulated, while the level of caspase-3 and caspase-9 but not caspase-8 was up-regulated when alternol was added for 72 h. In summary, the results suggested that alternol could inhibit the proliferation of L1210 cells and induce apoptosis of L1210 cells, which was mediated by mitochondria-dependent pathway.
Alternaria ; chemistry ; Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Heterocyclic Compounds, 4 or More Rings ; pharmacology ; Leukemia L1210 ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reactive Oxygen Species ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the effect of Bushen Tiaojing Recipe (BTR) on the expressions of drosophila mothers against decapentaplegic protein (Smadl), Smad5, Smad8, and Smad4 on human mural granulosa cells.

METHODSSixty-six patients undergoing in vitro fertilization-embryo transfer (IVF-ET) were randomly assigned to two groups in the ratio of 1:2, the treatment group and the control group. Twenty-three patients in the treatment group were treated with BTR and GnRHa/FSH/hCG, while forty-three patients in the control group were treated with GnRHa/FSH/hCG. The mRNA expressions of Smad1, Smad5, Smad8, and Smad4 on mural granulosa cells of the mature follicle were detected by real-time PCR on the ovum retrieval day. The expressions of Smad1, Smad5, Smad8, and Smad4 at the protein level were observed using cell immunofluorescence method.

RESULTSThe mRNA and protein expressions of Smadl in the granulosa cells were significantly higher in the treatment group than in the control group (P <0.05). There was no statistical difference in the mRNA and protein expressions of Smad5, Smad8, and Smad4 between the two groups.

CONCLUSIONThe mechanisms of BTR for improving the pregnancy rate and the ovarian functions might be correlated with up-regulating mRNA and protein expressions of Smadl of human mural granulosa cells.

Adult ; Drugs, Chinese Herbal ; pharmacology ; Female ; Granulosa Cells ; drug effects ; metabolism ; Humans ; Ovarian Follicle ; cytology ; Signal Transduction ; Smad1 Protein ; metabolism ; Smad4 Protein ; metabolism ; Smad5 Protein ; metabolism ; Smad8 Protein ; metabolism

OBJECTIVETo explore the antibacterial activity of amoxycillin sodium and clavulanate potassium (trade name: Anqi) in vitro and the pharmacoeconomics in the therapy of acute respiratory infection.

METHODSMinimal inhibition concentration (MIC), minimal bactericidal concentration (MBC) and bactericidal curve of amoxycillin sodium and clavulanate potassium against common pathogens were determined and compared with some other same kind of antibiotics without beta-Lactamase inhibitor. Eighty cases diagnosed as respiratory infection were randomly divided into 4 groups: group 1 was treated with i.v. Anqi; group 2 was treated with i.v. Anqi and oral consecutive strategy; group 3 was treated with iv ampicillin and sulbactam; group 4 was treated with i.v. cefuroxime. The clinical therapeutic effects were observed and cost-effectiveness analyzed.

RESULTSIn terms of MIC, MBC and bactericidal curve of 135 bacterial strains, Anqi was superior to the other same-kind antibiotics without beta-lactamase inhibitor, this effect was especially obvious on Klebsiella pneumoniae and Escherichia coli which can produce extended spectrum beta-lactamases (ESBLs). The cost-effectiveness of the consecutive therapy group was the best.

CONCLUSIONAnqi has a wide antimicrobial spectrum and strong effect on the bacteria producing ESBLs, the consecutive therapy strategy should be clinically recommended.

Amoxicillin ; pharmacology ; therapeutic use ; Anti-Bacterial Agents ; pharmacology ; therapeutic use ; Bacteria ; drug effects ; Child ; Child, Preschool ; Clavulanic Acid ; pharmacology ; therapeutic use ; Dose-Response Relationship, Drug ; Drug Therapy, Combination ; therapeutic use ; Female ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Respirovirus Infections ; drug therapy ; economics ; Treatment Outcome

Objective To carry on a survey on blood routine examination of coal-burning endemic fluorosis population in Bijie City,Guizhou Province in order to study their health status and problems.Methods Blood routine examination was performed in the residents in coal-fired pollution endemic fluorosis-endemic area, including the residents of the Changchun Village of Changcun Town(intervention group)whose stoves had been improved and of Shiba Village Yachi Town not improved in Bijie City,Guizhou Province.The indicators were including leukocyte(WBC),red blood cell(RBC),hemoglobin(Hb),hematocrit(HCT),tlle average hematocrit red blood cell volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration (MCHC),red blood cell distribution width-CV(RDW-CV),platelets(PLT).Results RBC,Hb,HCT,MCHC, PLT were(4.95±1.18)×1012/L,(138.46±15.90)g/L,(50.19±11.48)%,(284.90±48.73)g/L,(334.92± 119.34)×109/L for the male in the intervened group,and they were(4.02±0.47)x 1012/L,(131.00±15.90)g/L, (40.90±7.60)%,(323.14±41.95)g/L,(280.79±100.34)× 109/L in non-intervention group,respectively. Inter-group comparison,the difference was statistically significant (U = 7.72,3.50,7.12,6.28,3.66,P ＜ 0.01). RBC, HCT,MCV,MCH,MCHC,RDW-CV,PLT were respectively(4.75±1.20)×1012/L,(46.91±11.20)%,(99.30± 6.88)fl,(28.10±8.66)pg,(275.61±54.49)g/L,(16.95±1.63)%,(351.23±150.37)×109/L for the female in the intervened group,and were (3.85±0.65)×1012/L,(38.80±6.60)%,(100.80±7.00)fl,(33.10±5.40)pg, (327.14±44.52 ) g/L,(16.60±1.58) %,(279.40±98.07)×109/L in the group un-intervened. Inter-group comparison found that there was a significant difference(U = 8.92,10.72,2.04,6.61,9.82,2.06,5.39,P ＜ 0.001 or 0.05) and the abnormal rate of RBC and Hb in non-intervention group[ 32.62% (92/282),16.67%(47/282)] was higher than that in the intervention group[9.73%(29/298) ,6.71%(20/298),x2 = 45.992,14.054,P ＜ 0.01 ) ]. Conclusion Experiment group has better results of blood routine test compared to non-intervention group,especially of anemia.

ObjectiveTo investigate the effect of the Yap1 gene and tanshinone ⅡA on the proliferation, migration, and invasion abilities of Huh-7 hepatoma cells. MethodsA total of 10 pairs of human hepatocellular carcinoma (HCC) samples and adjacent tissue samples were collected in Zhongnan Hospital of Wuhan University from June 1 to December 1, 2019. Quantitative real-time PCR and Western blotting were used to measure the expression of the Yap1 gene and phenotype-related molecules. MTT cell proliferation detection reagent was used to measure the inhibition rate of cell proliferation after the treatment with different concentrations of tanshinone ⅡA. Western blotting was used to measure the changes in the expression of apoptosis-and migration-related markers after different interventions. Flow cytometry and Transwell assay were used to measure apoptosis and cell migration and invasion abilities. The data of 375 cases of liver cancer and 50 cases of relatively normal liver tissue samples were downloaded from The Cancer Genome Atlas, including clinicopathological information. The t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. ResultsIn 8 of the 10 pairs of HCC samples and adjacent tissue samples, HCC samples had significantly higher expression of Yap1 than the adjacent tissue samples. Compared with the normal human liver epithelial cells L02, the Huh-7 and HCCL-M3 hepatoma cells had a significant increase in the expression of Yap1. The silencing efficiency of si-Yap1-3 transfection reached 87.004% at the protein level. MTT results showed that tanshinone ⅡA effectively inhibited the proliferation of Huh-7 cells, with a half inhibitory concentration of 8.683 μmol/L. After the cells were treated with si-Yap1-3 and tanshinone ⅡA, there was an increase in the expression of the downstream marker for proliferation and migration E-cadherin and a reduction in the expression of vimentin, and the results of Transwell assay showed that compared with the si-NC group, the tanshinone ⅡA+si-Yap1-3 group had significant reductions in the migration and invasion abilities of Huh-7 cells (migration: 43.19±2.88 vs 132.20±10.03, t=8.527, P=0.001; invasion: 53.95±4.20 vs 179.10±11.11, t=4.484, P=0.011). The group treated with si-Yap1-3 and tanshinone ⅡA had an increase in the expression of the apoptosis-related marker Bax and a reduction in the expression of Bcl-2, as well as a significantly higher early apoptosis rate than the si-NC group (2598% vs 9.21%, χ2=4.078, P＜0.05). ConclusionOncogene Yap1 silencing combined with tanshinone ⅡA can promote the apoptosis of Huh-7 hepatoma cells and inhibit their migration and invasion, which can provide certain guiding significance for clinical medication.

OBJECTIVETo investigate the effect of cytochrome P450 isozymes on aristolochic acid induced cytotoxicity on renal proximal tubular epithelial cell (cell line HK-2).

METHODHuman renal tubular cells (cell line HK-2), were treated with aristolochic acid (AA) alone or in combination with cytochrome P450 isozymes inhibitors, including alpha-naphthoflavone (CYP450 1A1 and 1 A2 inhibitors), ketoconazole (CYP450 3A4 inhibitor), sodium diethyldithiocarbamate (CYP450 2A6 and 2E1 inhibitors), quinidine (CYP450 2D inhibitor), alpha-lipoic acid (NADPH: P450 reductase inhibitor), sulfaphenazole (CYP450 2C inhibitor) in the presence or absence of liver microsome(S9). The inhibition of cell proliferation rate was studied by MTT assay and the lactate dehydrogenase release rate was determined with continuous monitoring method.

RESULTAA inhibits cell proliferation and promotes the release of LDH over the range of 12.5-100 mg x L(-1), in a dose-dependent manner. Addition of S9 into the culture system reduced AA cytotoxicity, with the cell proliferation inhibition reducing and the release of LDH decreasing (AA + S9 group vs the same concentration of AA alone group, P < 0.05). In the absence of S9, ketoconazole or alpha-naphthoflavone has no obvious effect on AA cytotoxicity, however,under the conditions of adding S9, ketoconazole or alpha-naphthoflavone enhances AA cytotoxicity. Other inhibitors of CYP450 isozymes has no distinct effect on AA cytotoxicity.

CONCLUSIONThe microsomal enzyme of Liver can reduce the AA cytotoxicity, and CYP450 3A, CYP450 1A may be the major cytochrome P450 isozymes which impact AA cytotoxicity.

Animals ; Aristolochic Acids ; toxicity ; Cell Line ; Cell Proliferation ; drug effects ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Cytotoxicity, Immunologic ; drug effects ; Enzyme Inhibitors ; pharmacology ; Humans ; Kidney Tubules ; drug effects ; enzymology ; immunology ; Male ; Rats ; Rats, Wistar