Objective:To analyze the expression of E-cadherin in the epithelial-mesenchymal transition (EMT) induced by transforming growth factor-β1 (TGF-β1) in neuroblastoma.Methods:TGF-β1(1 μg/L, 5 μg/L, 10 μg/L), was applied to SK-N-SH cells in vitro compared with the blank control group.EMT-related genes mRNA and protein expression were detected by carrying out real-time PCR assays and Western blot.A scratch test and migration assay were performed to verify the alteration of SK-N-SH cell migration capacity.Data collected from 18 cases of neuroblastoma patients were selected from the Department of Hematology Oncology, Shanghai Children′s Hospital from January 2008 to December 2012.The expression of E-cadherin in the tumor tissue of the neuroblastoma patients after operation was detected by immunohistochemistry.The clinical features and survival prognosis of these patients were analyzed. Results:Compared with the control group, after SK-N-SH cells were treated with TGF-β1(1 μg/L, 5 μg/L, 10 μg/L), real-time PCR assays and Western blot revealed that the mRNA(0.603±0.081, 0.606±0.008, 0.716±0.166 vs.1.000) and protein expression levels(0.855±0.026, 0.600±0.017, 0.495±0.011 vs.1.000) of E-cadherin were significantly decreased ( F=8.144, P=0.040; F=74.810, P<0.001), while the mRNA(2.132±0.167, 3.494±0.017, 4.184±0.021 vs.1.000) and protein expression levels (1.175±0.053, 1.189±0.058, 1.225±0.106 vs.1.000)of α - smooth muscle actin were significantly increased ( F=547.300, P<0.001; F=68.810, P=0.007), suggesting that EMT changes occur in cells.Scratch test and Transwell migration assay revealed that the number of migrating cells increased obvious with the treatment of TGF-β1 (5 μg/L) ( t=16.070, P=0.040). The 10-year overall survival(OS) rates of neuroblastoma patients with E-cadherin strong positive expression, positive expression, weak positive expression and negative expression in the pathology were (77.78±13.86)%, (75.00±21.66)%, (25.00±21.65)% and 0, respectively ( F=8.160, P=0.040). Conclusions:TGF-β1 can induce the EMT in SK-N-SH cells and increase cell migration.The decrease expression of E-cadherin in neuroblastoma patients is closely associated with clinical progress and recurrence or metastasis of the disease.

Objective To detect the changes of visceral sensitivity in rats presenting intestinal dysbacteriosis and the expressions of tight junction protein (ZO-1)and Toll-like receptor 4 (TLR4)so as to explore the effect of intestinal dysbacteriosis on visceral sensitivity and the possible mechanisms.Methods We randomly divided 30 male SD rats of SPF grade into normal control group (n = 12 )and dysbacteriosis group (n = 18 ).Rats in dysbacteriosis group were administered with lincomycin hydrochloride (300 mg/mL),1 mL each time per rat once a day for 7 consecutive days;those in normal control group were fed with the same amount of saline.On the eighth day,six rats were randomly selected from normal control group and dysbacteriosis group respectively to detect whether the model was successful.After the model was successfully constructed,the remaining 12 dysbacteriosis rats were randomly divided into the negative control group and the probiotics intervention group with 6 in each.Rats in the intervention group were given probiotic bifidobacterium triple viable capsules (Bifico)orally,one capsule with 1/3 mL of saline,1 mL each time per rat once a day for 7 consecutive days;those in the negative control group received the same amount of saline.On the eighth day,fresh feces was cultured for flora to detect visceral sensitivity by abdominal withdrawal reflex (AWR),the mRNA and protein expressions of ZO-1 and TLR4 in the colon,and the expression of serum inflammatory cytokines IL-10 and TNFα.Results The expression of ZO-1 in the colon was significantly lower in the rats of dysbacteriosis group than those in the control group,and the expression of TLR4 was also significantly increased.Correspondingly,the expression of pro-inflammatory factor TNFα in the serum of the rats in dysbacteriosis group was significantly increased,while that of anti-inflammatory factor IL-10 was significantly lower than in the control group (P <0.05).Furthermore,compared with dysbacteriosis group,the expression of ZO-1 was increased significantly and TLR4 was decreased in probiotics group in varying degrees. Similarly,the expression of TNFα was obviously lower while that of IL-10 in the serum was higher (P < 0.05 ). Conclusion Inhibiting the expression of ZO-1 and increasing the expression of TLR4,thus leading to chronic low-grade inflammation, may be one mechanism of visceral hypersensitivity caused by intestinal dysbacteriosis. Probiotics may restore the dysbacteriosis and thus improve visceral hypersensitivity.

Objective To find out the detection level of the blood routine test in clinical laboratories of basic medical institutions in Hefei ,and to analyze the results of inter‐laboratory comparison among township health centers and community health service cen‐ters in Hefei and explore the main factors .Methods Forty‐three township health centers and community health service centers were randomly selected to conduct field investigations and take blood routine test inter‐laboratory comparison .Results Both 41 .9% of the passing rate and the average score 72 .37 points in Inter‐laboratory comparisons were significantly lower than the An‐hui province clinical inspection center(94 .1% and 95 .97 points) ,the differences were statistically significant(P<0 .05);Comparing to the results of the Anhui province clinical inspection center ,there was statistically significant difference on parameter(WBC ,RBC , Hb ,HCT ,PLT) average pass rate of blood routine test(P<0 .05);the personnel primary education was low ,18 .80% of the staff in clinical laboratories were not professionals ;most of blood analyzers were domestic and 53 .49% of all instruments had been used for more than 5 years;the overall laboratory quality management level was low .Conclusion The blood routine test detection level in clinical laboratories of basic medical institutions in Hefei was far below than that of secondary and tertiary medical institutions .The daily laboratory internal quality control should be strengthened and the quality management system should be improved gradually .

Objective To compare the infectivity between laboratory adapted human inununodefi- ciency virus(HIV-1) and primary HIV-1 isolates for different mucosal epithelial cell lines. Methods Mu-cosal epithelial cells Caco-2, T-84, HeLa and lymphocyte MT-4 were infected with laboratory adapted HIV-1 SF33 and 2 primary HIV-1 isolates (02010561, 02010141). Culture supernatant and cells were collected respectively on 3-4 days interval after virus inoculation. The former was tested for HIV-1 antigen P24 level and viral load, and the latter was tested for total viral DNA and integrated viral DNA. Results All 3 virus strains could infect MT-4 cells and integrate into their genome. Only HIV-1 SF33 could infect Caco-2 cells but could not integrate into their genomic DNA. Both HIV-1 SF33 and 02010561 infected HeLa cells but only integration of HIV-1 SF33 was detected. All the 3 HIV-1 strains infected T-84 cells but only the integra-tion of HIV-1 SF33 and 02010141 was observed. Conclusion Although laboratory adapted and primary HIV-1 strains are able to infect human mucosal epithelial cell lines, transient or productive infection estab-lished in different mucosal epithelial cells is dependent on the character of cells and virus strains.

The purity of 4,4′-dimethoxy-5,6,5′,6′-bis (methylenedioxy)-2′-morpholine methylenebiphenyl-2-methyl formate methanesulfonate (IMH), a new drug for fatty liver treatment, was determined through differential scanning calorimetry (DSC). Analysis of two-factor non repeatability method was performed in the investigation the effects of two factors (heating rate and sample weight) on purity determination. The DSC experimental parameters were optimized as follows: heating rate was 10 ℃·min^-1, temperature range was 150-300 ℃, sample weight was 2.0-4.1 mg, and N₂ flow rate was 80 mL·min^-1. The linear correlation coefficient (r) of this DSC method was 0.999 8. The RSD value (n = 6) of precision was 0.03%. The standard value and uncertainty of the purity results of the multiple batches of IMH drugs were (99.74 ± 0.29)%, (99.91 ± 0.28)%, (99.90 ± 0.28)%, and (99.81 ± 0.28)% with inclusion factor (K) of 2 and confidence probability (P) of 0.95. The results were basically consistent with the results of the mass balance method. The DSC mehod is a simple, rapid and accurate method, and provides a new reference method for determining the purity of IMH drugs, improves the accuracy and reliability of purity determination.

OBJECTIVETo observe the effect of Bushen Huoxue Granule (BHG) on dopamine (DA) neurotransmitter and dopamine transporter (DAT) in the brain of patients with Parkinson's disease (PD) as an adjunctive therapy.

METHODSNinety-four PD patients were randomly assigned to two groups, 47 in each group. Madopar was given to all as the basic treatment group. The placebo was given to those in the control group while BHG was given to those in the treatment. The therapeutic course for all was three months. Before and after treatment DA levels in the brain of patients were detected by encephalofluctuograph (EFG) technique. Changes of DAT in the striatum of patients in the treatment group were detected by positron emission tomography (PET) and region of interest (ROI) analysis.

RESULTS(1) Before treatment the DA level was lower in the two groups than the normal value, showing significant difference (P < 0.01), but with no significant difference between the two groups (P > 0.05). After treatment the DA level obviously increased in the two groups, showing significant difference from that before treatment (P < 0.01). No significant difference existed in the DA level in the two groups when compared with the normal value (P > 0.05), but with significant difference between the two groups (P < 0.05). Better results were obtained in the treatment group than in the control group. (2) The DAT radioactive accumulation inside the striatum increased obviously in the treatment group after treatment. ROI analysis showed the total ratio of striatum/cerebellum before and after treatment was 1.86 +/- 0.32 and 2.61 +/- 0.53 respectively, showing statistical difference (P < 0.05).

CONCLUSIONBHG could improve the DA level of PD patients, and increasing DAT contents in the striatum, thus playing a role in effectively treating PD.

Aged ; Brain ; metabolism ; Dopamine ; metabolism ; Dopamine Plasma Membrane Transport Proteins ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; drug therapy ; metabolism ; Phytotherapy

Acute Disease ; Adult ; Chromatography, Gas ; Female ; Humans ; Lung ; chemistry ; Phosphines ; analysis ; blood ; poisoning

OBJECTIVETo provide a new material for producing the Rhodiolasachalinensis products, the effect of methyl jasmonate (MeJA) on callus biomass and effective compound accumulation of Rhodiolasachalinensis was studied.

METHODThe calluses-cultured in 3 L-air lift balloon type bioreactor were treated with MeJA after 20 d of bioreactor culture and the effect of MeJA concentration and treatment days on callus biomass, salidroside or polysaccharide accumulation and superoxide dismutase (SOD) and peroxidase (POD) activities were investigated.

RESULTThe callus biomass was not significantly different after MeJA treatment (125) for 0-6 d but obviously decreased after 6 d treatment. The maximum salidroside or polysaccharide contents and SOD or POD activities were found after 4 d treatment of MeJA. MeJA concentration significantly affected callus biomass and effective compound accumulation, biomass decreased at MeJA concentrations higher than 125 μmol x L(-1). However, the effective compound contents were determined at higher MeJA concentration, and the highest salidroside and polysaccharide accumulation was found at 225 and 275 μmol x L(-1) MeJA, respectively and the maximum SOD and POD activities was found at 225 μmol x L(-1) MeJA. The effective compound contents in callus were compared with field-grown plants. Salidroside contents in calluses were 1.1-fold and 2. 4-fold more than in plant roots and stem or leave, respectively. Polysaccharide content in calluses were 3. 6-fold and 8.0-fold more than in plant roots and stem or leave, respectively.

CONCLUSIONSalidorside and polysaccharide in Rhodiolasachalinensiscalluses improved by MeJA treatment, 225 μmol x L(-1) MeJA and 4 d treatment were optimal. The effective compound contents in callus were obviously higher than in field-grown plants. Therefore, bioreactor culture is efficient for obtaining mass effective compounds of Rhodiolasachalinensis by culturing calluses. This method could provide an alternative material source for production of Rhodiolasachalinensis products.

Acetates ; pharmacology ; Biomass ; Bioreactors ; Cyclopentanes ; pharmacology ; Glucosides ; metabolism ; Oxylipins ; pharmacology ; Peroxidase ; metabolism ; Phenols ; metabolism ; Polysaccharides ; metabolism ; Rhodiola ; metabolism ; Superoxide Dismutase ; metabolism

Objective To compare the mRNA level of macrophage inflammatory protein-3α(MIP-3α) in 3 different mucosal epithelial cell lines. Methods RNA standards were prepared by in vitro transcription. One-step real-time RT-PCR was established and optimized using TaqMan EZ RT-PCR Core Reagents with TaqMan probes and primers specific to human MIP-3α mRNA sequence. The specificity of one-step real-time RT-PCR method was confirmed by sequencing the PCR products. The sensitivity and reproducibility of the method was examined by repeating the test 8 times with the same sample. Results The one-step real-time RT-PCR with a wide detection range is sensitive, reproducible. It was found that MIP-3α mRNA level in Caco-2 and T-84 cells was much higher than that in the HeLa cells. Conclusion High level of MIP-3α mRNA could be found in mucosal epithelial cells and difference in transcription level of MIP-3α may exist in epithelial cells from different mucosa.

SHIV-CN97001 played an important role in assessing the immune effect and strategy of the AIDS vaccine which included genes of the predominant prevalent HIV-1 strain in China. In this study, SHIV-CN97001 was in vivo passaged serially to construct pathogenic SHIV-CN97001/rhesus macaques model. To identify variation in the gp120 region of SHIV-CN97001 during passage, the fragments of gp120 gene were amplified by RT-PCR from the plasma of SHIV-CN97001 infected animals at the peak viral load time point and the gene distances (divergence, diversity) were calculated using DISTANCE. The analysis revealed that the genetic distances of SHIV-CN97001 in the third passage animals were the highest during in vivo passage. It had a relationship between viral divergence from the founder strain and viral replication ability. The nucleic acid sequence of the V3 region was highly conservative. All of the SHIV-CN97001 strains had V3 loop central motif (GPGQ) and were predicted to be using CCR5 co-receptor on the basis of the critical amino acids within V3 loop. These results show that there was no significant increase in the genetic distance during serial passage, and SHIV-CN97001 gp120 gene evolved toward ancestral states upon transmission to a new host. This could partly explain why there was no pathogenic viral strain obtained during in vivo passage.