The combination of TCM and WM has developed for more than 50 years,with fruitful achievements from drug study to talents training.However,it also encountered several problems on the way of development.In the course of the practice of the combination of TCM and WM,we find that for the real combination of TCM and WM,we must base on the fully understanding the theories,respective lacks and the aim of the combination of TCM and WM,moreover,the combination is not to make TCM become WM,but the combination on the level of theory.

ObjectiveTo study the expression of Twist in invasive ductal carcinoma of breast and its relationship with cell proliferation and angiogenesis.MethodsThe expression of Twist and Ki-67 was detected in 60 cases with invasive ductal carcinoma of breast by immunohistochemistry.Ki-67 index and microvascular density(MVD) were calculated.ResultsThe positive expression rate of Twist was 56.7％ (34/60) in invasive ductal carcinoma of breast.Ki-67 index and MVD in the patients with positive expression of Twist was higher than those in the patients with negative expression of Twist[(57.05 ± 16.37)％ vs.(25.32 ± 16.16)％,(34.30 ± 12.25)％ vs.(23.04 ± 10.45)％,P＜ 0.05 ].ConclusionsThe overexpression of Twist is found in invasive ductal carcinoma of breast.To stimulate cell proliferation and angiogenesis may be one of the pathways of Twist to contribute to the invasion and metastasis of breast cancer.

Objective To investigate the effects and mechanisms of all-trans retinoic acid(ATRA)on the proliferation and cell cycle of lung carcinoma cell line A549.Methods The A549 cells were treated with ATRA at the dosages of 5,10,50 ?mol/L for 1-7 d.The proliferation of A549 was assessed by MTT method and cell cycle was analyzed by flow cytometry.The expressions of CDK4,Rb and p-ERK1/2 were assessed by Western blotting.CyclinD1 mRNA was analyzed by SYBR-PCR amplification.Results ATRA obviously inhibited the proliferation of A549 cells,and the cell cycle was arrested in G0/G1 phase.The expression of p-ERK1/2 protein and CyclinD1 mRNA on A549 cells were decreased.Conclusion ATRA might inhibit the proliferation of A549 cells through down-regulating p-ERK1/2 protein and CyclinD1 mRNA.

Objective To observe the role of preoperative lung function test in predicting the risk of postoperation pulmonary complications in patients with chronic obstructive pulmonary disease (COPD) accepting non chest operations. Methods 80 patients accepting non-invasive chest operations during Oct 2006 to May 2013 in the third affiliated hospital of SYSU were studied retrospectively. All the patients accepted lung function test 1 week before operation. Based on the lung function records, patients were divided into 2 groups. 40 of them in COPD group, 40 in control group. The incidence rate of postoperation pulmonary complications in different group and the relationship between the severity of lung function decreasing and the rate of postoperation pulmonary complications were investigated. The differences of the American Society of Anesthesiologists (ASA) Physical Status Classification, body mass index, smoking index, length of stay, hospitalization costs between the 2 groups were also studied. Results The incidence rate of postoperation pulmonary disease in COPD group was 30% (12/40) while the rate in control group was 12.5% (5/40), the statistic difference was significant (P = 0.046). There was remarkable relationship between the severity of lung function decreasing and the rate of postoperation pulmonary complications(P=0.005), patients with mild to moderate lung function decreasing would be safer in operation, but patients with severe lung function decreasing would be in high risk(r=-0.451). Patients in COPD group were older than the control group, but there were no significant difference on body mass index, smoking index, length of stay, hospitalization costs between the 2 groups (P > 0.05). There was no relationship between ASA physical status classification and postoperation pulmonary complications. Conclusion Incidence of postoperation pulmonary complications in patients with COPD is high, which mainly manifests as pneumonia. It was important to test the lung function before non-invasive chest operations, especially in patients with COPD(P>0.05).

Objective To understand the direct medical expense for surgical patients with splenomegalic advanced schisto?somiasis and its influencing factors,so as to provide evidences for relevant departments to improve the rescue strategy of ad?vanced schistosomiasis. Methods The data about the expenses of patients with splenomegalic advanced schistosomiasis hospi?talized in Xiangyue Hospital affiliated to Hunan Institute of Schistosomiasis Control from January 2010 to August 2014 were col?lected,the hospitalization expense and hospital stays of the patients were analyzed,and the factors influencing the hospital ex?penses were analyzed by the univariate and multi?factor analyses. Results From January 2010 to August 2014,totally 249 cas?es were hospitalized in the hospital,their average hospital stays and hospital expenses were 28.92 d and 18 896.13 Yuan,and both of them were increased year by year. Among all the kinds of expenses,the constitution ratios of the medicine expenses were the highest,and those in the 5 years were all above 44%. The results of the univariate and multi?factor analyses showed that the hospital stays,the amount of intraoperative bleeding,liver function classification,postoperative complications,age,portal hy?pertensive gastropathy were the influencing factors of the hospital expenses. Conclusion Presently,the burden of the direct hospital expenses of the patients with splenomegalic advanced schistosomiasis is still heavy. The government should further im?prove the proportion of the compensation of medical assistance and perfect the medical aid scheme. Meanwhile ,the hospitals should strengthen the management and standardize medical behavior to reduce the hospitalization expenses of the patients.

Objective To establish an isolation method for solid GTC in peripheral blood using EpCAM antibody-linked nanobeads and evaluate the sensitivity of the method and its application significance. Methods Five, ten, twenty, fifty and one hundred MCF7 (breast cancer), KYSE70 (esophageal cancer), BxPC-3 (pancreatic cancer) and 9811P (stomach cancer) cells were added into 7. 5 ml erythrocyte lysed peripheral blood obtained from healthy volunteers respectively. EpCAM antibodylinked nanobeads were used to enrich cancer cells. The recovery rates of the in vitro added cancer cells were evaluated by fluorescence microscopy. Then, the untreated thirty cases of esophageal cancer (six cases at stage Ⅰ and Ⅱ, twenty-four cases at stage Ⅲ and Ⅳ), thirty-five cases of breast cancer (fifteen cases at stage Ⅰ and Ⅱ , twenty cases at stage Ⅲ and Ⅳ), thirty cases of pancreatic cancer (five cases at stage Ⅰ and Ⅱ , twenty-five cases at stage Ⅲ and Ⅳ), thirty-three gastric cancer (thirteen cases for stage Ⅰ and Ⅱ ,twenty cases at stage Ⅲ and Ⅳ) were enrolled to enrich the peripheral blood CTC. Thirty healthy volunteers and thirty gastritis patients served as two groups of control. Meanwhile the enriched CTC was identified by IF and HE staining. FISH was used to analyze the copy number of chromosome 8 and chromosome 20 in two hundred esophageal cancer, breast cancer, pancreatic caner and gastric cancer CTC. Results After DAPI staining and mixing with 7.5 ml peripheral blood from healthy donors, the average cell recovery rates of KYSE70, MCF7, BxPC-3 and 9811P cells evaluated under fluorescence microscope were 87%, 87%, 86% and 88% (within group), and the recovery rates of 5 gradient dilution levels were 88%, 85%, 87%, 88% and 87% (intergroup). With a high sensitivity, this method was able to isolate one cancer cell in 107 white blood cells of peripheral blood. The positive rates of more than 2 CTC in the peripheral blood detected by this method were 50% (15/30) of esophageal cancer, 63% (22/35) of breast cancer, 70% (21/30) of pancreatic cancer and 61% (20/33) gastric cancer patients respectively,but no CTC was detected in the peripheral blood of healthy volunteers and gastritis patients (P = 0. 000).The aneusomy of chromosome 8 and chromosome 20 were found in 80% esophageal cancer, 75% breast cancer, 65% pancreatic cancer and 59% gastric cancer. Conclusions The CTC isolation technique with EpCAM antibody-linked nanobeads is sensitive and accurate. The aneusomy of chromosome 8 and 20 is frequent in CTC from esophageal cancer, breast cancer, pancreatic cancer and gastric cancer.

AIM:Panton-Valentine leukocidin(PVL)is a pore-forming toxin secreted by Staphylococcus aureus epidemiologieally associated with the often-lethal necrotizing pneumonia.Until now,the mechanisms of pathogene-sis of PVL leading to the fatal pulmonia remains undefined and also acquired plenty of the toxins is difficult.In the present study，we obtain recombinant staphylococcal F and S components of the Panton-Valentine leukocidin by gene engineering and evaluate its biological activity in vitro,which provides an experimental basis for the further studies of its biological func-tion and its toxicity in pneumonia.METHODS:The full-length of F and S components of PVL gene amplified from the strain of Staphylococcus aureus DNA by hiSh-fidelity PCR was cloned into prokaryotic expression vector pET22b(+),and the vector was transformed into BL21(DE3)plysS to construct a prokaryotie expression system.The integrity of the opening-reading frame of each construct was verified by DNA sequencing.The recombinant PVL(rPVL)was induced by1.0 mmol/L IPTG.The expressed products were identified by SDS-PAGE and the fusion proteins(6His-LukS-PV and 6His-LukF-PV)were purified from lysates of transfeeted E.coli cells by affinity chromatography on nitrilotriacetic acid columns.The eytolytie activity was tested by incubation of rPVL with human polymorphonuclear neutrophils(PMNs)in vitro.RESULTS:The nueleotide sequence of the cloned PVL gene was the same as that of reported in GenBank.E coli BL21(DE3)plysS containing recombinant vectors grow at 37℃causes some proteins to accumulate as inclusion bodies.while incubation at 30℃led to a significant amount of soluble active proteins which accounted for about 31.7％ of the total bacterial protein.The relative molecular weight showed on SDS-PAGE profile was consistent with the expected value which the LukS-PV protein was about 34 kD.and the LukF-PV protein was about 35 kD.The purified rPVL was obtained and its cytolytic activity to PMNs was demonstrated.CONCLUSION:The genes of 1ukS-PV and lukF-PV are successfully cloned into plasmid pET22b(+)and expressed in E coli respectively.which provide a basis for analyzing the toxicity related to the diseases and further studies about the pathogenesis of PVL.

Objective To identify differentially expression of microRNAs associated with expression of estrogen receptor α(ERα)and progesterone receptor(PR)between type Ⅰ and type Ⅱ endometrial adenocarcinoma.Methods Two kinds of endometrial adenocarcinoma cell lines,Ishikawa and KLE,was transplanted into node mice and biopsied to identify the expression of ERα,PR and p53,and test their response to estrogen and progesterone.Cultured the two cell lines under the estrogen-free and progesteronefree circumstance,total RNA was isolated to identify the differentially expressed microRNAs by microarray for prediction the microRNAs which target ESR1 and PGR by software miRANDA and TargetScan,and then was validated by real-time PCR in two cell lines cultured both in vivo and in vitro and ten specimens from patients.Results Ishikawa cell line was confirmed from type Ⅰ endometrial adenocarcinoma.KLE cell line was confirmed from typeⅡendometrial adenocarcinoma.One hundred and twenty-six differentially expressed microRNAs between the two cell lines were identified by mieroRNA microarray,among of which may target ESR1 inchded hsa-miR-100,99a,and may tgrget PGR included hsa-miR-378,768-3p-The differential expression of hsa-miR-100,99a,378,768-3p identified by microarray between Ishikawa and KLE in vivo and in vitro was equal to that by real-time PCR,while Hsa-miR-100 was significantly down expressed in type Ⅰ group specimens compared to type Ⅱ group(P＜0.01).Conclusion Hsa-miR-100 is significantly down-expressed in type Ⅰ endometrial adenocarcinoma compared to type Ⅱ,which may be a great potential to target ESR1.

Objective To study influencing factor of mechanical ventilation (MV) time in patients withchronic obstructive pulmonary disease (COPD). Method Totally 128 patients with COPD and respiratory failurerequiting ventilation support were retrespectively investigated. The clinical characteristics of patients before and af-ter intuhation during the respiratory intensive care unit (RICU) stay were recorded and analyzed, t- test, X2 testand stepwise logistic regression analysis were used for statistical analysis. Results In the 128 patients, 61%,20%, and 9% of the patients required MV longer than 7, 14 and 21 days, respectively. There were no significantdifferences in the history of COPD, smoking, lung function, and complications between patients with MV>7 d, 14d, 21 d and patients with MV<7 d, 14 d, 21 d. Multiple regression analysis showed APACHE Ⅱ score (OR:2.3; 95% Ca: 1.2-5.7, P = 0.02) was an independent factor for MV > 7 days, while shock (OR: 0.7;95% CI: 1.0-1.9, P = 0.04) and decreased serum albumin (OR: 0.4, 95% CI: 0.2-0.8, P = 0.003)were an independent factors for MV > 21 days. The ventilator-associated pneumonia (VAP) was the most impor-rant risk factor for the time of MV. Conclusions APACHE Ⅱ score, serum albumin levels, hock and VAP arethe main factors affecting MV time in patients with COPD.