In order to change the situations such as students’passivity and low enthusiasm in the traditional experimental teaching of Microbiology and improve students’comprehensive qualities,exploration and attempt on experimental teaching reforms including teaching contents,teaching modes,laboratory management and evaluation system of examination were done.Teaching practices showed that the normalized ex- periment syllabus based on the correct orientation of experimental teaching of Microbiology,the actualization of teaching mode of basic skill training-integrated experiment-designed experiment and the management of opening laboratory were effective measures in improving experimental teaching of Microbiology.

Objective To study the relationship between tyrosine phosphorylation(TP)and protein expression of insulin receptor substrate-1(IRS-1)and insulin resistance in patients with gestational diabetes mellitus(GDM).Methods IRS-1 expression and TP in skeleton muscle tissue were determined by Western blot and immunoprecipitation in women with GDM(GDM group,n=22),normal pregnant women(normal pregnancy group,n=22)and normal nonpregnant women(normal nonpregnant group,n=13).Fasting plasma glucose(FPG)and fasting insulin(FINS)were measured by oxidase assay and immunoradioassay. Results(1)The levels of FPG,FINS,and insulin resistance index were calculated according to homeostasis model assessment [ HOMA-IR;(5.6?0.8)mmol/L,(15.4?5.1)mU/L,and 1.2?0.5 ] in GDM group were significantly higher than those in normal pregnancy group [(4.4+0.5)mmol/L,(10.6 ?3.1)mU/L,and 0.8?0.3;P

Hemimasticatory spasm is a rare neurologic disorder characterized by unilateral, paroxysmal involuntary contraction of the masticatory muscles. It can be confused with other unilateral facial spasms. In this report, we present two patients with hemimasticatory spasm who were initially misdiagnosed with other disorders. The first patient was a 54-year-old man with a 9-year history of right facial spasm. He underwent microvascular decompression, without benefit, at another hospital with the diagnosis of hemifacial spasm two years prior to presenting at our hospital.On examination, right masseter and temporalis muscles had irregular contractions with twitches and prolonged spasms. His brain magnetic resonance imaging was normal.He showed a good response to carbamazepine. The second case was a 60-year-old man presented with a 10-year history of jaw-closing movement. Previous treatment for temporomandibular joint disorder was ineffective. He was previously diagnosed as oromandibular dystonia at another hospital, and biperiden and diazepam treatment resulted in slight improvement. There was tonic contraction and hypertrophy of the left masseter. He improved with carbamazepine. These cases illustrate the importance of hemimasticatory spasm as differential diagnosis, and the good response to carbamazepine.
Nervous System Diseases

Objective To investigate the effects of injecting allogeneic mesenchymal stem cells(MSCs) on the cellular immunity in rat in vivo.Methods Bone marrow-derived MSCs were isolated from Wistar rats.The purity of MSCs was identified by morphological examination with microphotography,and the phenotypes were identified with flow cytometry.Twenty SD rats were randomly divided into 4 groups.Different concentrations of MSCs(5?106/ml for group A,5?105/ml for group B,and 5?104/ml for group C,respectively) and PBS(for group D) were given to allogeneic SD rats via intravenous infusions.The suppressive effect of MSCs on lymphocytes proliferation in recipient rat was analyzed using mixed lymphocyte cultures(MLR) 10 days after cultivation.At the same time,proportions of CD4+,CD8+ and CD4+CD25+/CD4+ T-lymphocytes in peripheral blood and spleen were analyzed with flow cytometry.Results Proliferation rate of splenic lymphocytes in group A(5?106/ml MSCs,8.58%?0.27%) was markedly lowered compared with that in group D(PBS,24.40%?5.21%,P

BackgroundReactive oxygen intermediate products lead to the oxidative injury of cells.Retinal pigment epithelial(RPE) cells produce lots of reactive oxygen intermediate products during the swallow of out disc,but how this procedure cause the persistent oxidative injury of RPE cells is poorly understood.Objective The present study was to evaluate the effect of non-lethal H2 O2 -induced persistent oxidative injury on RPE barrier in vitro.MethodsARPE-19 cell links were inoculated on 96 well plate at the density of 8×104 cells/L and the cell climbing slice of 24 well at the density of 4× 104 cells/L.The cells were cultured in DMEM/F12 medium,and the cells cultured for 24 hours in free-serum medium were used in the experiment.0-0.6 mmol/L of H2O2 were added into the medium.Cellular viability was assessed using 3- ( 4,5-dimethylthiazol-2-yl ) -5- ( 3-carboxymethoxyphenyl ) -2- ( 4-sulfophenyl ) 2H-tetrazolium(MTS) assays.Transepithelial electrical resistance (TER) was used to detect cell monolayer forming time after cultureinTrsnswellchamber.Thepermeabilityof cellmonolayer was examinedbyrhodamine isothiocyanate-dextran transepithelial flux,and immunofluorescence was used to investigate the distribution of the junction protein zonula occludens (ZO-1).ResultsThe total difference was found in the cell vitality(A490) among the different concentrations of H2 O2 ( F =991.501,P =0.000 ).Compared with 0 mmoL/L H2 O2 group,the A490 values was gradually lowed from 0.20 mmol/L H2O2 group to 0.60 mmol/L H2O2 group (P ＜ 0.05 ).H2O2 at the concentrations of ＞0.20 mmol/L lowed the viability of RPE cells.The TER value was ( 24.9 ± 1.3 ) Ω · cm2 in 11 days,( 17.8± 1.4)Ω · cm2 in 7 days after inoculation on transwell chamber,showing a significant difference between them (t=5.228,P=0.014).RPE formed the stable tight junction on day 15 with the TER value (25.9±0.9 ) Ω · cm2.The leakage amount ( relative fluorescence intensity ) of the dextran was 255.39 ± 16.44 in non-H2 O2 control group,exhibiting a significant lowing in comparison with free-cell blank group (433.08±51.53)( t =12.515,P =0.006 ),and that of H2 O2 group was significant increased in comparison with non-H2 O2 control group ( t =14.412,P=0.005).Immunofluyorescence assay showed intact intercellular ZO-1 junction in non-H2O2 control group,but the breakage of ZO-1 junction was seen in H2O2 group.ConclusionsThe results indicate that non-lethal H2O2 can destroy RPE barrier and further lead to the persistent oxidative injury of RPE cells.

Baculovirus DNA polymerase gene belongs to an early gene of baculovirus. It is a necessary gene required for replication of virus in insect cells. It can encode DNA polymerase induced by virus. In the process of replication, DNA polymerase can bind to homologous regions and non-homologous regions, which are believed to act as the origins of virus DNA replication with other replication factors. In addition, DNA polymerase has advantages over occlusion protein and egt gene for resolving deep branching taxonomic relationships of baculovirus phylogenies.

Objective To report the development of clinical symptoms in a Chinese family with autosomal dominant progressive external ophthalmoplegia(adPEO).Methods Electromyologram and muscle biopsy were performed in the proband and 4 family members with the disease.Results The proband was a 57 year-old woman,who developed bilateral ptosis after the age of 30,external ophthalmoplegia after the age of 35 years old,weakness of extremities at the age of 37 years old and bulb palsy with palmus at the age of 47 years old.In the family there were 20 male and female members from five generations.All of them complained about bilateral ptosis between 26—33 years old,external ophthalmoplegia(12/15)and weakness of all extremities(14/15)between 35—45,facial and masticatory weakness(9/9)as well as dysphagia(8/9)between 44—60,accompanied with heart lesions(4/7)after 50 years old.Some patients died due to cardiac impairment.Electromyologram showed myopathic abnormalities in the examined patients. The main myopathological changes were ragged red fibers,cytochrome c oxidase negative fibers and ragged blue fibers in succinate dehydrogenase staining.Conclusions The adPEO started from extra-ocular muscles to limbs,finally facial and bulbar muscles.Heart lesions were presented in late stage and lead to death in some members.The developing process of symptoms suggested that we should pay more attention to cardiac manifestations in this disease.

OBJECTIVETo observe the clinical efficacy of treating chronic persistent bronchial asthma (CPBA) children with abnormal myocardial enzyme spectrum (AMES) by Yupingfeng Powder (YP) combined routine therapy.

METHODSFrom January 2010 to December 2012, 156 CPBA children patients with AMES were randomly assigned to the treatment group (80 cases) and the control group (76 cases). All patients received routine treatment (inhaled corticosteroids and/or leukotriene regulator). Besides, those in the treatment group took YP. The treatment duration was 3 months. The scores of children asthma control test (C-ACT), pulmonary function (FEV,% and PEF%), myocardial enzyme spectrum were observed before and after treatment, and 3 months before and after treatment. The myocardial enzyme spectrum of 40 healthy children at the baby clinics during the same period were recruited as the control.

RESULTSCompared with the control group, creatine kinase isoenzyme (CK-MB), creatine kinase(CK), and lactate dehydrogenase (LDH) increased in the two treatment groups (P <0.01), but there was no statistical difference in AST (P >0.05). Compared with before treatment in the same group, CK-MB, CK, LDH, and AST decreased in the treatment group after treatment and 3 months after treatment (P <0.01). CK-MB, CK, LDH, and AST decreased in the control group 3 months after treatment (P <0.01, P <0.05).Compared with after treatment, CK decreased in the control group 3 months after treatment (P <0.01). C-ACT score, FEV(1),%, and PEF% all increased in the two groups after treatment and 3 months after treatment (P <0.01, P <0.05). Compared with after treatment in the same group, CK decreased in the control group 3 months after treatment (P <0. 01). Compared with the control group in the same period, post-treatment CK-MB and CK decreased (P <0. 01, P <0. 05), while post-treatment C-ACT score, FEV, %, and PEF% increased (P <0.05) in the treatment group (P <0.05).

CONCLUSIONYP could strengthen specific and non-specific immunity of the organism, and improve clinical symptoms and the level of myocardial enzyme spectrum.

Asthma ; therapy ; Child ; Chronic Disease ; therapy ; Creatine Kinase, MB Form ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Myocardium ; enzymology

OBJECTIVETo explore the effect of qidong huoxue decoction (QHD) on inflammatory factors and Toll-like receptor (TLR4) mRNA expressions in acute lung injury (ALI) rats.

METHODSTotally 50 healthy male SD rats were randomly divided into the blank control group, the lipopolysaccharide (LPS) model group, low, middle, high dose QHD groups according to body weight, 10 rats in each group. Rats in low, middle, high dose QHD groups were intragastrically administered with QHD at 4, 8, and 16 mL/kg 24, 12 h before modeling and 12 h after modeling, respectively. Normal saline was intragastrically administered to rats in the blank control group and the LPS model group. An ALI rat model was established using intratracheal instillation of LPS. Rats were killed after 24-h modeling. Then the bronchoalveolar lavage fluid was prepared. Contents of TNF-α, IL-1β, and L-10 were detected using ELISA. TLR4 mRNA expressions were determined byreal time PCR.

RESULTSCompared with the blank control group, contents of TNF-α, IL-1β , and IL-10 increased (P <0. 01), TLR4 mRNA expressions also increased in the LPS model group (all P <0. 01). Compared with the LPS model group, contents of TNF-α and IL-1β decreased (P <0. 05, P <0. 01), IL-10 levels increased (P <0. 01) , TLR4 mRNA expressions were also reduced (P <0. 01), in high and middle dose QHD groups. Compared with the high dose QHD group, con- tents of TNF-α and IL-1β increased in middle and low dose QHD groups (P <0. 05); IL-10 levels decreased (P <0. 05) in the low dose QHD group(P <0. 05), TLR4 mRNA expressions also increased in the low dose QHD group (P <0. 05). Compared with the middle dose QHD group, IL-10 levels was reduced, but TLR4 mRNA expressions increased in the low dose QHD group (P <0. 05).

CONCLUSIONSQHD had the protective effect on LPS induced ALI rats. Its mechanism might be associated with inhibiting TLR4 mRNA expressions, leading to decreased pro-inflammatory cytokines such as TNF-α and IL-β, elevated anti-inflammatory cytokine IL-10, and thereby, correcting unbalanced inflammation.

Acute Lung Injury ; genetics ; metabolism ; Animals ; Anti-Inflammatory Agents ; Bronchoalveolar Lavage Fluid ; Drugs, Chinese Herbal ; pharmacology ; Inflammation ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; Male ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

AIM: To establish an experimental model of high intraocular pressure in mice by laser photocoagulation and to prepare for future research.
METHODS: Experimental model of high intraocular pressure was induced unilaterally in 44 C57BL/6 mice. The fellow eye served as a control. TONO-PEN AVIA Tonomter was used to measure intraocular pressure (IOP) to guarantee IOP value at 1, 2, 4, 8wk. Slit-lamp biomicroscopy was performed throughout the period and the structural changes were assessed histologically. And then, their eyes were enucleated, postfixed, cryoprotected, and embedded in optimal cutting temperature medium. After hematoxylin and eosin stain ( HE stain ) , cryosections of the retina were observed under light microscope. TdT-mediated biotin-dUTP nick end labeling ( TUNEL ) was performed on the retinal sections to determine apoptosis rate.
RESULTS: IOP of laser-treated eyes was significantly higher than that of control eyes from 1-8wk (P<0. 05). The highest IOP was 31mmHg, but only one eye. The IOP was mainly around 20mmHg. In laser-treated eyes, the angle of anterior chamber were narrow. Number of cells in the inner nuclear layer and retial gangllion cell layer was slightly lower than that in control eyes at 2wk, but by 4 and 8wk the number of cells was significantly lower than that in the control contralateral eyes.
CONCLUSION: The laser photocoagulation of limbus causes chronic elevation of IOP and this method may be a promising experimental model for the investigation of biological mechanisms of glaucomatous retinal ganglion cell damage.