Objective To investigate the immunological activity and the effect of an anti－ keratin monoclonal antibody 5G5 on the proliferation of serum－ free cultured keratinocytes. Methods The reactivity of 5G5 with cultured keratinocytes was observed by immunohistohemistry technique. The keratins extracted from the cultured keratinocytes was identified by SDS－ PAGE and their reactivities with 5G5 were assessed by immunoblot analysis. MTT was used to study the influences of 5G5 on the proliferation of cultured keratinocytes. Results 5G5 recognized the band of keratin with molecular weight of 50 000 only. Positive staining of 5G5 was found in the cytoplasm of cultured keratinocytes. The strong potential of 5G5 to inhibit the proliferation of cultured keratinocytes was shown by MTT assay with dose－ dependent manner. Conclusions The antibody which specifically recognizes the keratin with molecular weight of 50 000 may be the effective component to inhibit the proliferation of cultured keratinocytes.

Objective To study the inhibitory effects of wild-type p53 on gastric cancer cell lines BGC-7901 and to explore the importance of wild-type p53 in gene therapy for gastric cancer. Methods Adenoviral vector with exogenous wild-type p53 gene was transfected into the gastric cancer cell line BGC-7901 which harbours a mutation in codon 204 of the p53 gene.The transfer efficiency and expression effect of p53 were examined by immunohistochemistry and in situ hybridization. The growth of BGC-7901 cells in vitro was determined by cell counting and MTT assay. Flow cytometric analysis was used for observing cell cycle.Results Transfer efficiency reached 100% in BGC-7901 cells when adenovirus infection ability was over 100MOI. 3 days after transfection, the introduction of exogenous wild-type p53 into the tumor cells resulted in decreased expression of the protein of mutant p53 and increased level of mRNA of wild-type p53 gene. The growth of the BGC-7901 cells was significantly slower after transfection with wild-type p53 gene than that before transfection. The percentage of G 1/G 0 phase of the cells with exogenous wild-type p53 gene was much higher than that of the cells without exogenous wild-type p53 gene.It was 56 47% and 79 40% respectively(P

To evaluate the effects of sevoflurane and isoflurane at 1.0 minimum alveolar concentration (MAC) on intracranial pressure(ICP) in patients undergoing craniotomy.Method:Sixteen patients without pre-existing increased ICP subjected to craniotomy for pituitary adenoma or craniopharyngioma were intubated with midazolam, fentanyl and atracurium.After intubation,midazolam and atracurium were pumped intravenously for 30 min.Then,sevoflurane or isoflurane was inhaled and the patients were randomly divided into two groups:A:1.0 MAC sevoflurane:B:1.0 MAC isoflurane.In the whole procedure,PaCO_2 with an average of 5.3 kPa was maintained. ICP was measured continuously by placement of a catheter into lumbar subarachoid space.Result:Sevoflurane maintained cerebral perfusion pressure slightly better than equipotent dose of isoflurane. During sevoflurane administration at 1.0 MAC.ICP decreased slighly,then increased gradually.However,ICP remained stable in group B.Conclusion:Sevoflurane and isoflurane at 1.0 MAC,Combined with midazolam and fentanyl,can be administered safely to the patients without preexisting increased ICP.

Objective To improve the radiological diagnosis of breast cancer .Methods The authors collected 15 cases of breast carcinoma confirmed by operation and pathology. All these cases were misdiagnosed by mammography.Mammographic manifestations and data of clinic were analyzed retrospectively. Results 4 cases changed like small fibroadenoma, 3 cases showed microcalcification, 3 cases showed localized mammary gland architecture distrotion,2 cases showed multiple small patchy shadows in the hyperplasic mammary gland, 2 case showed small focus of increased density, 1 case no abnormal sign, 4 cases among of them can not be palpated mass .Conclusion The combination between palpation and radiology is valuable for diagnosis of breast carcinoma and decreasing misdiagnosis.

Objective To assess the clinical efficacy ofMaxing-Shigan decoction in patients with severe pneumonia due to phlegm-heat obstructing the lung.Methods Sixty patients with severe pneumonia in the TCM Department of Dalian Municipal Central Hospital from January to December 2013 were randomly divided into a control group and a treatment group by random number table method, 30 in each group. The control group was treated with anti-infection, fluid infusion and mechanical ventilation. On the basis of treatment in the control group, the treatment group was also givenMaxing-Shigan decoction through nasogastric gavage. The mean arterial pressure (MAP), lactic acid (Lac) and PaO2/FiO2 were compared before and after the treatment in both groups. The scores of the Clinical Pulmonary Infection Score (CPIS) and the Acute Physiology and Chronic Health Evaluation II (APACHEⅡ) were compared between the two groups.Results The total effective rate in the treatment group was significantly higher than that in the control group (80.0%vs.73.3%;χ2=4.800,P=0.029). In both groups, MAP (treatment group: 102.4 ± 11.9 mmHgvs. 70.4 ± 6.4 mmHg, t=12.972,P=0.000; control group: 101.2 ± 12.5 mmHgvs. 69.8 ± 6.6 mmHg,t=12.1672,P=0.000), PaO2/FiO2 (treatment group: 265.4 ± 19.7 mmHgvs. 78.6 ± 10.5 mmHg,t=45.833,P=0.000; control group: 124.8 ± 13.4 mmHg vs. 79.1 ± 11.1 mmHg;t=14.385,P=0.000) after the treatment were significantly higher than before the treatment, and the Lac level were significantly lower (treatment group: 1.04 ± 0.42 mmol/Lvs. 6.37 ± 2.27 mmol/L,t=12.65, P=0.000; control group: 3.88 ± 1.45 mmol/Lvs. 6.39 ± 2.32 mmol/L,t=5.025,P=0.000). The improvements in Lac (t=10.304,P=0.000) and PaO2/FiO2 (t=32.323,P=0.000) in the treatment group were superior to the control group. The scores of the CPIS (treatment group: 2.2 ± 1.5vs. 6.7 ± 1.7,t=10.872,P=0.000; control group: 4.5 ± 2.7 vs. 6.8 ± 1.8,t=3.882,P=0.000) and APACHEⅡ(treatment group: 5.3 ± 2.1vs. 13.8 ± 3.8,t=10.723,P=0.000; control group: 9.3±3.5vs. 13.7 ± 3.5,t=4.869,P=0.000) after the treatment were significantly lower than before the treatment in both groups, and the changes of CPIS (t=4.079,P=0.000) and APACHEⅡ(t=5.368,P=0.000) in the treatment group were greater than those in the control group.ConclusionsMaxing-Shigan decoction has definite therapeutic effects for severe pneumonia due to phlegm-heat obstructing the lung..

Objective To investigate the intervention of Chinese herb Shengjiang San(SJS)for the imbal-ances of serum Th1/Th2 cells and related regulatory factors in sepsis patients. Methods Fifty-five sepsis patients were randomly divided into two groups:conventional treatment group of 27 cases and SJS group of 28 cases. Nine cases of healthy volunteers were enrolled as control group in the study. Cases of conventional group were treated with western medicine only and cases of SJS group were treated with both western medicine and Chinese herb SJS (100 mL twice one day). The therapy course of both groups was 3 days. Score of Chinese medical syndromes ,se-rum level of leukocyte count,C reactive protein(CRP)and procalcitonin(PCT),serum T-bet,GATA-3,Th1&Th2 cells in the proportion of whole CD4+Th cells and Th1/Th2 ratio were compared respectively in each group be-fore and after treatment. Results There were significant differences between SJS group and conventional group in score of Chinese medical syndromes,serum level of leukocyte count,T-bet,GATA-3,Th1&Th2 cells in the pro-portion of whole CD4+Th cells and Th1/Th2 ratio(P<0.05 or P<0.01). There were no significant differences be-tween the two groups in serum level of CRP,PCT and GATA3. Compared with control group,there were signifi-cant differences in all indicators except GATA3 of the two groups(P<0.01). Conclusion Chinese herb Shengji-ang San has an effective benefits to Chinese medical syndromes,inflammatory reaction,the imbalance of serum Th1/T2 and related regulatory factors(T-bet&GATA-3)in sepsis patients.

Establishment of combination of disease and syndrome animal model is an ideal technology platform in TCM research. Currently, many scholars have prepared matching IBS animal models combined typical animal models with etiology, which em-bodies the advantages of syndrome differentiation and promotes the development of combination disease and syndrome animal model. Even though, there still exists some problems about sub-jective evaluation on syndromes, paying more attention on dis-ease and less on syndromes. Therefore, we should deepen the pathogenesis of diseases and TCM syndromes research on animal model replication and improve the method of construction of combination of disease and syndrome animal model through the biological research of combination of disease and syndrome sys-tem. By this way, the idea of preparation animal models will be more mature and the scientific connotation of TCM will be pres-ented more exactly.

Objective To investigate therapeutic effect of the Banxia-Baizhu-Tianma decoction combined with simvastatin for phlegm and blood stasis obstructing hyperlipidemia. Methods A total of 120 patients with phlegm and blood stasis obstructing hyperlipidemia from Dalian central hospital from June 2010 to December 2013 were divided into 2 groups according to random digits table:60 patients in a control group treated with simvastatin 20 mg/d,and 60 patients in a treatment group treated with Banxia-Baizhu-Tianma decoction on the basis of the control group. After 6 weeks treatments, the clinical efficacy, serum levels of total cholesterol(TC), triglyceride(TG), high density lipoprotein cholesterol(HDL-C), low density lipoprotein cholesterol(LDL-C), high sensitive C reactive protein(hs-CRP),high mobility group protein B1(HMGB1), alanine aminotransferase(ALT), γ-glutamyl transpeptidase(γ-GT), interleukin 8(IL-8)and superoxide dismutase(SOD) were evaluated in both groups. Results The total effective rate was 91.7%(55/60) and 73.3%(44/60) in the treatment group and the control group, respectively, with significant difference (χ2=5.772, P<0.05). After treatment, the serum levels of TC, TG, HDL-C, hs-CRP, HMGB1 in both groups were significantly lower than those before the treatment (in the treatment group t value were 12.355, 5.984, 32.825, 13.806, 14.661 and in the control group t value were 7.553, 2.177, 3.064, 10.281, 9.019, P<0.05 or P<0.01), LDL-C concentration significantly increased (t in the treatment group=3.382, P<0.01;t in the control group =2.388, P<0.05). After the treatment, the serum levels of TC, TG, HDL-C, hs-CRP, and HMGB1 in the treatment group were lower than those in the control group (t value were 7.173, 3.418, 2.377, 3.967, 8.352, P<0.05 or P<0.01);there was no significant difference in LDL-C concentration between the two groups (t=1.630, P=0.106);the levels of ALT, GT, IL-8 and SOD in the treatment group showed significant difference compared with those before the the treatment (t value were 19.053, 9.592, 12.909, 10.377, P<0.01), while in the control group, exceptγ-GT, other parameters showed significant difference after the treatment compared with before the treatment (t value were 2.253, 5.876, 3.266, P<0.05). Compared with the control group, these parameters in the treatment group were significantly different after the treatment(t value were 27.872, 10.244, 6.954, 6.145, P<0.01). Conclusions Banxia-Baizhu-Tianma decoction combined with simvastatin has good lipid-lowering effect. It may significantly lower inflammatory factor levels, reduce inflammation, and enhance antioxidant capacity.

OBJECTIVETo investigate the contribution of mitochondrial pathway in the apoptosis induced by puerarin (PUE) in pulmonary artery smooth muscle cells.

METHODCultured rat pulmonary artery smooth muscle cells (PASMC) were intervened by high, middle and low dose of puerarin (1.5 x 10(-3), 1.5 x 10(-4), 1.5 x 10(-5) mol x L(-1)). The change of mitochondrial membrane potential was observed. Western blot detected the expression of apoptosis-related gene Caspase-9, Bax and Bcl-2 protein.

RESULTCompared with the control group, mitochondrial membrane potential significantly decreased in puerarin groups. Puerarin can enhance the expression of Caspase-9 and Bax protein, decrease the expression of Bcl-2 protein. Puerarin also has a concentration-dependent on the induction of PASMC.

CONCLUSIONPuerarin can induce PASMC apoptosis through mitochondrial pathway.

Animals ; Apoptosis ; drug effects ; Caspase 9 ; metabolism ; Cells, Cultured ; Isoflavones ; pharmacology ; Male ; Mitochondria ; drug effects ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; physiology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Pulmonary Artery ; cytology ; drug effects ; Rats ; Rats, Wistar

Objective To study the effect of a Chinese medicine, Huanglian Jiedu Decoction ( HLJDD) , on ath-erosclerotic plaque, inflammatory factors and regulatory T cells in ApoE-/- mice. Methods High fat diet was given to ApoE-/- mice to establish an atherosclerosis model. 40 male ApoE-/- mice were randomly divided into five groups:model group, simvastatin group, low, moderate and high dose HLJDD groups (n =8). HLJDD was intragastrically administered in a dose of 3. 5, 7. 0, or 14. 0 g/( kg?bw) once dailg for 16 weeks. The dose of simvastatin was 5. 0 g/( kg?bw). An-other 8 male C57BL/6J mice were taken as control group. At the end of the 29-week experiment, all of the mice were sacri-ficed. The aortic plaques, level of blood lipids, inflammatory factors, Tregs number and the level of Foxp3 mRNA were de-tected and analyzed by ELISA, flow cytometry and RT-PCR, respectively. Results Compared with the control group, the aortic plaques were much larger in the model group, and the levels of TC, TG, LDL-C, IL-6, hs-CRP, and TNF-α were significantly higher than those in the control group (P<0. 01 for all). Meanwhile, the levels of HDL-C, IL-10, TGF-βand Foxp3 mRNA were much lower than those in the control group ( P<0. 01 for all) , and the Tregs numbers were less than that in the control group (P<0. 01). HLJDD regulated the blood lipids in ApoE-/ - mice and decreased the levels of IL-6, hs-CRP, andTNF-α, however increased the levels of IL-10, TGF-β and Foxp3 mRNA. At the same time, it in-creased Tregs number in the ApoE-/ - mice. Compared with the model group, the difference was statistically significancet (P<0. 01). Conclusions HLJDD can significantly alleviate the aortic plaque damages in ApoE-/ - mice. It may be re-lated to the up-regulation of Tregs, which can lead to decrease the expression of serum pro-inflammatory factors such like IL-10, hs-CRP and TNF-α.