AIM To explore the effects of sinomenine (Sin) on psychic dependence induced by morphine (Mor) in mice and its relation to the cAMP content of the cerebral cortex. METHODS A biased procedure induced the strong place preference. The effect of Sin (10, 30, 60 mg?kg -1 , ip) on the rewarding property of morphine was observed in a conditioned place preference(CPP) paradigm in mice,at the same time, the change of cAMP content in cerebral cortex was measured by the radioimmunoassay(RIA). RESULTS Sin significantly shortened the time of staying in morphine-pair compartment and lowered the cAMP level of mice. CONCLUSION Sin could attenuate effects of psychical dependence induced by Mor, its mechanism may be related to decrease of cAMP level in the brain, and Sin did not display psychical dependence in the experiment.

Diabetic rat model accompanied by insulin resistance was established by intraperitoneal injection of streptozocin.Following metformin treatment for 5 weeks,ELISA was used to detect the level of plasma secreted frizzled-related protein (SFRP) 5,and immunohistochemistry was used to detect the expression of p-JNK in the liver.insulin resistance(IR) and p-JNK were significantly increased in diabetic group compared with the control group (P＜0.05),while plasma SFRP5 level was reduced (P＜0.01).After metformin treatment,the plasma SFRP5 levels were significantly increased (P＜0.05),while IR and p-JNK was decreased (P＜O.05).Metformin may ameliorate insulin resistance via upregulating the SFRP5 expression of diabetic rats.

Objective To study of zoledronic acid in the treatment of multiple myeloma bone dis-ease clinical effect and detection of serum macrophage inflammatory protein (MIP)changes of primary mye-loma (mm)in patients with serum macrophage inflammatory protein levels and multiple myeloma bone dis-ease curative effect.Methods 48 cases of multiple myeloma bone disease patients were treated with VTD regimen chemotherapy were randomly and equally divided into two groups,one group (group A)chemother-apy intermission applied zoledronic acid 4 mg per month 1 time,treatment 2 course of treatment,observa-tion of curative effect and adverse reaction,another group (B group)declined to azole phosphonic acid treatment.Results Group of pain Solution of 16 cases were markedly effective,effective in 4 cases,4 ca-ses were ineffective,efficiency 83.3%.B group bone pain relieved markedly effective in 12 cases,effective in 4 cases,8 cases were ineffective,have efficiency 66.7%.A compared to the B,the curative effect was obvious (P <0.05).By enzyme linked immunosorbent assay for the detection of the patients with a,levels of peripheral serum MIP-1a and MIP-1 beta B two groups before and after treatment.Conclusions zole-dronic acid in the treatment of multiple myeloma bone disease effectively,can significantly improve the qual-ity of life in patients with MM patients serum MIP-1a and MIP-1 beta level and multiple myeloma tumor bone disease curative effect is negative correlation,used for evaluating the effect The reference index.

Background and purpose:Tumor suppressor gene P53 has long been studied in tumors, including breast cancer. More studies focused on the relationship between P53 and prognosis of breast cancer and found that P53 overexpression suggested a bad prognosis. However, the effect of P53 on early stage postmenopausal patients with ER-positive breast cancer has not been clariifed yet. This study was to investigate the role of P53 plays in aromatase inhibitor (AI) resistance among early stage postmenopausal patients with ER-positive breast cancer patients. Methods:A total number of 293 operable breast cancer patients who received surgical treatment during Jul. 2000 to Jul. 2006 in Fudan University Shanghai Cancer Center were enrolled into this study. All patients received AI treatment. The SPSS 12.0 software was used to estimate the survival rate. Univariate and multivariate analysis were also performed via above software. Results:The median follow-up time is 72 months (6-140 months). The 5 year disease free survival (DFS) of P53 positive and negative were 78%and 89%. The results showed that P53 overexpression (HR=1.729, 95%CI:1.038-2.880, P=0.035), pathological stage (HR=2.270, 95%CI:1.399-3.681, P=0.001);histological grade (HR=2.328, 95%CI:1.312-4.133, P=0.004); age (HR=1.988, 95%CI:1.511-2.617, P<0.005) were still the independent risk factors of recurrence and metastasis in breast cancer patients treated with AI. Conclusion:P53 overexpression correlated strongly with AI resistance in early stage postmenopausal patients with ER-positive breast cancer patients who were treated with AI and conifrmed the relevance of previously described prognostic factors. It is reasonable to take P53 expression into account when we evaluate the risk of breast cancer patients and decide the anti-cancer treatment strategy.

OBJECTIVETo observe the efficacy of Zaozhu Yinchen Recipe (ZZYCR) on non-alcoholic steatohepatitis (NASH) patients, and to explore its effect on serum free fatty acid (FFA) and tumor necrosis factor alpha (TNF-alpha).

METHODSTotally 120 patients with NASH were randomly assigned to the treatment group (60 cases, treated with ZZYCR, one dose per day) and the control group (60 cases, treated with Silibin Meglumine Tablets, 20 mg each time, thrice per day). The therapeutic course for all was 24 weeks. Serum levels of ALT and AST activities, TC and TG levels were detected before and after treatment. Peritoneal CT was performed in all patients, and CT ratios of liver and spleen calculated. NAFLD activity score (NAS) and degree of hepatic fibrosis were assessed using pathological examinations of liver tissue, and efficacy also evaluated. Serum contents of FFA and TNF-alpha were also detected.

RESULTSCompared with before treatment in the same group, activities of ALT and AST, serum levels of TC, TG, FFA, and TNF-alpha, NAS, scores of symptoms and signs all obviously decreased, degree of hepatic fibrosis was obviously improved in the two groups (P < 0.05, P < 0.01). These changes were more obviously seen in the treatment group (P < 0.05). After 24-week treatment, the total effective rate and total clinical efficacy were 80.00% (48/60 cases) and 85.00% (51/60 cases) in the treatment group, obviously higher than those in the control group [60.00% (36/60 cases) and 73.33% (44/60 cases) respectively], with significant difference (P < 0.05, P < 0.01).

CONCLUSIONZZYCR could improve the clinical efficacy of NASH patients, and its mechanism might be associated with inhibiting serum levels of FFA and TNF-alpha.

Objective: To evaluate the efficacy and side effects of priming regimen with pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in the treatment of initial treatment elderly patients with acute myeloid leukemia (AML). Methods: Thirty-five elderly patients with early-stage AML (non-M₃) who received pre-excitation chemotherapy in Yancheng Third People's Hospital from February 2015 to January 2019 were retrospectively analyzed. According to the different granulocyte colony-stimulating factor (G-CSF) in the chemotherapy regimen, 15 cases were in PEG-rhG-CSF group, 6 mg PEG-rhG-CSF was used alone on day 0 by subcutaneous injection; 20 cases were in recombinant human granulocyte colony-stimulating factor (rhG-CSF) group, 200 μg/m² rhG-CSF was used per day from day 0 to day 13 by subcutaneous injection, rhG-CSF was suspended or continued according to the number of white blood cells. In addition, both groups were given priming regimen with cytarabine and arubicin, or cytarabine and harringtonine. The efficacy and adverse reactions of the two groups were compared. Results: In the PEG-rhG-CSF group, there were 5 cases of complete remission, 6 cases of partial remission, 4 cases of non-remission, and 11 cases were effective. In the rhG-CSF group, there were 8 cases of complete remission, 7 cases of partial remission, 5 cases of non-remission, and 15 cases were effective. There was no significant difference in the efficacy between the two groups (χ ²= 0.012, P= 0.911). In terms of adverse reactions, the incidence of infectious fever, bone pain, duration of neutropenia, and duration of thrombocytopenia were not statistically significant (all P > 0.05). Conclusions In the pre-excitation chemotherapy for AML, the clinical efficacy and adverse effects of PEG-rhG-CSF are similar to rhG-CSF. However, the use of PEG-rhG-CSF can simplify the operation and reduce the pain and risk of local infection during chemotherapy.

Objective: To study the biological effects of the HPV16Z-Hsp65-E6/E7 fusion protein vaccine on the tumor associated with HPV16 infection. Methods: We tested the cellular immune responsive intensity to the vaccine by the lymphocyte proliferation and CTL response, and studied the therapeutic effect of the vaccine on mouse TC-1 cell transplanted cancer in vivo and the influence on mouse lifetime. Results: The spleen lymphocytes from the C57BL/6 mouse immunized by the Hsp65-E6/E7 vaccine could proliferate obviously in the presence of the protein and TC-1 tumor cell could be lysed specifically by immune activated lymphocytes in vitro. This animal therapeutic experiment in vivo showed that the vaccine suppressed the growth of TC-1 cell transplanted tumor remarkably. Conclusion: The recombined vaccine can induce specific cellular immune response in vitro and suppress HPV16 positive TC-1 tumor cell growth obviously in vivo.

OBJECTIVETo study the microstructue of acellular dermal matrix (ADM) and the attachment and proliferation of MG63 osteoblast-like cells on the membrane.

METHODSADM was experimental group and expanded polytetrafluoroethylene (e-PTFE) membrane was control group. Light microscope and scanning electronic microscope (SEM) were used to observe the microstructure of the two kinds of membrane. MG63 osteoblast-like cells were cultured on the two kinds of membrane and the blank group. The cell viability was checked by cell viability analyzer (CVA) and alkaline phosphatase (ALP) activity was detected by ELISA. SEM was used to observe the adhesion and proliferation of the MG63 osteoblast-like cells on the two kinds of membrane.

RESULTSThe ADM was composed of the tissular side and basal lamina side. The tissue side showed scale-shaped structure and the basal lamina side demonstrated many digitations and some pores of folliculus pili. The e-PTFE showed many tiny lined cracks in elliptic structure. The viability of the MG63 osteoblast-like cells on the two kinds of membranes and ALP measurement showed that there was no significant difference between each of the two membrane groups and the blank. The adhesion and proliferation of osteoblast-like cells on the two kinks of membranes were both well.

CONCLUSIONADM has no negative effects on the growth of osteoblast-like cells. ADM is biocompatibile and its microstructure is appropriate for guided bone regeneration materials.

In this study, Actin, 18S rRNA, PAL, GAPDH and CPR of Artemisia annua were selected as candidate reference genes, and their gene-specific primers for real-time PCR were designed, then geNorm, NormFinder, BestKeeper, Delta CT and RefFinder were used to evaluate their expression stability in the leaves of A. annua under treatment of different concentrations of Cd, with the purpose of finding a reliable reference gene to ensure the reliability of gene-expression analysis. The results showed that there were some significant differences among the candidate reference genes under different treatments and the order of expression stability of candidate reference gene was Actin > 18S rRNA > PAL > GAPDH > CPR. These results suggested that Actin, 18S rRNA and PAL could be used as ideal reference genes of gene expression analysis in A. annua and multiple internal control genes were adopted for results calibration. In addition, differences in expression stability of candidate reference genes in the leaves of A. annua under the same concentrations of Cd were observed, which suggested that the screening of candidate reference genes was needed even under the same treatment. To our best knowledge, this study for the first time provided the ideal reference genes under Cd treatment in the leaves of A. annua and offered reference for the gene expression analysis of A. annua under other conditions.
Artemisia annua ; drug effects ; genetics ; metabolism ; Cadmium ; pharmacology ; Plant Leaves ; drug effects ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; methods ; standards ; Reference Standards

Objective: To develop a therapeutic adjuvantfree protein vaccine against HPV16 which is closely related to cervical cancer of China. Method: First the E6/E7 gene by PCR technology from HPV16z virus strain was isolated in the highrisk cervical cancer area of Shanxi province of China in1990s, and again got the gene segment of Hsp65 from BCG by the same method, mutated the transforming codes in sequences of HPV 16 E6/E7 genes and thus constructed the expression vector pET28aHsp65E6/E7, expressed the Hsp65E6/E7 fusion protein in E.coli BL21(DE3) strain and researched optimal protein purification procedures. Results: The expression vector pET28aHsp65E6/E7 was constructed successfully and E6/E7 gene was mutated correctly. Hsp65E6/E7 fusion protein was renatured and purified on the affinity chromatography column simultaneously. The protein purity achieved 95% after the anionic exchange chromatograph purification. conclusions: This research laid a foundation for further functional study of the therapeutic adjuvantfree protein vaccine——Hsp65E6/E7.