In order to improve the expression level of segment of GABAA receptor a1 subunit in E. coli, growth conditions of the recombatant, which influence the final yield of protein expression, including growth medium, inoculation ratio, temperature, pH, rotation speed, inducing time and concentration of IPTG and so on, were studied in shaking flasks. The results indicated that, with 3% inoculation ratio, cultured 3.5 hours at 37℃, and then induced 5 hours by IPTG at 32℃, the yield of GABAA receptor protein was 95mg/L and the biomass was 3.25 g/ L. In contrast, using a 16 L stirred fermentor instead of shaking flasks, the highest level of the protein expression, 136mg/L with 4.95g/L of biomass, was achieved after fermenting 5.5 hours.

Adult ; Diverticulum ; complications ; Echocardiography ; methods ; Heart Diseases ; diagnosis ; Heart Ventricles ; pathology ; Humans ; Male ; Young Adult

Objective To analyze the examination results of external quality assessment (EQA),at all levels of iodine deficiency disorders (IDD) network laboratories in Ningxia Province and to further standardize and improve the laboratory,and to provide a reliable laboratory quality assurance for surveillance and control of IDD.Methods The examination results of EQA at all levels of IDD laboratories in Ningxia Province were statistically analyzed in accordance with the National Reference Laboratory (NRL) of IDD (2002-2011).Results Laboratory hardware equipment and technology at all levels met the testing requirements,and qualified rate of quality control increased year by year.Both of the response rate and qualification rate of urine iodine laboratories at provincial level were 100％ in the past decade.From 2005 on,the response rate of city laboratories had been 100％,and the qualification rate had been 100％ since 2007.The response rate and qualification rate of salt iodine laboratories at both the provincial level and the city level were 100％ in the past decade.The response rate of salt iodine laboratories at county level had been 100％ since 2004,and the qualification rate had been 100％ since 2009.Salt iodine and urinary iodine levels were fully qualified for the past three years at provincial,municipal and county levels.Conclusions All levels of IDD network laboratory in Ningxia Province runs good,EQA is fully qualified,and is able to provide a reliable laboratory quality assurance for surveillance and control of IDD.

A mixed bacteria culture F6 isolated from oil field wastewater can degrade petroleum hydrocarbons efficiently. The bacteria were suitable to treat oil-polluted wastewater of oil field. Simulated result treating oil-polluted wastewater in laboratory showed that after "XingyiLian" wastewater of Liaohe Oil Field was treated by fluidized-bed bioreactor system with the vehicle of activated carbon , the amount of oil and CODcr of the flow out water were decreased from 45mg/L to 4. 1mg/L and 470mg/L to 42mg/L separately , according with first class standards of Chinese Wastewater Discharge Regulation.

ObjectiveTo observe protective effects of agmatine (AGM) on inflammatory response and spleen immune function in mice with trauma.Methods Forty-eight adult male C57BL/6 mice were randomly divided into three groups (n= 16 each), including control group, model group (bilateral femoral fracture and removal of 35% of the total blood volume), and AGM group (trauma/hemorrhage & AGM 200 mg/kg). Eight mice in each group were sacrificed at 3 hours and 24 hours, respectively, after modeling, and blood samples and tissue homogenate of spleen and liver were collected. The contents of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-1β) in serum and liver tissue were determined with enzyme linked immunosorbent assay (ELISA). Serum aspartate transaminase (AST), alanine aminotransferase (ALT) and lactic dehydrogenase (LDH) were determined with automatic biochemistry analyzer. Spleen proliferation response stimulated with concanavalin A (ConA) was evaluated with methyl thiazolyl tetrazolium colourimetry (MTT).γ-interferon (IFN-γ) and IL-2 releases were determined with ELISA.Results Compared with control group, 3 hours after trauma/hemorrhage, the levels of serum TNF-α, IL-6, and IL-1β in model group were significantly elevated [TNF-α (ng/L): 145.38±31.50 vs. 23.06±11.14, IL-6 (ng/L): 496.94±50.76 vs. 47.13±17.47, IL-1β (ng/L): 321.31±43.02 vs. 29.25±16.24,allP< 0.01]. It was found that AGM treatment could alleviate the increase in serum pro-inflammatory mediators induced by trauma/hemorrhage, such as TNF-α (ng/L:111.56±25.47 vs. 145.38±31.50), IL-6 (ng/L: 412.56±44.33 vs. 496.94±50.76), IL-1β (ng/L: 273.38±45.25 vs. 321.31±43.02,P< 0.05 orP< 0.01). Twenty-four hours after trauma/hemorrhage, serum pro-inflammatory mediators were recovered to the levels in control group. There was no significant difference in TNF-α and IL-6 levels at 3 hours after trauma/hemorrhage among groups. Compared with control group, the expressions of liver TNF-α and IL-6 in model group were increased at 24 hours following trauma [TNF-α (ng/mg): 32.93±4.90 vs. 26.58±2.33, IL-6 (ng/mg): 11.20±1.66 vs. 8.38±0.89,bothP< 0.01]. However, AGM inhibited the level of TNF-α (ng/mg:28.92±3.16 vs. 32.93±4.90) and IL-6 (ng/mg: 9.03±1.28 vs. 11.20±1.66) in the liver as induced by trauma/hemorrhage (P< 0.05 andP< 0.01). At 24 hours after modeling, model group and AGM group had distinctly higher serum AST, ALT, LDH levels than those of control group [AST (U/L): 405.9±31.2, 245.7±22.1 vs. 128.2±15.9; ALT (U/L): 92.1±6.3, 51.6±5.0 vs. 30.1±3.2; LDH (U/L): 606.7±36.3, 478.7±25.3 vs. 384.0±16.6, allP< 0.01]. Nevertheless,the increase in serum AST, ALT and LDH was alleviated in AGM group (allP< 0.01). Meantime, trauma/hemorrhage produced a noticeable depression of proliferation of splenic cells and IFN-γ and IL-2 release stimulated with ConA compared with control group [proliferation rate: (40.97±4.13)% vs. (89.99±7.76)%, IFN-γ(ng/L): 91.6±12.3 vs. 353.2±21.5,IL-2 (ng/L): 53.4±6.4 vs. 91.0±12.2,allP< 0.01]. In contrast, AGM notably restored the capacity of proliferation response of splenic cells [proliferation rate: (74.86±5.75)% vs. (40.97±4.13)%, P< 0.01],enhanced the release of IFN-γ and IL-2 stimulated with ConA [IFN-γ (ng/L): 327.8±23.6 vs. 91.6±12.3, IL-2 (ng/L): 74.8±10.4 vs. 53.4±6.4, bothP< 0.01].Conclusion AGM can dramatically alleviate spleen immunosuppression, excessive inflammation and organ damage induced by trauma/hemorrhage.

OBJECTIVETo investigate the effects of hysteroscopic treatment of women with previous cesarean scar defect (PCSD).

METHODSFrom May 2006 to October 2008, 12 patients with PCSD were diagnosed and treated hysteroscopically in our hospital, all of them were successful followed-up for one year postoperatively, and their clinical data were analyzed.

RESULTSAll 12 hysteroscopic procedures were completed successfully, and there were no surgical complications. Nine patients with longer periods and 1 patient with intermenstrual spotting preoperatively remained asymptomatic after hysteroscopic surgery, and 1 patient with longer periods and infertility experienced normal periods, while remained infertility, and the remaining 1 patient complaining postcoital bleeding preoperatively had recurrence of the bleeding.

CONCLUSIONHysteroscopic surgery of women with PCSD was minimally invasive and effective.

Adult ; Cesarean Section ; adverse effects ; Cicatrix ; etiology ; surgery ; Female ; Follow-Up Studies ; Humans ; Hysteroscopy ; Postoperative Complications ; surgery ; Uterine Diseases ; surgery

This study is to investigate the anti-tumor effect in vitro of methotrexate modified by LH-RH peptide (LH-RH-MTX). LH-RH receptors highly expressing MCF-7 human breast carcinoma cell line and lowly expressing K562 human erythroleukemia cell line were served as the tested cells. The cell proliferation inhibition rates of LH-RH-MTX were detected by MTT colorimetric assay. The effects of LH-RH-MTX on the cell cycle and apoptosis rates were detected by flow cytometry. The inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that on K562 cells, and the inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that of free MTX at the same concentration. The inhibition rate of LH-RH-MTX on rat bone marrow mononuclear cells was less than that of free MTX. The number of MCF-7 cells in S phase increased after administration of LH-RH-MTX. The apoptosis rate of LH-RH-MTX group significantly increased compared with that of the control group and MTX group. The relative expression of LHRHR mRNA of LH-RH-MTX group markedly decreased compared with that of the control group and MTX group. LH-RH-MTX is realizable to reduce drug side effects, increase the therapeutic index and achieve tumor-targeted therapy.
Animals ; Antimetabolites, Antineoplastic ; chemical synthesis ; pharmacology ; Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drug Delivery Systems ; Gonadotropin-Releasing Hormone ; chemistry ; pharmacology ; Humans ; K562 Cells ; Leukocytes, Mononuclear ; MCF-7 Cells ; Methotrexate ; chemical synthesis ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Receptors, LHRH ; biosynthesis ; genetics

Erectile dysfunction (ED) is one of the common male diseases, which affects 15,200 men in the world. Recent discovery has shown the important role of reactive oxygen species (ROS) in ED. This paper updates the researches on the role of ROS in ED, expounds the relationship of ROS with various risk factors of ED, and outlines the advances in the anti-oxidation treatment of the problem.
Erectile Dysfunction ; etiology ; Humans ; Male ; Reactive Oxygen Species

OBJECTIVETo determine vacA dominant genotypes of Helicobacter pylori in patients with peptic ulcer (PU) or chronic gastritis (CG).

METHODSH.pylori strains were isolated from mucosa samples of gastric antrum and corpus of patients suffering from PU (n=29) or CG (n=34), 126 strains of H.pylori were selected for PCR to detect s and m regions in vacA gene of the isolates. Parts of the amplification products were sequenced after T A cloning. The correlation between infection or coinfection with different vacA genotypes of H.pylori and different gastroduodenal diseases was further analyzed.

RESULTSThe positive amplification products of vacA gene, s and m regions, were found in the DNA samples of all the isolates. In these products, s1a/m1, s1a/m2, s1a/m1b and s1a/m1b m2 genotypes of vacA gene were detected and s1b and s2m1a genotypes absent. Proportions of the s1a/m1, s1a/m2, s1a/m1b and s1a/m1b-m2 genotypes were 7.1% (9/126), 61.9% (78/126), 29.4% (37/126) and 1.6% (2/126), respectively. 17.5% (11/63) of the patients were confirmed to be coinfected with different genotype H.pylori strains. No statistical differences were found in the distribution of different genotype H.pylori strain infection in the gastric diseases (P>0.05). In comparison with the reported sequences of H.pylori strain 60190 with s1a genotype and strain 87-203 with m2 genotype, homologies of the nucleotide sequences of s1a PCR products from 6 strains of H.pylori isolates and m2 PCR products from 4 strains of H.pylori isolates were 93.15% approximate, equals 94.86%and 93.63% approximate, equals 97.61%, respectively.

CONCLUSIONH.pylori with s1a/m2 or s1a/m1b are the dominant genotypes in the PU or GC patients in Zhejiang area. The nucleotide sequences of partial amplification products from the vacA dominant genotypes of H.pylori show high homology compared with the reported sequences. Part of the patients may be coinfected with different vacA genotypes of H.pylori.

Adolescent ; Adult ; Aged ; Bacterial Proteins ; genetics ; Base Sequence ; Child ; Female ; Genotype ; Helicobacter pylori ; genetics ; Humans ; Male ; Middle Aged

To determine the optimum conditions of supercritical CO2 extraction of Xiaoyaosan, and establish its fingerprint by gas chromatography-mass spectrometry (GC-MS), the yield of extract were investigated, an orthogonal test was used to quantify the effects of extraction temperature, pressure, CO2 flow rate and time, and fingerprint analysis of different batches of extracts were by GC-MS. The optimal extraction conditions were determined as follows: extraction pressure 20 MPa, extraction temperature 50 degrees C, CO2 flow rate 25 kg x h(-1), extraction time 3 h, and average yield 2.2%. The GC-MS fingerprint was established and 27 common peaks were found, whose contents add up to 81.89% of the total peak area. Among them, 21 compounds were identified, accounting for 53.20% of the total extract. The extraction process is reasonable and favorable for industrial production. The GC-MS method is accurate, reliable, reproducible, and can be used for quality control of supercritical CO2 extract from Xiaoyaosan.
Carbon Dioxide ; chemistry ; Chromatography, Supercritical Fluid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods