AIM: To construct a high-level expression system of recombinant human neuronal nitric oxide synthase (hnNOS) full-length enzyme in Escherichia coli. METHODS: The coding sequence of hnNOS full-length was firstly amplified by PCR, and then ligated into the expression vector pCWori+. The recombinant plasmid was transformed into Escherichia coli BL21 for high-level expression. After having been checked with Western blot, the enzyme was used for large-scale culture and purification. Finally, the property of the enzyme was determined by spectrophotometric method. RESULTS: The constructed expression system could give a yielding of 3 mg/L initial culture. CONCLUSION: The expression system constructed is fully sufficient to express the active human neuronal nitric oxide synthase.

Myasthenia gravis (MG) is an immune-mediated disorder of the neuromuscular junction that is characterized by fluctuating muscle weakness.Acetylcholine receptor antibody (AchR-Ab) is the first MG specific antibody being elucidated,however,the discovery of many other MG specific or related antibodies enriches its immunopathogenesis greatly.The detection of antibodies is very important for the diagnosis and classification of MG.It can be used to guide the clinical management and evaluate the response to treatment as well.With the development of laboratory medicine,the methods of antibody detection are also being optimized.In this paper,we will review the serum antibodies closely associated with MG and their mechanisms,detection methods and clinical significance.

Objective To investigate the effect of continuous blood purification (CBP) on pro-or anti-inflammatory immune function and prognosis in patients with sepsis. Methods One hundred and two patients with sepsis were randomized into the CBP group (60 cases) and the control group (42 cases). The patients in the control group were treated with conventional therapy, and the patients in the CBP group received at least 72 h CBP treatment additionally. The APACHEⅡ score, SOFA score, the 28 day survival rate and ICU length of stay were recorded and levels of spleen Th1, Th2 were assessed by FACS flow cytometry. Enzyme linked immunoadsordent assay (ELISA) was used to determine the levels of serum IL-1, IL-10 and TNF-α before and at 24, 48, 72 h after the treatment. Results The APACHEⅡscore and SOFA score decreased markedly in the CBP group after the treatment (Р <0.05). The period of staying in ICU of patients in the CBP group was shorter than that of patients in the control group (Р < 0.05). There was no significant difference of the 28 day survival rate between the two groups (91.6% vs 71.2%, Р > 0.05). Compared with the control group, levels of IL-1, IL-10 and TNF-α were decreased markedly, and the ratio of Th1 / Th2 was increased significantly at 72 h after the treatment in the CBP group (Р < 0.05). Conclusion CBP can eliminate inflammatory mediators, and help to enhance the immune function, and restore the balance of Th1 / Th2 in patients with severe sepsis.

Objective To investigate the safety and effectiveness of Glifeet in bowel preparation before colonoscopy.Methods A prospective,single blind,randomized controlled trial of patients undergoing colonos-copy was conducted.A total of 67 inpatients were randomized to the control group,who received a low-residue and semiliquid diet (n =31)and the experimental group,who received Glifeet all day (n =36)before the proce-dure.All patients took polyethylene glycol electrolyte powder 1 000 ml at 7 pm on the day before colonoscopy and 2 000 ml at 8 am on the examination day.Bowel preparation quality was scored using the Boston Bowel Prepara-tion Scale (BBPS).Side effects were also observed.Results The time of the first bowel movement of the experi-mental group was significantly shorter [(77.43 ±54.21)min VS (149.35 ±118.15)min,P =0.002].An in-creased defecating frequency was observed in the experimental group,but there was no significant difference when compared with the control group (11.44 ±6.95 VS 8.74 ±3.58,P =0.055).Patient tolerance and acceptance did not differ.There was no significant difference in BBPS between the two groups.But the bowel preparation quality of the right colon was significantly better in the experimental group (2.56 ±0.50 VS 2.23 ±0.81,P =0.045). Conclusion Glifeet could meet the needs of basic energy in most patients for colonoscopy.Furthermore,Glifeet is well tolerated and can partially improve the quality of bowel preparation.

Objective To investigate the inhibitory effect of the cytochrome P450 3A4(CYP3A4) gene expression and function in CHL-3A4 cells lines by vector-ecpressed small hairpin interfering RNA(shRNA).Methods The shRNA expression vectors targeting CYP 3A4 gene(CYP3A4Ⅰ,CYP3A4Ⅱ,CYP3A4Ⅲ) were designed and constructed.The inhibitory effect of shRNA was detected by Western blot and RT-PCR analysis.The inhibitory effect of cytotoxic of cyclophosphamide using shRNA was measured by MTT assay.Results CYP3A4Ⅲ shRNA expressing vector significantly reduced the CYP3A4 mRNA(70%) and protein expression levels(75%) of the CYP3A4 gene in CHL3A4 cells suppression of CYP3A4 gene expression by CYP3A4Ⅲ largely reversed cyclophosphamide cytotoxicity(75%) in CHL-3A4 cells.Conclusion Vector-based RNAi could suppress CYP3A4 gene expression and function,and the use of RNAi to inhibit CYP3A4 gene expression in mammalian cells was a promising new tool for the study of cytochrome P450 gene function.

[ ABSTRACT] AIM:To investigate the role of hydrogen sulfide ( H2 S) on impaired wound healing in ob/ob mice and the underlying mechanism.METHODS:The ob/ob mice were randomly divided into 3 groups, including vehicle, in-sulin and NaHS for treatment.C57BL/6 mice were treated with vehicle as control.Full-thickness punch biopsy wounds were created on the mice.Firstly, H2 S concentrations in the skins and granulation tissues were measured.The mRNA ex-pression of cystathionineγ-lyase ( CSE) was detected by RT-qPCR.The protein expression of CSE and MMP-9 were deter-mined by Western blot.The neutrophil and monocyte/macrophage infiltration was analyzed by immunohistochemistry me-thod.The levels of tumor necrosis factor (TNF)-αand interleukin (IL)-6 were measured by ELISA.Collagen formation was measured by Masson staining.RESULTS:The H2 S levels in the skin and granulation were significantly decreased in ob/ob mice and increased in the NaHS-treated mice ( P<0.05 ) .CSE expression at mRNA and protein levels was significantly decreased in ob/ob mice compared with the control mice (P<0.05).The wound healing period was significantly shorter in NaHS group than that in vehicle-treated ob/ob mice group (P<0.05), in which the insulin group had no difference with vehicle ob/ob mice group.The neutrophil and monocyte/macrophage infiltration, and TNF-αand IL-6 levels were signifi-cantly increased in ob/ob groups, but were decreased in NaHS group ( P<0.01 or P<0.05 ) .Meanwhile, NaHS in-creased collagen formation in the granulation tissues of ob/ob mice.CONCLUSION:H2 S/CSE down-regulation contributes
to impaired wound healing in diabetes, which is alleviated by exogenous H2 S possibly through anti-inflammation.

Flow cytometry (FCM) is a technology for fast qualitation, quantitative analysis and separation of single cells or other biological particles. The method is rapid, sensitive and accurate, and its objective and direct result can be analyzed with multi-parameters simultaneously. The morphological, biochemical and molecular changes during apoptosis include cellular shrinkage, permeability transition of plasma membrane, caspases activation, dissipation of mitochondrial trans-membrane potential, phosphatidylserine redistribution, calcium flux and DNA fragmentation and content. The review outlined the methods to characterize, identify and quantify apoptoUc cells by flow cytometry to further determine cell apoptosis.

Objective To evaluate the effects of a health education and behavior intervention project,and to provide.a scientific basis for brucellosis control.Methods By stratified cluster sampling method,target population from 6 township in Meilisi District were randomly selected to carry out baseline survey.Of these 6 townships 4 were selected as intervention townships,and two as control townships.A health education and behavior intervention program was carried out in intervention townships,and after intervention target population from 6 township were participated in the questionnaire survey.Results Awareness rate of common sense knowledge of farmers in intervention townships was 84.33％ (5237/6210) after intervention which was significantly higher than 53.56％ (2003/3740),before the intervention (x2 =112.49,P ＜ 0.01),and also higher than 54.15 ％ (1787/3300) of the control after the intervention (x2 =101.53,P ＜ 0.01).Average awareness rate(82.13％,2550/3105) of common sense on prevention after intervention was significantly higher than 58.77％(1099/1870) before the intervention (x2 =57.19,P ＜ 0.01).After intervention,average awareness rate of common sense on prevention was significantly higher than that of the average of control townships (70.36％,1161/1650,x2 =25.49,P ＜ 0.01).The awareness rate of treatment before intervention was 36.80％ (116/299),which increased to 76.36％ (436/571)after intervention (x2 =119.38,P ＜ 0.01).After the intervention the awareness rate of intervention townships was significantly higher than that of the controls (33.72％,88/261,x2 =139.69,P ＜ 0.01).In survey of these high-risk behavior in intervention townships,in addition to cattle and sheep which were regularly immunized in the intervention townships(35.91％,121/337),and the control townships(32.01％,97/303,x2 =1.08,P ＞ 0.05) and their difference was not significant,other processing such asapoblema,killing cattle,sheep,cattle to deliver,cutting the wool,regular disinfection,and accuracy of high-risk behavior protection were significantly higher than those of controls[89.83％(106/l18),91.07％(51/56),84.75％(150/177),66.67％(32/48),73.78％(242/328),51.22％(42/82),75.56％(34/45),33.69％(63/187),27.78％(15/54),21.63％(61/282),x2 value were 38.00,5.82,103.84,15.84 and 173.67,all P ＜ 0.05].Conclusion After implementation of health education and behavioral intervention,farmers brucellosis prevention knowledge and awareness of risk behaviors have significantly improved,which have achieved the desired effect.

OBJECTIVE: To observe the effect and explore the mechanism of Huannao Yicong capsule in treating senile patients with mild cognitive impairment (MCI). METHODS: The investigational drugs were packed by blind method. A randomized, double-blind and controlled trial was conducted on ninety senile patients with MCI. Other forty-five senile healthy persons were recruited to the healthy control group. The ninety senile patients were randomly divided into the Huannao Yicong capsule-treated group (45 patients administered with three Huannao Yicong capsules and two aniracetam capsule analogues) and aniracetam-treated group (45 patients treated with two aniracetam capsules and three Huannao Yicong capsule analogues). Patients in the two groups were treated three times daily for 16 weeks. Memory, traditional Chinese medicine syndrome, cerebral blood flow, free radicals and inflammatory mediators, such as superoxide dismutase (SOD), malondialdehyde (MDA), acetylcholinesterase (AchE), interleukin-1alpha (IL-1alpha) and interleukin-6 (IL-6) were determined before and after the treatment. Blood lipids, including triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), apolipoprotein A-1 (ApoA-1) and apolipoprotein B-100 (ApoB-100), were detected before and after the treatment. The safety indexes, such as routine tests of blood and urine, hepatic and renal function tests and electrocardiogram (ECG) were taken before and after the treatment. RESULTS: Index score of clinical memory scale in senile healthy people was significantly higher than that in MCI patients before treatment (P<0.01), and the content of AchE, IL-1alpha and IL-6 was obviously lower (P<0.01, P<0.05), the activity of SOD was higher (P<0.05). No significant difference was found in direction memory of clinical memory scale between the two treatment groups. Other index scores of clinical memory scale and traditional Chinese medicine syndrome in patients of Huannao Yicong capsule-treated group were significantly improved as compared with those of the aniracetam-treated group (P<0.05, P<0.01). The blood flow parameters of anterior cerebral artery, posterior cerebral artery and resistant index in patients of Huannao Yicong capsule-treated group were increased significantly (P<0.01, P<0.05). Huannao Yicong capsule could significantly increase the activity of serum SOD and decrease the content of AchE, IL-1alpha and IL-6 (P<0.01, P<0.05), better than aniracetam. Furthermore, Huannao Yicong capsule could significantly improve the blood lipid, such as the level of TG, LDL-C, HDL-C, ApoA-1 and ApoB-100 (P<0.01, P<0.05), and better than aniracetam (P<0.01, P<0.05). No significant changes were found after treatment in safety indexes, such as routine tests of blood and urine, hepatic and renal function tests and ECG. CONCLUSION: Huannao Yicong capsule has better therapeutic effect than aniracetam capsule in treating senile mild cognitive impairment.

PurposeTo discuss the imaging features of multi-slice spiral CT (MSCT) and PET/CT in lymphoma with pulmonary lesions as the initial presentation.Materials and Methods Eighteen cases of lymphoma with pulmonary lesions as initial presentation confirmed by operation and pathology were retrospectively analyzed including Hodgkin lymphoma (HL) in 3 cases and non-Hodgkin lymphoma (NHL) in 15 cases. All 18 patients underwent MSCT, of which 11 also underwent PET/CT. The MSCT manifestations included number of lesions, morphology, density and bronchial change. PET/CT evaluated lesions in other body parts and SUVmax of lung lesions.Results MSCT showed single lung lesion in 6 cases, multiple lung lesions in 12 cases. Masses were seen in 9 cases, non-masses in 7 cases, and mixed type in 2 cases. There was single consolidation in 3 cases and mixed consolidation in 15 cases. Air bronchogram was identified in 15 cases. CT predictive diagnosis was lymphoma in 3 cases, lung cancer in 5 cases, pneumonia in 7 cases, granuloma in 1 patient, and indeterminate in 2 cases. PET/CT showed hypermetabolic lung lesions in 11 patients, mediastinal lymph node in 3 cases, and unilateral supraclavicular lymphadenopathy in 1 case. One patient showed pancreatic tail masses, gastric wall thickening and haunch masses. All 11 patients were diagnosed as malignant tumor, of which 8 cases as lymphoma.Conclusion MSCT manifestations of lymphoma with pulmonary lesions as initial presentation include multiple lesions, polymorphic type and heterogeneity with air bronchogram. PET/CT has better accuracy than MSCT.