ObjectiveTo investigate the expression of stem cell marker adenosine triphosphate (ATP)-binding cassette transporter 2 (ABCG2) in the tissue and side population (SP) of a cell line A431 of human cutaneous squamous cell carcinoma(SCC).MethodsSP cells were separated by flow cytometry from cultured A431 cells.Methyl thiazolyl tetrazolium(MTT) assay was used to evaluate the proliferative ability of,reverse transcription-PCR to determine the expression of ABCG2 in,SP and non-SP cells.Immunohistochemistry (MaxVision method) was carried out to detect the expression of ABCG2 protein in tissue specimens from 10 patients with SCC.ResultsSP cells existed in cultured A431 cells,and accounted for about 1.1％ of A431 cells.The SP cells had a stronger growth and colony-forming ability than non-SP cells did.The number of cell clones formed by SP cells and non-SP cells was 114.8 ± 4.95 and 44.5 ± 3.67,respectively,per well in a 24-well plate( t =27.92,P ＜ 0.01 ).The expression level of ABCG2 mRNA was significantly higher in SP cells than in non-SP cells(t =5.22,P＜ 0.01).There existed a small number of ABCG2 positive cells in SCC tissue,and ABCG2 was mainly expressed in the cytoplasm and membrane of tumor cells.ConclusionsSP cells exist in A431 cells,which have characteristics of stem cells and highly express ABCG2.ABCG2 may be a potential stem cell surface marker of SCC.

Objective To study the antagonistic effect of N-acetylcysteine(NAC)on the damage of hepatic mitochondria of rats induced by cadmium in vitro.Methods The mitochondria were prepared from the clean Wistar rats' whole liver by using differ ential centfifugation.The mitochondria were incubated in the assay buffer containing different concentration of CdCl_2 (10,100,1 000,10 000 ?mol/L)at 37 ℃ for 1 h.The effect of NAC(500 ?mol/L)was studied at a CdCl_2 concentration of 1 000 ?mol/L.The incubation buffer was collected and the level of GSH,cytochrome C and the activity of Mn-SOD were determined. Results Compared with the control group,the level of GSH and Mn-SOD in 100,1 000,10 000 ?mol/L CdCl_2 groups were significantly decreased,the content of cytochrome C in 1 000,10 000 ?mol/L CdCl_2 groups were significantly increased(P

Objective To investigate the characteristics of pulmonary function after completed cervical spinal cord injury (CSCI) and the effect of pulmonary rehabilitation with abdominal pressure. Methods 32 male patients with CSCI were measured with pulmonary function test (PFT), and 10 of them accepted pulmonary rehabilitation with 3～5 kg pressure on their abdomen 30 min per day for 8 weeks. Results Their forced expiratory volume in one second （FEV1）, vital capacity （VC）, forced vital capacity (FVC), peak expiratory flow (PEF) were all less than 50% prediction. After 8 weeks, the pulmonary function of 10 patients who received exercise significantly improved (P<0.01). Conclusion The pulmonary function impaired in patients with CSCI. The pulmonary rehabilitation with pressure on abdomen can improve the pulmonary function.

Objective To investigate the relationship of the killer cell immunoglobulin-like receptor (KIR) gene polymorphism with Hashimoto′s thyroiditis(HT). Methods One hundred HT patients and 260 randomly matched healthy controls were enrolled to detect the KIR genotype. The genomic DNA were extracted, and 15 selected KIR genes, KIR2DL1-5, KIR3DL1-3, KIR2DS1-5, KIR3DS1 and pseudogene KIR2DP1, were determined by a polymerase chain reaction using sequence-specific primers (PCR-SSP). Results The frequency of KIR2DL5 gene was significantly lower of the patient group than that of the control group (0.200 vs 0.312, RR=0.64, P<0.01). Conclusion There may be an association between pathogenesis of HT and KIR2DL5 gene.

In order to improve the efficiency of drug screening on serotonin transporter (SERT) inhibitors, a high-throughput screening (HTS) model is established in RBL-2H3 cells. The RBL-2H3 cells are very similar to the serotonin genetic neuro, in modulation of post-receptor mechanisms and transduction pathway of SERT reactivated. Depending on a fluorescence substrate ASP+ used in detection method of inhibitor rates, it's convenient, quick, accurate and effective, not making the environmental biohazard compared with radioactive experiments. Furthermore, biological screening model combined with computer aided virtual screening technique describing high-throughput virtual screening (HTVS). Bayesian classification method and molecular fingerprint similarity were applied to virtual screening technique, for screening compounds in compound library. Some compounds have been found, and then validated further by biological screening model. Combination of HTS and HTVS improves the efficiency of screening SERT inhibitors.
Animals ; Bayes Theorem ; Cell Line ; Drug Evaluation, Preclinical ; High-Throughput Screening Assays ; Models, Biological ; Rats ; Serotonin Plasma Membrane Transport Proteins ; metabolism ; Serotonin Uptake Inhibitors ; pharmacology

Objective To summarize the clinical and pathological features of glycogen storage disease (GSD) type Ⅱ. Methods The clinical and pathological data of the 20 GSD type Ⅱ patients were reviewed. Results One patient with infantile-onset mainly presented hypotonia, muscle weakness, feeding difficulties, pulmonary infection and cardiomyopathy insufficiency and increase of serum creatine kinase (778 IU/L) and echographic evidence of hypertrophic cardiomyopathy were detected. Electromyography studies indicated a definite myopathy. Nineteen cases were late-onset, presenting a slowly progressive proximal myopathy with truncal involvement or with symptoms dominated by respiratory insufficiency. Not all muscles were equally affected. Increase of serum creatine kinase (208-2600 IU/L) was detected in 14 patients and normal level in 1 patient. Electromyography studies indicated a definite myopathy in 9 patients,with abnormal irritability in 1 patient and susceptible in 4 patients and myotonic discharge in 1 patient and no abnormalities in 2 patients. Echographic evidence of thickening of the interventricular septum and pulmonary hypertension were detected in 2 patients respectively. The common light microscopic feature of all case was a vacuolar myopathy with high glycogen content and acid phosphatase activity in the vacuoles. Conclusions GSD type Ⅱ often presents slowly progressive myopathy which often affect the toro and respiratory muscles.In most patients the serum creatine kinase level is elevated slightly. Muscle biopsy is of use to make the definite diagnosis of this disease.

Objective:To discuss the influence and curative effect of intensive lipid-lowering with atorvastatin on blood fat and ser-um inflammatory factors levels in the patients with acute cerebral infarction. Methods:Totally 94 cases of patients with acute cerebral infarction were divided into the intensive group(n=47) and the ordinary group (n=47). The patients in the two groups were given the basic medical treatment, such as reducing intracranial pressure and dehydration, controlling blood pressure and blood sugar, anti-platelet aggregation, neural protection and etc. The patients in the ordinary group were orally given 20mg atorvastatin calcium tablets, once a day, while the patients in the intensive group were additionally given 40mg atorvastatin calcium tablets, once a day, and the treatment course was 8 weeks. The changes of blood fat index and serum inflammatory factors of hs-CRP, TNF-αand IL-10 in the two groups before and after the medical treatment were detected, and the clinical curative effect was compared as well. Results:After the 8-week medical treatment, TC, TG and LDL-C levels in the two groups were declined at different degree, while HDL-C levels were in-creased at different degree (P<0. 05 or P<0. 01), and the changes in the intensive group was more notable than those in the ordinary group (P<0. 05). After the treatment, the serum hs-CRP and TNF-αlevels in the two groups were declined at different degree, while serum IL-10 levels were increased at different degree (P<0. 05 or P<0. 01), and the changes in the intensive group was more signifi-cant than those in the ordinary group (P<0. 05). Meanwhile, the total clinical efficiency in the intensive group (95. 74%) was much higher than that in the ordinary group (80. 85%, P<0. 05). Respectively 2 and 4 cases of untoward effect were appeared in the ordi-nary group and the intensive group without statistical difference between the two groups(P>0. 05). Conclusion:Intensive lipid-lower-ing with atorvastatin has significant curative effect with favorable security on acute cerebral infarction, which can obviously improve the degree of neural function defect, and the mechanism may related with reducing blood fat, serum hs-CRP and TNF-αlevels, increasing serum IL-10 levels and inhibiting topical inflammatory reactions.

Objective To research the relation between the time-dependent appearances of myofibroblasts during the repair of contused skeletal muscle in rat and wound age determination. Methods A total of 35 SD male rats were divided into the control and six injured groups according to wound age as fol-lows: 12 h, 1 d, 5 d, 7 d, 10 d and 14 d after injury. The appearances of myofibroblasts were detected by HE staining, immunohistochemistry and confocal laser scanning microscopy. Masson’s trichrome staining was utilized to examine collagen accumulation in the contused areas. Results Immunohistochemical stain-ing showed that α-SMA+ myofibroblasts were initially observed at 5 d post-injury. The average ratio of myofibroblasts was highest at 14 d post-injury, with all samples, ratios more than 50%. In the other five groups, the average of α-SMA positive ratios were less than 50%. The collagen stained areas in the contused zones, concomitant with myofibroblast appearance, were increasingly augmented along with ad-vances of posttraumatic interval. Conclusion The immunohistochemical detection of myofibroblasts can be applied to wound age determination. The myofibroblasts might be involved in collagen deposition during the repair of contused skeletal muscle in rat.

ObjectiveIn an attempt to clarify the usefulness of combined nerve and muscle biopsy in the diagnosis of neuromuscular disease when compared with traditional sural nerve biopsy.Methods Fifteen biopsies of superficial peroneal nerve (SPN) and peroneus brevis muscle ( PBM ) by one incision performed within one neurological clinic were reviewed.All patients had peripheral neuropathy while 3 of them had myopathy clinically.The diagnostic significance of SPN and PBM biopsies were classified into 3 grade: essential,helpful,no value.ResultsOf 15 SPN and PBM biopsies,7 showed essential pathological findings whichreachedthe etiologicaldiagnosis, including 5definitevasculitis, 1inflammatory demyelinating polyneuropathy and 1 amyloid neuropathy.Five biopsies are helpful for etiological diagnosis,including demyelinating neuropathy,mild inflammation,and microvascular lesion,et al.Three biopsies are of no value for etiological diagnosis which only have nonspecific change such as type 2 fiber atrophy,neurogenic atrophy and axonal degeneration et al. Finally,SPN and PBM biopsies made the definite etiological diagnosis possible in 12 patients.ConclusionsSPN and PBM biopsy improved the yield of specific pathological and etiological diagnosis of neuropathy and myopathy such as vasculitis and amyloidosis with minor trauma and side effect.Further clinical and pathological studies will be necessary for a better practice of combined nerve and muscle biopsy.

Objective: To study the influence of Centipede Acidic Protein(CAP) on the proliferation and collagen synthesis of cultured neonatal rat cardiac fibroblasts(CFb) induced by angiotensinⅡ(AngⅡ),and to explore the mechanisms of CAP on cardiac fibrosis.Methods: Neonatal rat cardiac fibroblasts were treated with AngⅡ to produce fibrosis model.The effects of CAP on proliferation of CFb were observed by MTT colorimetric assay,synthesis of collagen was observed by the hydroxyproline concentration.The NO contents were measured by Nitric acid reductase method.The c-myc expression was examined by semi-quantitative RT-PCR analysis.Results: Compared with that of control group,the proliferation,collagen synthesis and the levels of c-myc mRNA expression of CFb in the model group increased,while the NO contents decreased obviously(P