OBJECTIVETo determine the compound Daqiqi decoction( CDQD) combined cisplatin plays on the cell apoptosis of subcutaneous transplanted ovarian cancer in nude mice, to provide theory evidence for clinical treatment.

METHODMaking the models of subcutaneous transplanted ovarian cancer in nude mice, and divide the 40 mice with tumor into 5 groups (n = 8), the model control group, CDQD low dosed group, CDQD high dosed group, cisplatin group, cisplatin combined CDQD group, killing all the mice after 3 weeks' treatment and stripping tumors. Measure the volume and weight of the tumor and calculate tumor growth curve damps, the inhibition rate. Examining the expression of the Bcl-2(B-cell leukemia /lymphoma 2) and Bax (Bcl-2 associated x protein) mRNA and protein by the RT-PCR and the Western blot.

RESULT(O)The tumor weight shows that there was certain lighter effect in each CDQD group, and compared with cisplatin groups has no statistical significance, the cisplatin combined CDQD group is obviously lighter than that of the other group(P <0.01). The tumor growth curve damps shows that compared with model control group, the treatment group tumors had some extent narrowing (P < 0.01). (2) RT-PCR results: Bax (Bcl-2 associated x protein) mRNA expression shows that compared with model control group, the treatment group has increased (P < 0.01), and the cisplatin combined CDQD group compared with the other group is the highest, there was significant difference with the rest of the treatment group (P < 0.01). In the Bcl-2 mRNA express lowest in cisplatin combined CDQD group (P < 0.01), there has no difference between CDQD high dose group and cisplatin group. (Western blot shows: compared with model control group, the Bax protein of treatment group has increased expression (P < 0.01), the expression of Bax protein in cisplatin combined CDQD group is the highest(P < 0.01). There has no difference between CDQD high dose group and cisplatin group. The Bcl-2 protein expression of the cisplatin combined CDQD group is the lowest (P < 0.01), there has no difference between CDQD high dose group and cisplatin group.

CONCLUSIONThe effect of CDQD on subcutaneous transplantation ovarian tumor has promoting apoptosis function, its mechanism may be related to downgrade the Bcl-2 expression, higher expression of bax, stimulation on the apoptosis of tumor cells; cisplatin combined CDQD have synergistic effect.

Animals ; Cell Line, Tumor ; Cisplatin ; administration & dosage ; Disease Models, Animal ; Drug Therapy, Combination ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Ovarian Neoplasms ; drug therapy ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

Objective To explore the effects of Compound Malt Pill on gene expressions of androgen receptor (AR) and insulin receptor (INSR) in polycystic ovarian syndrome (PCOS) rats; To discuss its relevant mechanism of action.Methods Letrozole was used to induce and establish rat models. Rats were randomly divided into normal group, model group, Diane-35 group, low-, medium- and high-dose Compound Malt Pill groups. The normal group and model group received normal saline by gavage; the Diane-35 group was given Diane-35 by gavage; Compound Malt Pill groups were respectively given different concentrations by gavage for 21 d. The levels of serum testosterone (T) and insulin were measured by ELISA, and the gene expressions of AR and INSR were measured by RT-PCR. Results Compared with normal group, the serum T, insulin and the gene expressions of AR and INSR in model group increased significantly (P<0.05,P<0.01); compared with model group, the serum T, insulin and the gene expressions of AR and INSR in low-, medium- and high-dose Compound Malt Pill groups decreased significantly (P<0.05, P<0.01).Conclusion Compound Malt Pill can adjust endocrine-metabolic disorder in PCOS rats.

OBJECTIVETo evaluate the effect mechanism of warm acupuncture combined with auricular point sticking therapy and its efficacy on insomnia by monitoring the level of brain neurotransmitters in the insomnia patients.

METHODSOne hundred and thirty patients with insomnia were randomized into an observation group and a control group, 65 cases in each one. In the observation group, based on the treating principle of warming yang and benefiting qi, acupuncture was applied to Xinshu (BL 15), Pishu (BL 20), Shenshu (BL 23), Yaoyangguan (GV 3), Baihui (GV 20), Neiguan (PC 6) and Shenmen (HT 7). Warm acupuncture was supplemented at Xinshu (BL 15), Pishu (BL 20), Shenshu (BL 23) and Yaoyangguan (GV 3). The treatment was given once every day. In the control group, estazolam tablets, 0. 5 to 1 mg were prescribed for oral administration, 30 min before sleep at night. The treatment of 14 days was taken as one session and 2 sessions were required in the two groups. The encephal of luctuograph technology was used to observe the sleep quality and brain neurotransmitters before and after treatment in the two groups and the efficacy was compared between the two groups.

RESULTSThe total effective rate was 83.1% (54/65) in the observation group and 87.7% (57/65) in the control group. The efficacy was similar between the two groups (P > 0.05). In the observation group, after treatment, 5-HT and GABA/Glu were all increased compared with those before treatment (P < 0.05, P < 0.01) and norepinephrine (NE) was reduced compared with that before treatment (P < 0.05). The level of each index did not change significantly before and after treatment in the control group (all P > 0.05). The regulations of 5-HT, GABA/Glu in the observation group were superior to those in the control group (all P < 0.05).

CONCLUSIONThe combined therapy of warm acupuncture and auricular point sticking method for warming yang and benefiting qi effectively improves brain neurotransmitters and essentially improves sleep quality of the patients with insomnia differentiated as yang deficiency pattern.

Acupuncture Points ; Acupuncture Therapy ; Acupuncture, Ear ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neurotransmitter Agents ; metabolism ; Serotonin ; metabolism ; Sleep Initiation and Maintenance Disorders ; metabolism ; therapy ; Treatment Outcome ; Yang Deficiency ; metabolism ; therapy ; Young Adult ; gamma-Aminobutyric Acid ; metabolism

Objective To study the role of JAK-STAT singal transduction pathway in the interstitial fibrosis of unilateral ureter obstruction (UUO)mice. Methods Mice UUO model was established and the phosphorylation of JAK-STAT was examined at day 1,4,7 and 14 after ligation of the ureter.Mice in the treatment group were treated with daily injection of selective JAK2 inhibitor AG490 starting 2 h before ureter ligation until sacrifice while vehicle alone was given to mice in the model control group.Mice were sacrificed at day 14 after the establishment of model.Renal tubular lesion and interstitial fibrosis were assessed on paraffin section.Immunohistochemistry was used to detect renal macrophage infihration and α-SMA expression.The expression of collagen Ⅲ and MCP-1 mRNA was measured by RT-PCR.Phosphorylation of JAK2and STAT1 was examined by Western blotting. Results JAK2-STAT1 signaling transduction pathway was activated in UUO model.The activation of JAK2-STAT1 was closely correlated with the progression of renal injury,tubular histological lesions and interstitial fibrosis.AG490 treatment significantly inhibited the phosphorylation of JAK2 and STAT1 (P＜0.01).AG490 treatment also significantly reduced tubular lesions[(21.7 ±1.7)% vs (49.4±1.0)%]and interstitial fibrosis(1.0±0.1 vs 2.3±0.2),α-SMA expression(0.9±0.1 vs 2.1±0.2)and maerophage accumulation[(13.3±1.6)cells/HPF vs (34.4±1.0)cells/HPF](all P＜0.01).In addition,AG490 significantly inhibited the expression of collagen Ⅲ and MCP-1 mRNA. Conclusion JAK-STAT signaling plays an important role in renal tubulointerstitial inflammation and fibrosis.

Objective To investigate the role of heme oxygenase-1(HO-1)in the protective actions of curcumin against ethanol-induced oxidative damage.Methods Microsomal HO-1 enzyme activities were determined in rat primary hepatocytes pretreated by curcumin.AST,LDH release and hepatic oxidative/antioxidant status were measured with or without ZnPPⅨ/Hemin as a classic HO-1 inhibitor/inducer,respectively.Results Curcumin could induce the HO-1 expression and enzyme activity,which was correlated with antioxidant levels in hepatocytes.HO-1 induction reached a peak under administration of 15 μmol/L curcumin for 1 h.Conclusion HO-1 induction by curcumin is contributed to the heptoprotective effects against ethanol-induced oxidative damage.

Cooperation hydrogen production was carried out using Rhodopseudomonas sp. DT and Enterobacter aerogenes. The effects of the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes, culture temperature, and carbon source on the cooperation hydrogen production were investigated. The results suggested that cooperation hydrogen production rate was highly affected by the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes. The mixed bacteria of Rhodopseudomonas sp. DT and E. aerogenes with 1:1 initial ratio benefited to the cooperation hydrogen production, which led the hydrogen production rate and duration of gas production to 3.1 mol H2/mol glucose and 81 h, respectively. The pH dynamics analysis of culture medium further discovered that the pH of the mixed bacteria with 1:1 initial ratio changed from 6 to 7 smaller than other conditions, which was probably fitted to produce hydrogen. Furthermore, the mixed bacteria with 1:1 initial ratio had the higher hydrogen production efficiency at temperatures of 28?C and 37?C than at 20?C, and without any hydrogen production at temperature of 50?C. The carbon sources of glucose, succinate acid, malic acid could be used to produce hydrogen by the mixed bacteria. Even the soluble starch, unused by Rhodopseudomonas sp. DT, was also decomposed by the mixed bacteria to produce hydrogen with the conversion efficiency of 8.22%. The glucose was the optimal carbon resource, and the conversion efficiency could reach to 36.11%. The results, further, implied that the cooperation hydrogen production could enlarge the use of the carbon sources.

OBJECTIVETo investigate polymorphisms of killer cell immunoglobulin-like receptor gene (KIR) in renal transplant recipients from southern Zhejiang.

METHODSKIR genotypes were analyzed by PCR-SSP in 416 renal transplant recipients, and the genotype frequencies were compared with populations from Eastern China and worldwide.

RESULTSAll 16 known KIR genes were detected in the renal transplant recipients, and KIR2DL4, 3DL2-3, 3PD1 were found in all. As a pseudogene, 2DP1 has a high genotype frequency (99%). The frequencies of KIR2DL1, 2DL3, 3DL1, 2DS4 have ranged from 92.1% to 98.8%. Compared with 11 groups in Eastern China and other countries, the KIR2DL2 phenotype frequency was higher (34.6%) than those of Shanghai, Zhejiang and Jiangsu populations (P<0.05). Among 41 genotypes, three have not been reported previously. The most common genotype was AA1, with a frequency of 43.51%, which was significantly lower than those of Jiangsu and Northern Zhejiang.

CONCLUSIONRenal transplant recipients from southern Zhejiang share similar features with Eastern China Han population with regard to KIR polymorphisms, but also have unique frequencies for KIR genotypes.

Adolescent ; Adult ; Aged ; China ; Female ; Gene Frequency ; Genotype ; Humans ; Kidney Transplantation ; methods ; Male ; Middle Aged ; Polymorphism, Genetic ; Receptors, KIR ; genetics ; Young Adult

To investigate a baculovirus insect cell system for expressing an interferon alpha 2b (IFNa2b)/immunoglobulin G-4 (IgG4) Fc fusion protein, which has long-acting antiviral effects. Human IFNa2b and IgG4 Fc cDNAs were generated by molecular cloning and inserted into a baculovirus shuttle vector, which was then transposed into the DH10 Bac strain to form recombinant Bacmid-IFN/Fc. The Bacmid-IFN/Fc was transfected into High five insect cells, and expression of the IFN/Fc fusion protein was detected by Western blotting and its biological activity was assessed by the cytopathic effect inhibition method. The IFNa2b and IgG4 Fc cDNA fragments were successfully amplified by RT-PCR using human peripheral lymphocytes. After cloning into the baculovirus shuttle vector, pFastBac1, and transforming into DH10 Bac competent cells, screening identified positive clones carrying the recombinant Bacmid-IFN/Fc. A Bacmid-IFN/Fc clone was successfully transfected into the High five insect cells and packaged into the baculovirus for expression of the IFN/Fc fusion protein. Western blotting revealed that the fusion protein expression was specific, and yielded a protein of 45 kD in size. The in vitro antiviral activity of the IFN/Fc fusion protein was 580 IU/mL. A novel IFN/Fc fusion protein was successfully generated using a baculovirus insect cell system, which may prove useful for providing future experimental data for development of a new long-acting interferon to treat chronic viral hepatitis.
Animals ; Antiviral Agents ; metabolism ; Baculoviridae ; genetics ; Cell Line ; Cloning, Molecular ; Gene Expression ; Gene Fusion ; Genetic Vectors ; Humans ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; Immunoglobulin G ; biosynthesis ; genetics ; Insecta ; Interferon-alpha ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

BMP6 is a member of TGF-beta superfamily, represent more effective osteogenic activity. Two recombinant plasmids were constructed to expression rhBMP6 in mammalian cells, one contained the cDNA encoding the signal peptide, propeptide and mature peptide of human BMP6, wich was named pcDNA-BMP6, the other contained the recombinant DNA encoding the signal peptide, propeptide of human BMP2 and the mature peptide of BMP6, which was named pcDNA-BMP2/6. Transient expression in Cos7 cells demonstrated that the pcDNA-BMP2/6 produced more rhBMP6 than pcDNA-BMP6. For stable expression, the CHO-dhfr- cells were transfected with pcDNA-BMP2/6 and pSV2-dhfr, then screened by G418 and treated with MTX for targeting gene amplification. The partially purified rhBMP6 by heparin affinity chromatography was shown to possess bone induction activity tested by the induction of alkaline phosphatase activity in C2C12 cells.
Alkaline Phosphatase ; metabolism ; Animals ; Blotting, Western ; Bone Morphogenetic Protein 2 ; genetics ; Bone Morphogenetic Protein 6 ; genetics ; metabolism ; pharmacology ; CHO Cells ; COS Cells ; Cell Line ; Cercopithecus aethiops ; Cricetinae ; Cricetulus ; Gene Expression ; Humans ; Myoblasts ; cytology ; drug effects ; enzymology ; Protein Precursors ; genetics ; Protein Sorting Signals ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Purpose To explore the mutation of CD79B and MyD88 in primary testicular diffuse large B cell lymphoma and their significance.Methods Histopathologic features were observed in 15 cases of primary testicular diffuse large B cell lymphoma and immunophenotype was analyzed by immunohistochemical staining (IHC).Sanger sequencing was used to detect CD79B Y196 and MyD88 L265 mutation in these cases.The relationship between CD79B,MyD88 mutation and the clinicopathological features was analyzed.Results Immunophenotypically,15 cases were non germinal center B cell type.CD79B (Y196) mutation was detected in 4 cases (26.7％).For MyD88,L265 mutation was found in 7 cases (46.7％).CD79B and MyD88 mutations were found in 3 cases.The followup information was obtained in 8 patients.No association was found between CD79B,MyD88 mutation and outcome of patients.Conclusion Primary testicular diffuse large B cell lymphoma of non germinal center B cell type is a rare aggressive B cell lymphoma with poor prognosis and poor response to chemotherapy.CD79B,MyD88 gene mnutation was detected in Chinese patients with frequency of 26.7％ and 46.7％ respectively.It is possible for molecular targeted therapy of the primary testicular diffuse large B cell lymphoma on the basis of high frequency of CD79B and MyD88 gene mutation.