Objective To investigate the effects of zinc deficiency on the expressions of Th1/Th2 cytokine interferon-?(IFN-?) and interleukin-4(IL-4)in asthmatic rats.Methods Animal models of asthma and zinc deficiency were established.Thirty-two SD rats were divided into 4 groups according to weight:zinc deficient diet with ovalbumin(OVA) challenge group(group A),zinc normal diet with OVA challenge group(group B),zinc normal pair-fed diet with OVA challenge group(group C) and zinc normal diet with saline challenge group(group D).The contents of IFN-? and IL-4 in lung homogenate were detected by enzyme linked immunosorbent assay(ELISA).The levels of IFN-? and IL-4 mRNAs were observed by reverse transcriptase-polymerase chain reaction(RT-PCR).Results Compared with group D,the content and mRNA expression of IFN-? of lung in group A,group B and group C all decreased significantly(all P0.05).No marked difference was found between group B and group C in all the results.Conclusions Zinc deficiency reduces the expression of IFN-? but have no influence on IL-4.The imbalance of Th1/Th2 cytokine may be associated with the increasing airway inflammatory reaction in zinc deficient asthmatic rats.

OBJECTIVETo explore the intervention of baicalin on signal transduction and activating transcription factor expression of ulcerative colitis (UC) patients.

METHODSRecruited were UC patients at Outpatient Department of Digestive Disease, Inpatient Department of Digestive Disease, Center for Digestive Endoscopy of College City Branch, Guangdong Provincial Hospital of Traditional Chinese Medicine, and Southern Hospital affiliated to Southern Medical University from June 2010 to January 2011. They were assigned to the UC group (33 cases) and the diarrhea-predominant irritable bowel syndrome (IBS-D) group (30 cases). Another 30 healthy subjects were recruited as a healthy control group. Peripheral blood mononuclear cells (PBMCs) in vitro intervened by different concentrations baicalin were taken from UC patients. IL23R gene expressions in vitro intervened by different concentrations baicalin were detected using Q-PCR. Expressions of signal transducer and activator of transcription 4 (STAT4) , STAT6, phosphorylated-STAT4 (p-STAT4), and p-STAT6 were detected using Western blot. Serum levels of IFN-γ, IL-4, IL-6, and IL-10 were measured by ELISA. Effects of different concentrations baicalin on expressions of PBMCs, and levels of IFN-γ, IL-4, IL-10 of UC patients were also detected.

RESULTSCompared with the negative control group, 40 µmol baicalin obviously decreased IL23R gene expression of UC patients (P <0. 01). Compared with the healthy control group and the IBS-D group, p-STAT4/STAT4 ratios increased, p-STAT6/STAT6 ratios decreased, levels of IFN-γ, IL-4, IL-10 all increased in the US group (all P <0. 05). Compared with the negative control, 5 and 10 µmol baicalin groups, 20 and 40 moL baicalin obviously decreased p-STAT4/STAT4 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously increased p-STAT6/STAT6 ratios (all P <0. 05); 20 and 40 µmoL baicalin obviously lowered levels of IFN-γ and IL-4, and elevated IL-10 levels (all P <0. 05).

CONCLUSION40 µmoL baicalin could in vitro inhibit p-STAT4/STAT4 ratios, adjust p-STAT6/STAT6 ratios and related cytokines, thereby balancing the immunity and relieving inflammatory reactions of UC.

Activating Transcription Factors ; metabolism ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Blotting, Western ; Colitis, Ulcerative ; drug therapy ; metabolism ; Cytokines ; metabolism ; Flavonoids ; therapeutic use ; Humans ; Interleukin-10 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Irritable Bowel Syndrome ; drug therapy ; metabolism ; Leukocytes, Mononuclear ; Medicine, Chinese Traditional ; Phosphorylation ; STAT6 Transcription Factor ; metabolism ; Signal Transduction

OBJECTIVETo construct plant expression pCAMBIA1301-PMI by substituting PMI for hygromycin resistance gene in pCAMBIA1301 and obtain transgenic Salvia miltiorrhiza f. alba using PMI-mannose selection system.

METHODThe 6-phosphomannose isomerase gene (PMI) of Escherichia coli was amplified by PCR. Sequence analysis showed that it shared 100% amino acids identities with the sequences of PMI genes isolates reported in the NCBI. Based on pCAMBIA1305, the plant expression pCAMBIA1305-PMI was constructed successfully by substituting PMI for hygromycin resistance gene in pCAMBIA1305. pCAMBIA1305-PMI was transformed into Agrobacterium tumefaciens LBA4404, and then the leaves of S. miltiorrhiza f. alba were inoculated in LBA4404 with pCAMBIA1305-PMI.

RESULTPlant expression pCAMBIA1301-PMI was successfully constructed and the leaves of S. miltiorrhiza f. alba inoculated in LBA4404 with pCAMBIA1305-PMI were selected on medium supplemented with a combination of 20 g x L(-1) mannose and 10 g x L(-1) sucrose as a carbon source. The transformation efficiency rate was 23.7%.

CONCLUSIONGenetic transformation was confirmed by PCR, indicating that a new method for obtaining transgenic S. miltiorrhiza f. alba plants was developed using PMI-mannose selection system.

Anti-Bacterial Agents ; pharmacology ; Biomarkers ; Cinnamates ; pharmacology ; Escherichia coli ; enzymology ; genetics ; Escherichia coli Proteins ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hygromycin B ; analogs & derivatives ; pharmacology ; Mannose-6-Phosphate Isomerase ; genetics ; metabolism ; Plants, Genetically Modified ; drug effects ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; metabolism ; Transformation, Genetic

The fibrous roots are the residues of production of cut crude drug of Danshen (Salvia miltiorrhiza). Enzymatic pretreatment and ultrasonic extraction are beneficial to extract effective constituents from fibrous roots more effectively. The present research was to optimize the enzymatic parameters by the central composite design and response surface method. Under the best conditions, the yields of total tanshinones and total salvianolic acids in the extracts of enzymatic pretreatment increased by 113.92% and 30.64%, comparing with the non-enzymatic extraction, respectively. TLC analysis also showed that the types of effective constituents in the two samples were not affected by enzymatic hydrolysis. Meanwhile, the complex correlation coefficients of the mathematical models were high, which provided a good prediction.
Diterpenes, Abietane ; Drugs, Chinese Herbal ; chemistry ; Phenanthrenes ; chemistry ; Plant Roots ; chemistry ; Salvia miltiorrhiza ; chemistry

Objective To observe the association between adiponectin and small dense low-density lipoprotein (sLDL-c) in coronary artery disease (CAD) patients.Furthermore,we sought to determine the association between single nucleotide polymorphisms (SNP) rs1501299 ( + 276G/T ), rs266729 (-11365C/G) and the incidence of CAD.Methods Consecutive subjects with chest discomfort were examined by coronary angiography and divided into non-CAD [ n =250,147 male,mean age (60.26 ±7.52) years] and CAD [n =267,153 male,mean age (60.79 ±9.63) years] groups.Blood samples were collected from all participants following an overnight fasting for at least 12 hours.Plasma adiponectin levels were measured by competitive enzyme-linked immunosorbent assay (ELISA).The serum levels of sLDL-C and oxidized low-density lipoprotein (ox-LDL) were determined by ELISA.Genotypes in rs1501299 and rs266729 of the adiponeetin were determined by polymerase chain reaetion ( PCR ).Results 1.The adiponectin levels were significantly lower [ ( 306.17 ± 74.52 ) mg/L vs.( 321.78 ± 86.28 ) mg/L ],whereas sLDL-C and ox-LDL levels were significantly higher [ ( 276.30 ± 45.55 ) ng/L vs.( 249.00 ±32.02) ng/L and (545.06 ± 115.46 ) μg/L vs.(497.74 ± 106.09 ) μg/L,P ＜ 0.05 ] in CAD group than non-CAD group.2.Adiponectin level was negatively associated with sLDL-C,whereas sLDL-C positively correlated with ox-LDL in all subjects.3.Genotype distribution and allele frequencies of rs1501299 and rs266729 were similar between CAD and non-CAD subjects and not related to the serum levels of adiponectin, sLDL-C and ox-LDL.Conclusions Reduced adiponectin and increased sLDL-C were independent risk factors for coronary artery disease.Genetic polymorphisms in rs1501299 and rs266729 were not linked with coronary artery disease.

OBJECTIVETo explore the feasibility and effect of transumbilical single-site laparoscopic surgery in the treatment of inguinal cryptorchidism in children.

METHODSFrom August to November 2013, 33 children with inguinal cryptorchidism (41 testes) underwent transumbilical single-site laparoscopic orchiopexy. The undescended testes were palpable in the inguen intra-operatively in all the cases, 14 on the right, 11 on the left, and 8 bilaterally.

RESULTSAll the operations were performed successfully with neither intraoperative complications nor conversion to operi surgery. Adequate length of spermatic cord was pulled down to allow the testis to descend through the inguinal canal into the scrotum in all the cases. Totally, 39 testes in 31 cases were fixed at the bottom and 2 testes in 2 cases in the middle of the scrotum. Follow-up ranged from 6 to 9 months, which showed normal development of the testes, but no such postoperative complications as testicular retraction and atrophy, indirect hernia, and hydrocele.

CONCLUSIONTransumbilical single-site laparoscopic orchiopexy is a feasible and effective technique for the treatment of inguinal palpable undescended testis in children, and its cosmetic results were desirable.

Adolescent ; Child ; Child, Preschool ; Cryptorchidism ; surgery ; Feasibility Studies ; Humans ; Infant ; Laparoscopy ; methods ; Male ; Orchiopexy ; adverse effects ; methods ; Postoperative Complications ; Scrotum ; Spermatic Cord ; Testicular Hydrocele ; etiology

BACKGROUNDHepatitis C virus (HCV) infection may induce autoimmune response and autoantibodies can be detected in chronic hepatitis C (CHC) patients. However, the reported positive rate of autoantibodies in CHC patients in China varies considerably. In this study, we investigated the prevalence of antinuclear antibodies (ANA) and anti-liver-kidney-microsome type 1 autoantibodies (anti-LKM-1) in a large cohort of CHC patients, and analyzed the factors related to the presence of the autoantibodies.

METHODSA total of 360 CHC patients were enrolled in this study. Serum ANA and anti-LKM-1 were detected by indirect immunofluorescence and enzyme-linked immunosorbent assay, respectively. Clinical analysis was performed to disclose the related factors to autoantibody production.

RESULTSThe prevalence of ANA and anti-LKM-1 in CHC patients was 12.5% (45/360) and 2.5% (9/360), respectively. Women had a higher prevalence than men (18.9% vs 11.4%, P = 0.046). Patients with positive autoantibodies had lower HCV RNA levels (1.2 x 10(7) copies/L vs 7.2 x 10(7) copies/L, P < 0.05). Positive ANA was associated with higher serum globulin (P < 0.05). Stratified analysis showed that there were no significant differences in age, HCV genotype, disease course, clinical stage, prevalence of cirrhosis and interferon therapy between autoantibody-positive and -negative subgroups.

CONCLUSIONAutoantibodies can be induced in the course of CHC, and some CHC patients can even develop autoimmune hepatitis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Antinuclear ; blood ; Autoantibodies ; blood ; Child ; Child, Preschool ; China ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique, Indirect ; Hepatitis C, Chronic ; blood ; immunology ; Humans ; Male ; Middle Aged ; Prevalence ; Young Adult

Objective To observe the tropism ofboue marrow stromal stem cells for malignantglioma in rats. Methods The immunophenotype of in vitro cultureA Fisher344 rat BMSCs wereidentified using flow cytometry. The BMSCs or NIH3T3 cells were cocultured with 9L glioma cells in aTranswell system, and 24 h later, the cell migration rate was calculated. For in vivo experiment, aFisber344 rat model bearing malignant glioma was established by stereotactic injection of 9L glioma cellsinto the brain. After validation of the model 2 weeks after the injection by neurobehavioral test, magneticresonance imaging and HE staining, the BMSCs or NIH3T3 cells were transplanted via the internalcarotid artery in the rats. Two weeks after the transplantation, the rats were sacrificed by routine cardiacperfusion, and BMSCs migration in the brain was detected immunohistochemically. Results Thethird to six passages of the BMSCs were negative for CD34 and CD45 but positive for CD29 and CD44.Transwell assay demonstrated BMSCs tropism for 9L cells in vitro. In Fisher344 rats bearing 9L glioma,neurobehavioral changes characteristic of glioma were observed, and the BMSCs transplanted via theinternal carotid artery were found to migrate to the glioma tissue, residing mostly on the boundarybetween the normal tissue and the tumor tissue. Conclusion Rat BMSCs show a tropism formalignant glioma both in vitro and in vivo, and administration via the internal carotid artery can be aneasy and effective means for BMSCs transplant.

Objective To map the vision cortex of rats by dynamic manganese-enhanced functional magnetic resonance imaging and provide a method for researching the nervous function. Methods Six adult male Wistar rats were chosen and the process was divided into 4 continuous phases. No agent was injected into the rats in the first phase (5 min). Disrupting the BBB with marmitol and injecting manganese chloride were performed in the fight internal carotid artery (ICA) in the second phase (10 min). In the third phase (15 min), manganese chloride was administrated into theright ICA and vision stimulation was performed before the imaging process. The mixed liquor of manganese chloride and glutamate was injected into the rats in the forth phase (5 min). MRI was performed instantly after the handles in each phase. SPM and Matlab software were employed to help analyze the imaging data. Region-of-interest (ROI) was recorded to observe the stimulated regions and compare the signal intensity in the visual cortex. Results No specific enhanced region was found in the rat brain in the first and second phases. The right visual cortex was enhanced specifically on T1WI in the third phase. Many brain regions of the right hemisphere, the sites that agents was injected, were obviously enhanced in the forth 2008A1-E4011)phase. ROI analysis showed that the signal intensity in the third phrase (1.897±0.172) was significantly stronger as compared with that in the second phrase(1.549±0.163)(P<0.05). Conclusion The dynamic manganese-enhanced functional magnetic resonance imaging can analyze the functional activities of the vision cortex in rats and provide a new method for researching the function of the nervous system.

Objective To evaluate the efficacy and safety of oxiracetam in the treatment of neurological deficits resulting from brain injury through the comparison of oxiracetam for injection and piracetam for injection in clinical trials. Methods A multiple-center, randomized, double-blind,parallel study was performed on 239 patients; these patients were divided into experimental group (oxiracetam for injection, n=120) and control group (piracetam, n=119). National institutes of health stroke scale (NIHSS), Glasgow coma scale (GCS), myodynamia grading, mini-metal state examination (MMSE) were employed to evaluate the therapeutic effects; electrocardiogram and laboratory examination were performed, and the side effects were also observed. Results The scores of NIHSS,GCS and myodynamia grading after treatment in the 2 groups were all significantly higher than those before treatment (P＜0.05); however, no significant differences on these scores were noted between the experimental group and control group (P＞0.05). No serious adverse events were noted in both groups.Conclusion Oxiracetam, the same as piracetam, is safe and effective in the treatment of neurological deficits secondary to brain injury.