Aim To evaluate the antinociception of ω-SO3,a novel Omega-superfamily conotoxin,in rat formalin test.Methods Potency and duration of ω-SO3 antinociception compared with morphine were investigated in rat formalin test after acute intrathecal administration.Development of tolerance or cross tolerance to analgesia of ω-SO3 and morphine was tested in formalin test after chronic intrathecal administration.Locomotor activity of rat after acute intrathecal administration was tested to evaluate possible central side effects.Results In rat formalin test after intrathecal bolus injection,ω-SO3 produced dose-and time-dependent antinociception by suppressing acute(ED_(50),1.79 ng·g~(-1))and tonic phases(ED_(50),0.41 ng·g~(-1)),which was approximately 10-fold potency and twice longer-acting of morphine in blocking tonic phase responses.After repeated intrathecal injections twice daily for 5 consecutive days,ω-SO3 produced analgesia without loss of potency whereas morphine produced analgesia tolerance in rat formalin test;further,ω-SO3 still produced potent analgesia in morphine-tolerant rats.No changes in motor function were seen in rats receiving above antinociceptive doses.Conclusion sNovel ω-SO3 produces potent and long-acting spinal antinociception without observable motor dysfunction and after chronic intrathecal administration.ω-SO3 produces neither tolerance nor cross-tolerance to morphine analgesia.

M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

Objective:To investigate whether the omental-adipose stromal cells (O-ASCs) exposing to gastric cancer-conditioned medi-um (CM) could be inducted to differentiate into carcinoma-associated fibroblasts (CAFs) and the effect of ERK signaling pathway in the process. Methods: We identified O-ASCs by examining their ability to differentiate osteogenic and adipogenic lineages and through flow cytometry. O-ASCs were co-cultured with MGC803 and SGC7901CM. The expression of CAFs markers (α-SMA, FSP-1, and vimentin) and paracrine factors (VEGFA, TGF-β1, FAP, and SDF-1) were evaluated by RT-PCR and Western blot. In vitro cultures of O-ASCs were divided into three groups:the control, SGC7901-CM, and SGC7901-CM+U0126 groups. Cells were collected after 12 h. West-ern blot was performed to evaluate the expression ofα-SMA, FSP-1, ERK, and p-ERK1/2. Results:The primary cells were O-ASCs. The expression levels of CAFs markers (α-SMA, FSP-1, and vimentin) and O-ASC paracrine factors (VEGFA, TGF-β1, FAP, and SDF-1) clearly in-creased (P<0.05). In comparison with the control, the expression of ERK in SGC7901-CM group did not change (P>0.05), while the ex-pression of p-ERK1/2,α-SMA, and FSP-1 significantly improved (P<0.05). Comparison of SGC 7901-CM+U0126 and SGC 7901-CM groups showed that the expression levels of ERK had no statistical difference (P>0.05), while the expression levels of p-ERK1/2,α-SMA, and FSP-1 decreased (P<0.05). Conclusion:O-ASCs participate in the peritoneal metastasis of gastric cancer through differentiation by CAF and paracrine factors. The ERK signaling pathway is important in the differentiation of O-ASCs towards CAFs.

Objective To investigate the radiosensitizing effects of dihydroartemisinin(DHA)on human HeLa cells of cervical cancer irradiated by X rays.Methods Cell growth kinetics was determined using MTF assay.Cell survival was analyzed by elonogenic assay.The change of cell cycle and apeptosis was measured by flow cytometry.Results Dihydroartemisinin inhibited the growth of HeLa cells of human cervical cancer and showed a dose-dependent and time-dependent manner.Dihydroartemisinin(20 μmol/L)showed the radiosensitizing effects on HeLa cells,and the sensitizing enhancement ratio(SER)was 1.47.Dihydroartemisinin abrogated radiation-induced G2 arrest of the tested HeLa cells,the G2 ratio of medicine + radiation group dechned from 73.58% to 48.31%.Dihydroartemisinin enhanced the apoptosis of HeLa cells by X-irradiation,the apoptosis rates of medicine + radiation group significantly increased from 29.46%,48.04%,70.21% to 45.79%,66.36% and 79.58%,respectively for 2,4 and 6 Gy.Conclusions Dihydroartemisinin could increase the radiesensitivity of HeLa cells of human cervical cancer.Abrogation of radiation-induced C2 arrest could be part of the mechanism.

ObjectiveTo assess the effects of rehmannia and storesin on minimal hepatic encephalopathy(MHE) in rat model.MethodsMHE rat model was induced by carbon tetrachloride (CCl4) intragastric administration. The effects of rehmannia and storesin on spontaneous movement and learning and memory function of model animals were evaluated with open field test and Morris water maze in 3 dose groups. Lactulose was used in the positive group.ResultsThe spontaneous movement and the spatial learning and memory ability of the model rats both improved significantly in the high dose group. Meanwhile, the serum level of alanine transarninase(ALT) and ammonia(Amm) also decreased in the high dose group.ConclusionRehmannia and storesin has therapeutical effect on MHE rat model.

Objective: The current study was designed to distinguish Phase G0/G1， S， G2/M cell groups in the whole cell cycle by DNA flow cytometry. Method: The authors developed a multiparameter flow cytometry of cyclin E＋ A/DNA for MOLT-4. Result: The data showed that the method could distinguish Phase G0, early G1, late G1, S, G2,and M cells. The method could be used in molecular cell biology, especially in cell kinetic study.

The clinical value of transvaginal ultrasound in clinical surgical treatment of cesarean scar pregnancy (CSP) was evaluated.The clinical data of 104 patients with CSP admitted at Tongji Hospital from 2013 to 2016 were collected and analyzed retrospectively,including the patients' age,gestational age,the size of gestational sac or uneven mass,the uterine scar thickness,β human chorionic gonadotropin (hCG) levels and so on.Of these 104 cases,30 cases were subjected to laparotomy,29 cases to laparoscopy,27 cases to hysteroscopy,16 cases to ultrasound-guided uterine curettage,and 2 cases to conservative treatment.The transvaginal ultrasound showed that uterine scar thickness and gestational sac or uneven mass size had significant difference (P＜0.05) among different surgical methods by comparatively analyzing the patients' data.It was suggested that transvaginal ultrasound may provide the valuable reference for choosing clinical surgical procedures for CSP.

OBJECTIVETo investigate the effects of hydroquinone (HQ) on expression of Polymerase eta (Pol eta) and DNA damage in human hepatic cells (L-02), and to explore the role and possible mechanism of Pol eta involved in the process of DNA damage-tolerance.

METHODSAfter L-02 hepatic cells were exposed to HQ with various concentrations (0, 5, 10, 20, 40, 80 and 160 micromol/L) for 24 h, cell survival rate was detected by MTT assay; DNA impairment was detected by single cell gel electrophoresis (SCGE); Real-time fluorescent quantitative PCR and Western blotting methods were used to measure the expression of Pol eta at the mRNA and protein level in L-02 hepatic cells exposed to HQ with various concentrations (0, 5, 10, 20, 40, 80 and 160 micromol/L).

RESULTSMTT assay showed that HQ with concentrations from 0 to 80 micromol/L had little effect on the survival rate of L-02 (P>0.05); whereas the survival rate of the group of 160 micromol/Lwas significantly higher than that of the control (P<0.01) after being treated with HQ for 24 h; the higher dose of HQ presented, the more degrees of DNA damage were produced. It was found that HQ in a low concentration (1-80 micromol/L) could induce the expression of Pol eta which was in proportion to the increasements of HQ concentration; the expression levels of mRNA and protein were reached to the maximum when treated with 80 micromol/L; the expression of Pol eta decreased (the relative quantity values were 2.32 +/- 0.16 and 1.20 respectively) once the concentration of HQ exceeded 160 micromol/L as compared with the group of 80 micromol/L, but it was higher than that of the control.

CONCLUSIONThis study suggested that Pol eta might involve in the process of DNA damage-tolerance induced by HQ in the hepatic cells.

Cell Survival ; drug effects ; Cells, Cultured ; DNA Damage ; drug effects ; DNA Repair ; DNA-Directed DNA Polymerase ; metabolism ; Hepatocytes ; drug effects ; metabolism ; Humans ; Hydroquinones ; adverse effects ; Mutagens

In the present study, the modulatory effect of AMPA receptors on gamma-aminobutyric acid (GABA) transporter current was investigated on enzymatically isolated horizontal cells of carp retina. The GABA transporter current elicited by 1 mmol/L GABA was decreased immediately after pre-application of AMPA (30 mumol/L or 3 mmol/L) for 50 s. Application of 10 mmol/L BAPTA in intracellular solution inhibited the suppression effect of AMPA on GABA transporter current. The suppression effect induced by co-application of 3 mmol/L AMPA and 3 mmol/L NMDA was similar to that of 3 mmol/L AMPA or 3 mmol/L NMDA alone. These results suggest that the activation of AMPA receptors inhibits GABA transporter-mediated current by affecting intracellular Ca(2+) processes in the retinal horizontal cells, which is identical with the modulatory effect of NMDA receptors on GABA transporters.
Animals ; Carps ; Egtazic Acid ; analogs & derivatives ; pharmacology ; GABA Plasma Membrane Transport Proteins ; metabolism ; Receptors, Ionotropic Glutamate ; metabolism ; Retinal Horizontal Cells ; metabolism ; gamma-Aminobutyric Acid ; pharmacology

OBJECTIVETo explore the effect of berberine on lipid metabolism disorder and lipid deposition in liver cells of non-alcoholic fatty liver disease (NAFLD) rats induced by high fat diet.

METHODSAfter one week adaptable feeding, 45 SPF level male SD rats were randomly divided into 3 groups, the normal control group, the model group, and the berberine group, 15 in each group. Except those in the normal control group, all rats were fed with high fat diet to prepare NAFLD model. As for rats in the berberine group, Berberine Hydrochloride was administered by gastrogavage. HE staining and oil red O staining were performed to identify the model after 8 weeks. Hepatocytes were isolated, and their activities and purities were tested by Typan blue staining and flow cytometry (FCM). Serum levels of TC, TG, HDL-C, and LDL-C were detected using automatic biochemical analyzer. mRNA expression levels of LXRα and FAS in liver cells were analyzed by Real-time quantitative polymerase chain reaction (PCR). Protein levels of LXRα and FAS in liver cells were examined by Western blot.

RESULTSThe NAFLD rat model was successfully established by high fat diet. The yields of purified liver cells in each rat were (6.0-7.5) x 10(8). The viability of isolated liver cells with purity over 90% (tested by FCM analysis) was higher than 95%. Compared with the normal control group,the expression of LXRα and FAS at mRNA and protein levels was higher in the model group (P < 0.01). Compared with the model group, the expression of LXRα and FAS at mRNA and protein levels was obviously down-regulated in the berberine group (P < 0.01).

CONCLUSIONSLXRα/FAS signaling pathway was one of important signaling pathways of NAFLD lipid metabolism disorders. Berberine could recover hepatocyte fatty deposits in NAFLD rats by adjusting the LXR/FAS signaling pathway of hepatocytes, which might be one of important mechanisms for fighting against NAFLD.

Animals ; Berberine ; therapeutic use ; Diet, High-Fat ; Down-Regulation ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Liver ; Hepatocytes ; Lipids ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction