Animals ; Arteriovenous Malformations ; complications ; Child, Preschool ; Diaphragm ; blood supply ; Hemothorax ; etiology ; Humans ; Male ; Nematode Infections ; pathology ; Pleural Effusion ; etiology

Shanghai University of TCM was the first Chinese medicine university that established field-grade experimental teaching center in China.And both of the Chinese medicine and Chinese herbs experimental teaching centers became national experimental teaching demonstration centers.There is general improvement in laboratories conditions,so the experimental teaching team building is the critical factor of improving experimental teaching quality.The experimental teaching team of Shanghai University of TCM consists of excellent teachers as its backbone,and lecturers and technicians from fields in traditional Chinese medicine Chinese herbs and clinical practices.The team members cooperate with each other by setting up experimental teaching research groups to improve teaching quality,which plays an important role in building experimental teaching demonstration center.

AIM:To assess the protective role of melatonin(MEL)in a rat model of oleic-induced acute lung injury.METHODS:Twenty-four rats were randomly allocated to three groups as follows:saline(NS)injection group,oleic acid(OA)injection group and MEL plus OA injection group,the lavage protein,lung wet-to-dry weight ratio,malondialdehyde(MDA)content,superoxide dismutase(SOD)activity and lung histopathology were examined.RESULTS:(1)Injection 0.15 mL/kg of OA led to a severe acute lung injury(ALI),characterized by significantly increasing in lavage protein,lung coefficient(P＜0.01),and by histopathological alterations which presented hemorrhage,edema.thickened alveolar septum and the existence of inflammatory cells in alveolar spaces;(2)Infusion of MEL(20 mg/kg,intraperitoneally for 60 min before the oleic acid)markedly alleviated above-mentioned symptom induced by OA,consistent with decrease of MDA level(P＜0.01) and the increase of SOD activty(P＜0.01).CONCLUSION:Pre-treatment with MEL can attenuate the OA -induced ALI in rats via cleaning and preventing the formation of free radicals and further lessening the increase of alveolocapillary membrane permeability,these data suggest that MEL may be effective in the prevention of ALI.

Objective To evaluate three molecular methods for rapid identification of nontuberculous Mycobacterium(NTM).Methods Forty-one clinical NTM isolates were collected and 16S rRNA gene sequencing was used as the standard method for NTM identification.Meanwhile,the restriction fragment length polymorphism of hsp65 PCR-RFLP and hsp65 gene sequencing were used to identify NTM strains and compared with 16S rRNA gene sequencing.Results The results of 16S rRNA sequencing showed that there were nine Mycobacterium chelonae complex strains,seven Mycobacteriumfortuitum strains,seven Mycobacterium intracellulare strains,three Mycobacterium avium strains,three Mycobacterium kansasii complex strains, three Mycobacterium smegmatis strains, three Mycobacterium terrae strains, two Mycobacterium phlei strains,two Mycobacterium nonchromogenicum strains,one Mycobacterium scrofulaceum strain and one Mycobacterium arupense strain.Compared with 16S rRNA gene sequencing,hsp65 PCR-RFLP could identify nine Mycobacterium chelonae complexes and three Mycobacterium kansasii complexes to subspecies Mycobacterium abscessus and Mycobacterium kansasii,respectively; One Mycobacterium fortuitum strain and one Mycobacterium nonchromogenicum strain were different from 16S rRNA gene sequencing results ,but other isolates were the same.The coincidence was 95.1%.By hsp65 gene sequencing,only one identification of Mycobacterium hiberniae strain was different from 16S rRNA gene sequencing and the coincidence was 97.6%.And hsp65 gene sequencing could further identify nine Mycobacterium chelonae complexes and three Mycobacterium kansasii complexes to subspecies Mycobacterium abscessus and Mycobacterium kansasii,respectively.Conclusions All three molecular methods can identify NTM strains rapidly.Compared with 16S rRNA gene sequencing,hsp65 gene sequencing and hsp65 PCR-RFLP are easier to identify clinical common NTM strains(such as Mycobacterium kansasii and Mycobacterium abscessus),and can be widely used in clinical practice.

Objective To identify non-tuberculous Mycobacterium(NTM) rapidly with HAIN molecular assay genotype Mycobacterium kit,and investigate the advantages and disadvantages of this method.Methods Seventy-four clinical NTM isolates were collected from hospitals in Zhejiang and Anhui province.Clinical strains were identified with HAIN molecular assay genotype Mycobacterium kit.16S rRNA gene sequencing was used to estimate and compare with this method.Results The results of kit showed that there were thirty-one M.intracellulare strains,twelve M.chelonae strains,eight M.fortuitum strains,six M.kansasii strains,five M.avium strains,three M.smegmatis strains,two M.phlei strains,two M.scrofulaceum strains and one M.gordon strain.Four strains were identified as Mycobacterium without further identification.Eight M.tuberculosis strains were identified correctly too.Compared with 16S rRNA gene sequencing,except for four strains identified as Mycobacterium,others 70 strains got the same results as 16S rRNA gene sequencing,the coincidence was 94.59％,and it could further identify thirteen Mycobacterium chelonae complex and eight Mycobacterium kansasii complex to subspecies M.abscessus and M.kansasii,respectively.If only to identify strains under the identification range of this kit,the coincidence reach to 100％,Conclusion The method of HAIN molecular assay genotype Mycobacterium kit is simple and accurate,the time is shorter and should widely be applied clinically.

OBJECTIVE: To investigate clinical biomechanical principle of brace with stiletto needle therapy for scoliosis. METHODS: Based on design ideas of teasing needle therapy, building an experimental mechanical model was built, seven specimens with scoliosis were chosen, and treated by brace therapy and then added to stiletto needle therapy. RESULTS: The two experimental mechanical model methods could predict load of scoliosis by stiletto needle therapy, and was verified accuracy and effectiveness of model. The degree of initial scoliosis of 7 patients was (59.7±3.37)°, improved to (49.57±2.79)° by correction of brace, and (39.43±1.94)° by correction of brace with stiletto needle therapy, had significant differences(<0.05). Lateral distraction force of thoracolumbar fossa from scoliosis as V, compressive force of scoliosis as T, brace with stiletto needle therapy could save effort for 45% to 46% than that of brace, while running torque Mw and compressive torque Mv could save effort about 45% to 47%, save effort of tension torque MT of muscle and ligament for 52%, and had statistical difference(<0.05). CONCLUSIONS Experimental biomechanical model of teasing needle therapy confirmed that the therapy could significantly reduce Cobb angle, improve correction efficiency of brace and beneficial for correction effect. It is an effective treatment for scoliosis.
Biomechanical Phenomena ; Braces ; Humans ; Needles ; Pressure ; Scoliosis ; Treatment Outcome

BACKGROUNDOur previous study confirmed that oligodendrogliomas had higher frequency of chromosome 1p/19q deletion. In order to improve the diagnostic criteria and to predict the prognosis of oligodendroglioma patients, the status of chromosome 1p/19q deletion, the methylation of O(6)-methylguanine-DNA methyltransferase (MGMT), and the expression of p53 protein were evaluated and investigated in relation to patients' outcomes.

METHODSMethylation of MGMT in 73 cases was analyzed by nested methylation-specific PCR (MSP). The levels of MGMT and p53 protein were tested with immunohistochemistry. Pearson's chi-square test and Fisher's exact test were used. Multivariate and Kaplan-Meier analysis were performed to determine patients' outcomes.

RESULTSBoth oligodendrogliomas and astrocytic gliomas exhibited frequent methylation of MGMT. However, the results of MSP did not completely correspond to that of the immunohistochemical staining for MGMT. The expression of p53 protein was more frequently observed in patients without a 1p or 19q deletion in anaplastic oligodendrogliomas (P = 0.032, 0.025). In low-grade oligodendrogliomas, methylation of MGMT was more frequent in patients with 1p/19q deletion than in patients with 1p/19q intact (P = 0.038). Patients with oligodendrogliomas with 1p/19q loss of heterozygosity and p53-negative showed a longer progression-free survival.

CONCLUSIONDetection of chromosome 1p/19q status combined with p53 protein immunohistochemistry might be beneficial to improve the pathological diagnosis and to determine the prognosis of patients with oligodendrogliomas.

Adolescent ; Adult ; Aged ; Astrocytoma ; genetics ; Brain Neoplasms ; diagnosis ; genetics ; mortality ; Child ; Chromosome Deletion ; Chromosomes, Human, Pair 1 ; Chromosomes, Human, Pair 19 ; DNA Methylation ; DNA Modification Methylases ; genetics ; DNA Repair Enzymes ; genetics ; Disease-Free Survival ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Loss of Heterozygosity ; Male ; Middle Aged ; Oligodendroglioma ; diagnosis ; genetics ; mortality ; Prognosis ; Tumor Suppressor Protein p53 ; analysis ; Tumor Suppressor Proteins ; genetics

AIMTo study whether store-operated Ca2+ channel (SOC) is present in rat colonic smooth muscle cells.

METHODSIntracellular Ca2+ ([Ca2+]i) changes induced by thapsigargin- or caffeine-activated SOC channel were measured in enzymatically dissociated rat colonic smooth muscle cells with the fluorescent indicator Fura-2/AM.

RESULTSIn the absence of external Ca2+ , the sarco-endoplasmic reticulum Ca2+ pump inhibitor thapsigargin (1 micromol/L) and ryanodine receptor (RyR) activator caffeine both transiently elevated [Ca2+]i from (68.32 +/- 3.43) nmol/L to (240.85 +/- 12.65 ) nmol/L, (481.25 +/- 34.77) nmol/L. A subsequent reintroduction of Ca2+ into the extracellular solution resulted in [Ca2+]i further elevated to (457.55 +/- 19.80) nmol/L, (1005.93 +/- 54.62) nmol/L; (643.88 +/- 34.65) nmol/L, (920.16 +/- 43.25) nmol/L, respectively. And the elevated response was blocked by lanthanum (1 mmol/L), but was insensitive to L-type voltage calcium channels blocker verapamil and membrane depolarization.

CONCLUSIONSOC activated by store depletion are present in rat colonic smooth muscle cells. And we further prove the existence of such Ca2+ channels in excitable cells.

Animals ; Caffeine ; pharmacology ; Calcium ; metabolism ; Calcium Channels ; physiology ; Colon ; cytology ; Fura-2 ; metabolism ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Thapsigargin ; pharmacology

Homo sapiens longevity assurance homologue 2 (LASS2) is a novel gene isolated from a human liver cDNA library by our laboratory, and it is a human homologue of the yeast longevity assurance gene LAG1 (Saccharomyces cerevisiae longevity assurance gene). According to our previous results, LASS2 could interact with subunit c of vacuolar type H(+)-ATPase (V-ATPase), and the overexpression of LASS2 could inhibit the cell growth of a human hepatocellular carcinoma (HCC) cell line, SMMC-7721. In order to understand the role of the interaction between LASS2 and V-ATPase in HCC cell growth, we transiently transfected plasmid pCMV-HA2-LASS2 into HCCLM3, a HCC cell line without the significant expression of endogenous LASS2. The pH-sensitive fluorescence probes, BCECF and BCECF-AM, were used to measure the intracellular and extracellular H(+) concentrations of HCCLM3 cells respectively. The effect of LASS2 gene on apoptosis was evaluated with Annexin-V/FITC and propidium iodide (PI) by flow cytometry. Western blot was used to detect cytochrome c (Cyt c) in the cytosol and mitochondria, as well as pro-caspase-3 in cytosol. The results showed that the cell growth of LASS2-transfected HCCLM3 cells was significantly inhibited compared with that of the mock control. LASS2 transfection increased intracellular H(+) concentration of HCCLM3 cells, while decreased extracellular H(+) concentration. Moreover, LASS2 transfection significantly enhanced the apoptosis of HCCLM3 cells. In LASS2-transfected cells, the amounts of Cyt c increased in the cytosol, while decreased in the mitochondria. Meanwhile, the expression of pro-caspase-3 in the cytosolic extracts was decreased. These results implicate that LASS2 gene might increase intracellular H(+) of HCC cells via the interaction with V-ATPase, thereby inducing cell apoptosis through mitochondrial pathway.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Liver Neoplasms ; pathology ; Membrane Proteins ; metabolism ; RNA, Small Interfering ; Sphingosine N-Acyltransferase ; metabolism ; Transfection ; Tumor Suppressor Proteins ; metabolism ; Vacuolar Proton-Translocating ATPases ; metabolism

OBJECTIVETo study the effect of embelin on proliferation, differentiation and apoptosis of HL-60 cells and explore its possible mechanism.

METHODSDifferent concentration of embelin were used to treat HL-60 cells. Cell growth curve was analysed by MTT assay, cell apoptosis by Annexin V/PI double staining and JC-1 dye. The differentiation of HL-60 cells was evaluated by expression of CD33, CD34, CD11b and CD14. Bone marrow cells (BMC) from nine patients with acute nonlymphocytic leukemia (ANML) were also studied.

RESULTSEmbelin induced differentiation of HL-60 cells with significant increase of CD14 and CD11b expression at 33.97µmol/L for 3 days (P < 0.01). Embelin induced apoptosis of HL-60 cells in a time- and dose-dependent manner, the apoptosis rates were (9.23 ± 0.05)%, (25.86 ± 0.30)% and (39.03 ± 0.07)% respectively at 339.67 µmol/L of embelin for 12-, 24- and 48-hours treatment (P < 0.05); the apoptosis rates were (0.07 ± 0.03)%, (7.43 ± 0.30)%, (14.01 ± 0.01)%, (25.52 ± 0.03)% and (39.15 ± 0.01)% respectively at 10.19, 33.97, 101.90, 339.67 and 1019.02 µmol/L of embelin for 24-hours culture (P < 0.05). Clusters of differentiation antigen on BMC from three acute promyelocytic leukemia patients showed significant changes at 33.97 µmol/L of embelin treatment for 3 days. Embelin induced apoptosis of BMCs from all the nine ANML patients at 33.97 µmol/L for 24 hour.

CONCLUSIONEmbelin can inhibit proliferation and induce differentiation and apoptosis of HL-60 cells. The mechanism may be related to mitochondrial apoptosis pathway. Embelin at subtoxic concentration doesn't promote leukemia BMC differentiation, but at 339.67 µmol/L induces apoptosis of these cells.

Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism