BACKGROUND: Both antioxidant and cytokine can induce the differentiation of umbilical cord blood-derived mesenchymal stem calls (UCB-MSCs) towards neuron-like cells in vitro. It remains unclear regarding how to select an inductor that has the ability of either neuro-protection like cytokines or powerful antioxidation. After wide screening, baicalin has been included in this study. OBJECTIVE: To observe the effects of baicalin on in vitro purification, amplification, and differentiation towards neuron-like cells of hUCB-MSCs.DESIGN, TIME AND SETTING: A randomized, controlled, cytological in vitro observation was performed at the laboratory of Department of Neurology, First Affiliated Hospital of Nanchang University between February and April 2005. MATERIALS: Ten portions of UCB were collected from healthy full-term normal delivery pregnant women aged 23-25 years old. Baicalin with purity > 95% was purchased from Department of Pharmaceutics, Xiangya Medical College of Central South University. Antioxidant additive J -mercaptoethanol was provided by Sino-American Biotechnology Company, China. METHODS: The collected UCB was anticoagulated with heparin to separate mononuclear cells. After concentration adjustment (1 ×109/L), UCB mononuclear cells were purified and amplified with dulbecco's modified eagle's medium containing fetal bovine serum (0.2 volume fraction), glutamine, B27, granulocyte colony-stimulating factors, and stem cell factors. According to antioxidant additive application, 4 groups were set: baicalin, blank control, β -mercaptoethanol, and baicaUn+ β -mercaptoethanol. Cells in each group were cultured for a total of 4 consecutive weeks. MAIN OUTCOME MEASURES: (1) Detection of CD 34 and CD 29 immunoreactive expression on days 7, 14, 21, and 28 after cryopreservation. (2) Cellular morphology observation. (3) Detection of surface antigen expression of MSCs by flow cytometry. (4) Detection of neuron-specific enolase (NSE), microtubule associated protein 2 (MAP-2), and glial fibrillary acidic protein (GFAP) expression after 4 weeks of culture by immunocytochemistry. RESULTS: O Compared with prior to cryopraservation, trypan blue exclusion rate of UCB-MSCs was significantly reduced on days 7, 14, 21, and 28 after cryopreservation (P < 0.05). (2) Morphological observation: UCB mononuclear cells adhered to the wall 2-3 days after culture, reached a peak level at 2 weeks, and formed a confluence of approximately 80%-90% 3 weeks after culture; at this time, all UCB-MSCs displayed a spindle shaped appearance. Four weeks later, in the baicalin group, some spindle-shaped UCB-MSCs began to present shrinkage, with slender processes on the cell edge, and some UCB-MSCs tended to be spherical-, conical-, and triangle-shaped appearance, with many slender processes on the pseudopodia. In the β-mercaptoethanol and baicalin+β -mercaptoethanol groups, an increasing number of cells defoliated and died with culture time in addition to above-mentioned appearances. (3) Four weeks after culture, cells were positive for CD45 in the blank control group, while cells in the remaining groups were positive for CD29 and CD 83, in particular in the baicalin+ β -mercaptoethanol group, followed by the baicalin group, and lastly the β -mercaptoethanol group. Significant difference in CD29 and CD 83 immunoreactivity exhibited between groups (P < 0.01). No CD34 immunoraactive calls were found in each group. (4) Four weeks after culture, NSE and MAP-2 immunoreactive expression was significantly lower in the blank control and β -mercaptoethanol groups than in the baicalin group (P < 0.01). The percentage of cells expressing GFAP was lower than 1% in each group. CONCLUSION: 100 μmol/L baicalin can promote the in vitro amplification of UCB-MSCs in a time-dependent manner and also can induce the differentiation of UCB-MSCs towards neuron-like cells in vitro to some extent.

Objective To study the factors related to hyperuricaemia in type 2 diabetic patients and to investigate the effect of serum uric acid levels on the metabolic syndrome.Methods During March 2007 and March 2008 731 cases were divided into 2 groups according to their uric acid levels,then the relationship between serum uric acid and metabolic parameters and the complications in two groups were observed.Results There was a positive correlation between uric acid levels and triglycerides,body mass index,blood pressure(P

BACKGROUND: Present studies have verified that Baicalin has protective effects on various brain damage in the nervous system.OBJECTIVE: To study the possibility of intravenous transplantation of human umbilical blood mesenchymal stem cells (hUBMSCs) and Baicalin after hypoxic-ischemic brain damage (HIBD) in neonatal rats.DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Laboratory of the Department of Neurology, First Affiliated Hospital, Nanchang University from February 2007 to January 2008.MATERIALS: Totally 10 umbilical blood samples from healthy full-term pregnant women were obtained from the Department of Obstetrics, First Affiliated Hospital, Nanchang University. A total of 85 clean Sprague Dawley neonatal rats aged 7 days were randomly assigned to a normal control group (n=15), a model group (n =20), a cell transplantation group (n =25), a cell transplantation + Baicalin group (n =25).METHODS: The umbilical blood mononuclear cells were isolated by the gelatin sedimentation + density gradient centrifugation method, and amplified in vitro. Cells at the fifth passage were used for transplantation. Cells were labeled by DAPI at 6-12 hours before use. Neonatal rats in the model, cell transplantation and cell transplantation + Baicalin groups were used to establish HIBD models. Rats in the blank control group were left intact. At 2, 3,4, 5 weeks following model induction, rats in the cell transplantation and cell transplantation + Baicalin groups were injected with DAPI-labeled hUBMSCs (5-10 μL/g) via caudal vein at the density of 1 ×10~9/L. From the first day of transplantation, rats in the cell transplantation + Baicalin group were injected with 120 mg/kg Baicalin via intraperitoneal injection, once a day, for three successive days.MAIN OUTCOME MEASURES: The following parameters were measured: brain tissue lesion, DAPI-positive cell number, location of hUBMSCs following transplantation.RESULTS: Lesion rate of brain tissue was significantly lower in the cell transplantation + Baicalin group compared with the model and cell transplantation groups at 4 weeks following transplantation (P < 0.05). Compared with the cell transplantation group,DAPI-positive cell number was significantly increased in the cell transplantation + Baicalin group at 1, 2, 4 weeks (P < 0.01). From the 3~(rd) week following model induction, abundant DAPI-labeled cells were found surrounding the lesion site, without obvious boundary integrated with the host brain. Few DAPI-positive hUBMSCs were found in non-ischemic region. At 4 and 5 weeks following model induction, DAPI-positive cells were significantly decreased in the lesion site.CONCLUSION: The third week following HIBD is an optimal time for cell transplantation. Baicalin can make a large number of hUBMSCs across the blood-brain barrier to distribute and scatter around the disease focus integrated with host brain tissue.

Based on the results of oral glucose tolerance test( OGTT )and the levels of 1-h plasma glucose ( 1 hPG),793 subjects were classified into three groups:583 with NGTN ( normal 1 hPG in OGTT),127 with NGT1 H( higher 1 hPG in OGTT) and 83 with IGT( impaired glucose tolerance).NGT1H group had large waist circumference,higher body mass index,fasting plasma glucose( FPG),triglyceride,and lower high density lipoprotein-cholesterol than those of NGTN group.NGT1 H group had higher homeostasis model assessment insulin index ( 1.2 ± 0.6),lower homeostasis model assessment β3 ( HOMA-β ) (4.5 ± 0.7 ) and insulinogenic index (2.1 ±0.7) than those of NGTN group(0.5 ±0.6,4.8 ±0.7,2.7 ±0.9,respectively,all P ＜0.05 ).HOMA-β of NGT1 H group was higher than that of IGT group(4.5 ±0.7 vs.4.4 ±0.6,P ＜0.05 ).The results indicate that 1 hPG in OGTT may identify a condition of glucose metabolic abnormalities characterized by insulin resistance and reduced β-cell function.

Objective To investigate the role of N-methyl-D-aspartate receptor one(NMDA-R1 )in visceral hypersensitivity forming of Sprague-Dawley rat model induced by transient Trichinella spiralis intestinal infection.Methods Forty male Sprague-Dawley rats were divided into healthy control group,visceral hypersensitivity group,0.9％ NaCl group and MK-801 group.The rats in visceral hypersensitivity group,0.9％NaCl group and MK-801 group were given Trichinella spiralis to establish infection model.After eight weeks,the rats in 0.9％NaCl group and MK-801 group were given 0.9 ％ NaCl and MK-801 respectively.The abdominal muscular electrical activity of each group was observed under 20,40,60 and 80 mm Hg (1 mm Hg=0.133 kPa) colorectal distension pressure.After abdominal muscular electrical activity recorded,rats were sacrificed and the colon specimens were collected.The expression of NMDA-R1 at protein level was detected by Western blot.The data were analyzed by LSD test and Bivariate correlation.Results There was significant correlation between areas under curve (AUC) of abdominal muscular electrical activity electromyography and distension pressure in healthy control group,visceral hypersensitivity group,0.9％ NaCl group and MK-801 group (r=0.823,0.618,0.913,0.889 respectively,all P＜0.01).Under 20,40,60,80 mm Hg distension pressure,there was significant difference in AUC between MK-801 group and visceral hypersensitivity group (LSD test,all P＜0.05).The AUC of MK-801 group was lower than that of healthy control group,but there was significant difference (LSD test,P =0.029) only when distension pressure was at 80 mm Hg.The integral optical density value of NMDA-R1 protein of MK-801 group (1.238 ±0.210) was significantly lower than that of visceral hypersensitivity group (2.231±0.450) and 0.9％NaCl group (2.104±0.220) (LSD teat,P=0.025,0.046).The JA value of NMDA-R1 protein of visceral hypersensitivity group and 0.9％NaCl group was significantly higher than that of healthy control group (LSD test,P =0.007,0.014).Conclusion The visceral hypersensitivity forming was correlated with the high expression of NMDA-R1,and it could be adjusted by inhibiting NMDA-R1 expression in enteric nervous system.

Objective To investigate the role of synaptic plasticity on the formation of visceral hypersensitivity induced by transient intestinal infection in rats. Methods Thirty male Sprague-Dawley rats were divided into normal control, acute infection and chronic infection groups with 10 each. The area under curve (AUC) of electromyography (EMG) in 10 s was used to evaluate the visceral sensitivity induced by different eolorectal distention (20,40,60 and 80 mmHg). Histological change of the colon was evaluated by H-E staining. Synaptic uhrastrueture such as synaptic cleft and synaptic vesicles was observed using transmission electron mieroseope. The mRNA and protein expressions of synaptophysin and postsynaptic density protein-95 (PSD-95) were examined by RT-PCR and Western blot, respectively. significantly higher than those of normal controls(P=0. 012, 0. 005, respectively ). In contrast, AUC of acute infection were significantly lower than those of normal controls ( P = 0. 018,0. 012, respectively ). Under the distention of 20 and 80 mmHg, no significant difference was observed among three groups (P= rats compared to normal controls(23.45±4.10 vs. 9.10±2.42, P=0. 027),but there was no statistical difference between chronic infection rats and normal controls (13. 95±7.96 vs. 9.15±2.42, P=0.78). increased. In acute infection rats, mitochondria cristae disappeared, synaptic vesicles and the length of controls, mRNA and protein of synaptophysin in ileocecum, proximal colon and distal colon were significantly increased in chronic infection rats (P＜0. 05 ), but decreased in acute infection rats with no significant difference. Compared with controls, no significant downregulation was noted in the expression protein expressions of PSD-95 were both increased in chronic infection rats (P＜0.05), and decreased in acute infection rats (P＜0.05). Conclusion Synaptic plasticity plays an important role in the formation of visceral hypersensitivity induced by transient intestinal infection in rats.

Objective To assess the antimicrobial resistance and resistance-related mutations in rpoB and katG genes in Mycobacteria tuberculosis isolates from patients with cutaneous tuberculosis. Methods Seven strains of Mycobacteria were isolated from lesions or secretions of patients with cutaneous tuberculosis and identified as M. tuberculosis. Proportion method was used to test the susceptibility of M. tuberculosis isolates to rifampicin, isoniazid, streptomycin and ethambutol. PCR and sequencing were performed to analyze the mutations in rpoB and katG genes. Results Of the 7 isolates of M. tuberculosis, 1 was resistant to rifampicin,isoniazid and ethambutol simultaneously, and the other 6 were sensitive to rifampicin, isoniazid, streptomycin and ethambutol. All the 7 isolates were positive for the amplification of rpoB and katG genes by PCR. DNA sequencing revealed two mutations at codon 531 (TCG to TTG) and codon 315 (AGC to ACC) in the multi-drug resistant strain, which were absent in the other 6 strains. Conclusion Multi-drug resistance has emerged in M. tuberculosis isolates from patients with cutaneous tuberculosis, which is likely to be related to improper treatment.

Objective To detect the expression of Th17 pathway-related genes in patients with syphilis serofast reaction and to investigate the mechanism of Th17 cells in syphilis serofast reaction. Meth-ods Peripheral blood samples were collected from patients with syphilis serofast reaction ( n=8 ) , patients who were syphilis-seronegative after treatment (n=8) and healthy subjects (n=8). Total RNA was extrac-ted from each blood sample and then reversely transcribed into cDNA. PCR-Array analysis was performed to quantify the expression levels of Th17 pathway-related genes. Results The expression levels of genes with a fold change >2 (up or down regulated) were defined as differentially expressed. (1) Compared with the control group, the patients with syphilis serofast reaction showed increased expression of genes encoding fork-head box protein 3 (Foxp3) and IL-10, but decreased expression of genes encoding C-C motif chemokine 22 (CCL22), colony stimulating factor 2 (CSF2), CSF3, chemokine (C-X-C motif) ligand 6 (CXCL6), IL-17A, IL-17D, IL-21, IL-23R, IL-9, interferon regulatory factor 4 (IRF4), RAR-related orphan receptorα( RORα) , RAR-related orphan receptor γ ( RORγ) and signal transducer and activator of transcription 3 (STAT3). (2) Compared with the seronegative syphilis group, the expression levels of genes encoding Foxp3 and IL-10 in patients with syphilis serofast reaction were up-regulated, while the expression of genes encoding CCL22, CSF2, CSF3, IL-17A, IL-21, IL-23R, IRF4, RORγ and STAT3 were down-regulated. (3) The expression levels of genes encoding CXCL6 and IL-9 in seronegative syphilis group were lower than those in control group. Conclusion The abnormal expression of Th17 pathway-related genes might relate to the pathogenesis of serofast state of syphilis.

Objective To explore the role of synaptic plasticity on the formation of visceral hyper- sensitivity induced by acute restraint stress in rats.Methods Twenty male Sprague-Dawley rats were randomly divided into control group and acute restraint stress group(model group).Visceral hypersensi- tivity was made by acute restraint stress for 1 h.The colorectal distension(CRD) with different pressure were performed and the abdominal electromyography(EMG) was recorded.The visceral sensitivity was determined by the frequency of EMG.The ultrastrueture of synapse was observed with transmission electron microscope.The expression of synaptophysin was measured by RT-PCR and Western-blot. Results①The frequency of EMG was significantly correlated with CRD pressure(control group,r=0.992, P=0.008;model group,r=0.978,P=0.022).The frequencies of EMG in model group(at 40,60 and 80 mm Hg) were significantly more than that in control group(P value=0.043,0.024,0.038,respectively).②There were more synaptic vesicles accumulated in presynaptical terminal.The post synaptic density was increased in model group compared to control group.③In the proximal and distant colon,the expressions of rnRNA and protein of synaptophysin were higher in model group (P

OBJECTIVETo evaluate the effect of hemoperfusion on tetramine poisoned patients.

METHODSThree tretramine poisoned cases treated with hemoperfusion were selected. The samples during and after hemoperfusion were collected and analyzed by gas chromatography.

RESULTSTetramine concentration at the inlet of the artificial kidney kept the same level during hemoperfusion. After hemoperfusion, the tetramine concentration in patient plasma changed little in 72 hours. 1.03-1.55 mg of tetramine was adsorbed by the instrument of hemoperfusion after two hours' hemoperfusion.

CONCLUSIONAlthough hemoperfusion was not so effective to reduce blood tetramine concentration in patients, it could clear about 1 mg tetramine for one time.