AIM:To investigate the effect of pretreatment of stem cell factor(SCF) and granulocyte colony-stimulating factor(G-CSF) on the proliferation and the differentiation of mesenchymal stem cells(MSCs) into cardiomyogenic cells.METHODS:The MSCs,isolated primarily from bone marrow,and purified by passage culture,were obtained from the adult rats of four groups:the rats were pretreated by 5 daily injections of SCF;the rats were pretreated with G-CSF;the rats were pretreated with SCF and G-CSF;the rats were treated without any intervention.The 4th passage of MSCs was labeled by DAPI and cellular cycle analysis was conducted by flow cytometry before co-culture.The neonatal rat cardiomyocytes cultured for 3 days were co-cultured with DAPI-MSCs.The percentage of the differentiation of MSCs into cardiomyogenic cells during the five co-culture days was analyzed.The morphologic changes of MSCs and the proteins expression of cardiac myosin heavy chain(MHC) and troponin T(TnT) were recorded respectively with digital microscope camera system and immunofluorescence technique.The percentage of the differentiation of MSCs into cardiomyogenic cells was also calculated.RESULTS:The percentage of MSCs in G0/G1 phase in SCF/G-CSF group was significantly lower than that in SCF group,G-CSF group and the control group.The percentage of MHC protein-positive MSCs in SCF/G-CSF group was markedly higher than that in SCF group,G-CSF group and the control group,and that in SCF group and G-CSF group was significantly higher than control group.The percentage of TnT protein-positive MSCs in SCF/G-CSF group,SCF group and G-CSF group was significantly higher than that in control group.CONCLUSION:SCF and G-CSF show the ability to stimulate the proliferation of MSCs and induce MSCs to differentiate into cardiomyocytes.The combination of using SCF and G-CSF is more effective than using only SCF or G-CSF.

Arrhythmias, Cardiac ; physiopathology ; Child ; Electrocardiography ; Humans ; Male ; Tachycardia, Ventricular ; physiopathology

Objective To investigate the changes of blood coagulation indices during the percutaneous cor-onary intervention(PCI), so as to provide evidence for anticoagulation therapy. Methods 34 acute coronary syn-drome patients were enrolled,whose blood sample was taken at baseline,1 h,4 h,24 h,and 48 h after PCI operation. Thrombin-antithrombin complex(TAT), vWF, protein C, antithrombin, fibrinogen, D-dimer were tested. The result was processed by statistical software. Results After PCI, TAT continued to rise, protein C and antithrombin drop down,the vWF and D-dimer rose with TAT,and fibrinogen transiently drop down. Conclusion The thrombin pro-duces more quickly after PCI,its time phrase is coordinated with vWF and D-dimer,at the same time the anticoagula-tion system drops down. High anticoagulation at least maintains within 48 h and then tends to be normal.

[ ABSTRACT] AIM: To study the protective effects of cannabinoid CB2 receptor agonist JWH133 on rat acute lung injury induced by paraquat (PQ).METHODS: Male Sprague-Dawley rats (n=72) were randomly divided into 4 groups.PQ group:PQ was administered intraperitoneally at the dose of 20 mg/kg;Low-dose JWH133 pretreatment group ( L-JWH133 group):JWH133 (5 mg/kg, ip) was administered 1 h before PQ exposure;high-dose JWH133 pretreatment group ( H-JWH133 group):JWH133 (20 mg/kg, ip) was administered 1 h before PQ exposure;control group:1 mL sa-line was administered intraperitoneally.Arterial blood, bronchoalveolar lavage fluid (BALF) and lung tissue samples were collected at 8 h, 1 d and 3 d after PQ exposure.PaO2 and the levels of TNF-αand IL-1βin BALF were measured via blood gas analyzer and ELISA, respectively.The pathological changes and lung injury scores were assessed at 3 d after PQ expo-sure.NF-κB and AP-1 protein levels were also determined by Western blotting.RESULTS:The decrease in PaO2 , struc-tural injury of the lung tissues, interstitial pulmonary edema, and the increase in IL-1βand TNF-αin BALF were observed in PQ-treated rats compared with control group.JWH133 pretreatment reduced the degree of lung tissue injury, decreased the levels of IL-1βand TNF-αin BALF and the NF-κB and AP-1 protein expression in the lung tissue compared with PQ group, especially in H-JWH133 group.CONCLUSION:CB2 receptor agonist JWH133 inhibits NF-κB and AP-1 protein expression in the lung tissues, and reduces the secretion of IL-1βand TNF-αin BALF after paraquat exposure, thus atten-uating paraquat-induced acute lung injury.

Objective To find out the physical state of the human papillomavirus ( HPV) genome in hepatoma cell line HepG2 cells and the regulation of HPV late capsid protein 1 ( L1) expression and to explore the nature of the cytoryctes in HepG2 cells.Methods E2 and E6 in HPV18 were detected by PCR to evaluate the physical state of HPV18 genome .HepG2 L1 expression was detected by ELISA , light microscropy and electron microscrope immu-nohistochemistry assays , Western blot assay using HPV L 1 mice monoclonal antibody .L1 mRNA in HepG2 cells was detected by reverse transcriptional PCR ( RT-PCR) .Results PCR assay displayed that HPV DNA was inte-grated with HepG2 genome.ELISA assay showed that HPV L1 was present in lysate of HepG2 cells.Light micros-cropy demonstrated strong positive reaction in HepG2 cells.In microscopy, in the cytoplasm of partial HepG2 cells, there were lumpish cytorrhyctes materials which consists of very small and uniform particles and these parti -cles were marked by HPV L1 antibody labeled by colloidal gold .Western blot analysis showed a band at 56 ku dis-trict and it was L1 specific strap which demonstrated HPV 18 L1 was present in HepG2 cells.RT-PCR assay demon-strated the presence of L1 mRNA in HepG2 cells.Conclusions HepG2 cells are HPV18-positive HPV DNA ge-nome is integrated with HepG2 cells.HepG2 cells can express L1.The cytorrhyctes in HepG2 cells are composed of HPV18 L1 indicating that L1 can be expressed in HepG2.

Objective To investigate the toxicity of nasal membreane and ciliary of the Magnolia biondii Pamp volotile oil nanometer bangosome.Methods Toad palate and rat nasal membrane were used as experimental material,physiological saline and hydrochloride ephedrine as negative control.The Magnolia biondii Pamp volotile oil nanometer bangosome on ciliary movement were carried out using in vitro and electron microscope technique.Results The Magnolia biondii Pamp volotile oil nanometer bangosome had little cilitoxicity to toad palate and rat nasal membrane.Conc(?)sion The Magnolia biondii Pamp volotile oil nanometer bangosome had little cilitoxicity to membrane.

BACKGROUND: Mesenchymal stem cells (MSCs) can differentiate into myocardial cells in vitro and in vivo, but the amount is small and directed differentiation rate is low.OBJECTIVE: To explore the effect of stem cell factor (SCF) on promotion of MSCs differentiating into myocardial cells.DESIGN: Opening experiment.SETTING: Institute of Cardiovascular Disease, Xianning College and Department of Cardiology, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was performed at the Experimental Center of Institute of Cardiovascular Disease, Xianning College from October 2003to August 2004. A total of 20 infant SD rats aged 1-2 days were selected for culture of myocardial cells. Another 25 clean adult SD rats were selected and randomly divided into SCF group, blank control group with 12 rats in each group. The left one rat was used for MSCs extraction, culture and purification.METHODS: ①MSCs were labeled with 4', 6-diamidino-2-phenylindole,dihydrochloride (DAPI) before co-culture. SCF was given into rats of the SCF group by subcutaneous injection successively for 5 days, 20 μg/kg per day. Isolated MSCs were co-cultured with myocardial cells that had been cultured for 3 days. Saline of the same volume was given in the blank control group by subcutaneous injection. MSCs without any intervention were co-cultured with myocardial cells that had been cultured for 3 days.②Expressions of MHC α/β and troponin T were recorded and measured with digital micro-camera shot and immunofluorescence technique, respectively. Percentage of DAPI labeled MSCs differentiating into myocardial cells was measured.MAIN OUTCOME MEASURES: ①Growth of MSCs, ②detection of labeling rate of DAPI on MSCs, ③analysis of activity and purity of co-cultured myocardial cells, and ④differentiation of MSCs into myocardial cells after co-culture.RESULTS: ①Bone marrow cell suspension was inoculated in plastic petri dish. Round cells scattered at the bottom of bottle. At hour 24 a few long fusiform shape adhered cells appeared following liquor change, and at day 4 many fusiform shape adhered cells appeared, which entered logarithm increased period. At days 8-12 80% were confluence. The proliferation of passage cells was more rapid. With liquor change and passage, the MSCs were purified gradually. ②Labeling rate of DAPI on MSCs was 100%. ③At 3-day in vitro culture of myocardial cells the percentage of beat cells was 63%, with beat frequency of 40-60 times per minute. Percentage of positive cells of troponin T expression was 75% examined with immunocytochemical technique. ④At co-cultured days 2 and 3, positive percentage of DAPI labeled MSCs expressing MHC α/β was significantly higher in the SCF group than in the blank control group (P ＜ 0.01). At the co-cultured days 3, 4 and 5, positive percentage of DAPI labeled MSCs expressing troponin T was obviously higher in the SCF group than in the blank control group (P ＜ 0.01).CONCLUSION: SCF has markedly accelerating effect on proliferation and differentiation of MSCs.

Objective To evaluate the safety and efficacy of a domestic recombinant human tumor necrosis factor receptor type Ⅱ- IgG Fc fusion protein (rhTNFR-Fc)for the treatment of moderate to severe psoriasis vulgaris. Methods A multicenter, randomized, double blind, parallel-group, positive drug-controlled clinical trial was conducted. According to random numbers generated by a hierarchical segmentation method using the SAS 9.2 software, patients with moderate to severe psoriasis vulgaris were randomly divided into two groups to be injected with two kinds of domestic rhTNFR-Fc under the trade names of Anbainuo(test group)and Yisaipu(control group)respectively at a dose of 25 mg twice a week for 12 consecutive weeks. The primary endpoint was the proportion of patients achieving a 50%, 75% and 90% reduction in psoriasis area and severity index(PASI50, PASI75 and PASI90)at week 2, 6 and 12 after initiation of the treatment. Adverse reactions were also recorded. Statistical analysis was carried out by using the chi-square test, Fisher′s exact test, two-sample t-test, and noninferiority trials with the software SAS 9.2. Results A total of 180 patients were enrolled in this study from 5 centers, and 174 completed this trial, of whom, 88 were assigned to the test group and 86 to the control group. Analysis of the full analysis set (FAS)revealed no significant differences in PASI50(75.6%(68/90)vs. 82.2%(74/90), P > 0.05)or PASI75(51.1%(46/90)vs. 50.0%(45/90), P > 0.05) between the test group and control group, but a significant increase in PASI90 in the test group compared with the control group (30.0% (27/90)vs.16.7% (15/90), χ2 = 4.472, P < 0.05)at week 12. Drug-related adverse reactions included elevation of transaminases, leukopenia, upper respiratory infections, injection-site reactions, abnormalities in urine routine test and purified protein derivative (PPD)skin test results, etc. There was no significant difference between the two groups in the incidence rate of adverse reactions(χ2 = 0.188, P > 0.05), most of which were mild, and subsided spontaneously or after appropriate treatment. Conclusion The domestic rhTNFR-Fc (trade name:Anbainuo)25 mg twice a week for 12 weeks is effective and safe for the treatment of moderate to severe psoriasis——————————vulgaris.

Objective To explore the effect of surgical treatment on primary gastrointestinal non-Hodgking lymphoma(NHL) in children.Methods Nine cases of clinical and follow-up data of primary gastrointestinal NHL were studied retrospectively to evaluate the effect of surgical treatment on primary gastrointestinal NHL in children.Results Seven cases were male and 2 cases were female.The mean age was(5.59?3.27)years old.The clinical manifestation included abdominal mass (7 cases),abdominal pain (5 cases),fever (2 cases),haematemesis and melena (2 cases),constipation (1 case) and paroxysmal abdominal pain with vomiting (1 case).Nine cases were diagnosed as primary gastrointestinal NHL,including 1 case of intussusception,1 case of acute appendicitis,2 cases of gastrointestinal obstruction,2 cases of gastrointestinal bleeding and 3 cases of abdominal mass.One case received the operation of intussusception reduction,tumor resection and intestinal anastomosis.One case received appendectomy.One case received the operation of tumor biopsy and transverse colon colostomy.Six cases received laparotomy.Six cases were diagnosed as Burkitt lymphoma.One case was anaplastic large cell lymphoma and 2 cases were diffuse large B-cell lymphoma.One case was at stage Ⅰ,1 case was at stage ⅠE,2 cases were at stage Ⅱ,3 cases were at stage ⅡE and 2 cases were at stage Ⅲ.Nine patients had received operation.One case died after operation and 8 cases had received combined chemotherapy.The 1 and 3 years survival rates were 75.0% and 37.5%,respectively.Conclusions Acute abdomen is often the first symptom of primary gastrointestinal NHL in children and comprehensive surgical treatment is an effective procedure for it.

Objective To investigate the acute toxicity, subacute toxicity, nasal membrane local toxicity of the Magnolia biondii Pamp volatile oil nanometer bangosome. Methods Kunming mice and SD rats were selected as experimental material. Kunming mice were used in the acute and subacute toxicity tests by intragastric administration of Magnolia biondii Pamp volatile oil nanometer bangosome with different dosage and different time, and SD rats were used in the nasal membrane local toxicity test by nose dropping, while the control group was treated with intragastric administration of or nose dropping with normal saline for the same dosage. The treatment course lasted fifteen days. At the end of the tests, the general condition, routine blood test, function index of live and kidney, organ humid weight index, histological changes of liver and kidney and ultra microstructure change of rat nasal membrane were obtained and compared with the control group. Results In the acute toxicity test, the daily maximum tolerant dosage by intragastric administration was equal to 222.7 times of the clinical routine, with no marked toxic reaction. In the subacute toxicity test, the general condition, blood test, organ humid weight index and histological changes of live and kidney in different dosage groups were similar to the control group. While in the function index of live and kidney, uric acid was stepped down in the middle and large dose groups, and total bilirubin was decreased in the large dose group. The nasal membrane local toxicity test revealed that there was little change in the ultra microstructure of rat nasal membrane.Conclusion The Magnolia biondii Pamp volatile oil nanometer bangosome may have little toxicity by intragastric administration and nose dropping.