?Objective:To study the effects of Docetaxel on the proliferation and metastatic potential of mucoepidermoid carcinoma M 3SP 4 cells in vitro and in vivo . Methods:Inhibitory effects of Docetaxel on the proliferation and metastatic potential of mucoepidermoid carcinoma M 3SP 4 cells were investigated with cell counting,cloning assay flow cytometry, tail vein injection and submandibular gland injection of the cells into nude mice. Results:Docetaxel inhibited M 3SP 4 cells growth in a dose and time dependent way. The IC 30 and IC 50 (with 72 h exposure) of Docetaxel were 0.34 nmol/L and 0.63 nmol/L, respectively; the doubling time(h) of the cells treated with the drug at IC 30 for 7 days and of the control were 32.7 h, 43 h, respectively; the clonogenesity(%) of the control and of the cells treated with Docetaxel ( 0.05 nmol/Lor 0.1 nmol/L)were (29.2?1.4)% and (20.2?0.8)% and (2.8?0.4)%, respectively; the number of metastatic foci on lung surface in the nude mice treated with the drug at 30 mg/kg?week and in the controls were 0 and 11?3.4; the weight(g) of submandibular gland in the two groups were 0.31?0.05 and 1.20?0.23 respectively. Conclusion:Docetaxel may inhibit the proliferation and metastatic potential of mucoepidermoid carcinoma M 3SP 4 cells.

Objective To investigate the changes and clinical implications of VE-eadherin during the courseof progressive cerebral infarction.Method One hundred sixty-seven patients with acute cerebral infarction of Rong Jun General Hospital of Shandong Province and the Central People' s Hospital of Tengzhou were diagnosed in our hospital from May 2006 to July 2007,were diagnosed according to the ill%gnome criteria set by the 4th national cerebrovascular disease conference in 1995.Of them there were 102 ases with progressive cerebral infarction patients and 65 cases with non-progressive cerebral infaction.The progressive cerebral infarction patients were divided into 3 groups according to Pullicino's expressions:the big infarction focus(32 patients) ,the medium-sized infarction focus(34 patients) and the small infarction focus(36 patients) .The neurological deficits were divided into 3 groups according to the crrteria set by the 4th national cerebrovascular disease conference in 1995:light-defictits(38 patients),the moderate dificits (32 patients) and sever ditlcits (32 patients).The 65 non-progressive irffarction patients were stable without headache,vertigo and tinnitus.Arother 60 healthy subjects were entered as control group.Blood samples of all the patients' were collected at 0 h,24 h,3 d,7 d,14 d,24 d and the serum VE-eadherin by ELISA method was asaayed.All the data were analyzed by SPSS 10.0 software and One-way ANOVA was applied to intergroup comparisons for mote than two groups.Results The VF-cadherin level of patieras with progressive infarction increased in acute stage,reached the peak 3 days after onset,declined remarkably 7 days later and got nearly normalized within 21 days.The results were significantly different from those of non-progressive and controlgroup(P＜0.01).The VE-cadherin concentration was higher in patients with bigger size infarction and more sever symptoms.Conclusions The VE-cadherin level is related to the infarction size,course and the severity,and higher in the progressive group.VE-cadherin could be used for predicting prognostic of cerebral infarction and clinically valuable for treating ischemie cerebrovascular disease.

Objective: To explore the therapeutic effect of dexamethasone on proliferative retinopathy in different stages and its interaction with the vascular endothelial growth factor(VEGF).Methods: Models of oxygen-induced retinopathy(OIR) were established.Twenty C57BL/6J mice pups(7 days after birth) were randomized into 4 groups: normal control,pure OIR,early-treatment and late-treatment.Dexamethasone was administered at the dose of 0.5 mg/kg?d-1by subcutaneous injection to the early-treatment group from postnatal day 7 and to the late-treatment group from day 12,both for 5 days.Common pathological sections were made from both eyeballs of the C57BL/6J mice and detected by immunohistochemistry to count the nuclei of proliferative retinal vessel cells and investigate the expression of VEGF in the retina.Results: The nuclei of proliferative retinal vessel cells and the expressions of VEGF significantly increased in the pure OIR group as compared with the normal control(P

Objective To investigate epidemic characteristics of HCV and the distribution of naturally occurring resistance mutations of HCVNS5B in Hubei province,and provide the basis for individualized antiviral treatment for HCV infected pa-tients.Methods Collected HCV blood samples of 253 patients from March 2011 to May 2014 in Renming Hospital of Wu-han University,and determined the subtypes of HCVNS5B by first generation of sequencing technology,then compared the result with the nucleotide sequence of HCVNS5B online,located drug resistance loci and record.Results There were 4 gen-otypes and 6 different genetic subtypes in these 253 cases,namely 1a subtype 3,1b subtype 192,2a subtype 38,3a subtype 3, 3b subtypes 11 and 6a subtypes 6.Found 27 gene locus of resistance from 234 different places.The main drug resistance lo-ciare L159F (475-477)26.42%,followed by C316Y (946-948)16.98%,V321I (961-963)15.09% and S282T (844-846) 12.58%.Conclusion The epidemic characteristics of HCV in Hubei province were consistent with those of current in Chi-na;subtype 1b was the main types,followed by 2a subtype.The number of drug-resistant of HCVNS5B was more and com-plex,and presented some popular distributions.It was complex in mutations of 2a subtype,including 13 mutations of natural resistance.There were fewer mutations in subtype 1b,including sites of L159F and C316N.These results may provide the basis for antiviral therapy for HCV patients in this region.

Objective To explore the mutation spectrum of breast and ovarian cancer susceptibility gene 1/2 (BRCA1/2) which was related to breast cancer in female residents of Wuhan,and assess the relation of gene mutation and risk of suffering breast cancer.Methods 128 cases of female individuals,including 58 cases of breast cancer after surgery,70 cases of benign breast disease,and 50 femal hcalthy volunteers were selected by simple randon sampling from Department of Breast Surgery of Renmin Hospital of Wuhan University.BRCA1/2 gene was sequenced and compared with the standard template sequence to explore the possible mutations.Results In the breast cancer group,mutation emerged in 11 cases and the mutation rate was 19.0％ (11/58),including 8 cases of the BRCA1 gene mutations (3 cases of 185 del AG,5 case of 5382 ins C) and 3 cases of the BRCA2 gene mutations (2 cases of 6174 del T,1 case of C5773T) ; in the benign breast disease group,mutation emerged in 5 cases,the mutation rate was 7.1％(5/70),including 4 cases of the BRCA1 gene mutations (1 case of 185 del AG,3 cases of 5382 ins C),and 1 case of the BRCA2 gene mutation (6174 del T).There was no mutation detected in healthy control group.The mutation rate of the breast cancer group was significantly higher than that of benign breast disease group and healthy control group (x2 =4.05,10.56,P ＜ 0.05); However,there was no significant difference between benign breast disease group and healthy control group (x2 =3.73,P ＞0.05).Conclusions The mutation of BRCA1 gene (185 del AG,5382 ins C) and BRCA2 gene (6174 del T,C5773T) is in the presence of female residents in Wuhan.Furthermore,the mutation in BRCA1/2 gene increases the risk of breast cancer.

Objective To explore the accuracy and clinical application of Pyrosequencing for detection of drug resistant relevant mutation in the polymerase gene of Hepatitis B virus (HBV).Methods Compared with Sanger sequencing,the accuracy and sensitivity of Pyrosequencing were assessed.Pyrosequencing was used to determine the serum of 1164 patients with chronic Hepatitis B and its results were analyzed.Results The sensitivity of Pyrosequencing was 1 × 103 KIU/L,the same as Sanger sequencing.But Pyrosequencing was more stable and specific than Sanger sequencing to detect low rate of mutation with over 10％ mutation.The mutations couldn't be detected in 248 patients with chronic Hepatitis B,but positive results occurred in 919 patients,among them,61.5 ％ without mutation,38.5 ％ with mutation (including 47.5％ of the single locus mutation and 52.5％ of the combined mutation).Other sites had different degree of mutations except the sites of rtI169T and rtA194T; the main mutation sites in patients with chronic Hepatitis B were rtM204V/I (32.1％),rtL180M (19.8％),rtA181V/T (6.7％) and rtN236T (5.3％),in which fractional mutation had high ratio in the sites of rtA181V/T (6.7％) and rtN236T (5.3％).Conclusions Pyrosequencing has good sensitivity,specificity and accuracy and can early detect the drug resistant relevant mutation in the polymerase gene of HBV,which is suitable for monitoring patients with chronic Hepatitis B for the treatment of nucleoside analogues (acid).The different sites and frequencies of mutations may be associated with different habits of doctors for using different anti-HBV drugs.

ObjectiveTo explore the possible molecular mechanisms of β-elemene combined with cisplatin and heat therapy for killing A549 cell line.MethodsThe protein expressions of Stat3,p21 Waf1/Cip1 and Survivin were detected with Western blot after treatment with β-elemene of different concentrations combined cisplatin and heat therapy.ResultsThe protein expressions of Stat3,and pStat3 and p21Waf1/Cip1 in A549 cell line were enhanced with increasing concentrations,and there was significant difference in the expressions between high and low concentration,but Survivin protein had no change at 37°C.After adding 4 μg/mL cisplatin,the expressions of p21Waf1/ Cip1,Survivin,Stats and pStat3 were reduced at β-elemene of high concentration.At 42°C,there was no significant difference in expression of Stat3 protein at 60 μg/mL elemene,but the expressions of pStat3,p21Waf1/Cip1 and Survivin proteins had sharply declined.When using 15 μg/mL elemene combined with 4 μg/mL cisplatin,the protein expressions of Star3 and pStat3 increased,and Survivin expression decreased.ConclusionsAt temperature of 37°C,β-elemene of high concentration may inhibit growth of A549 cells by higher expression of p21Waf1/Cip1 protein,and mainly by inhibiting expressions of Stat3 and pStat3 and Survivin after combined with cisplatin.At temperature of 42°C,β- elemene of highconcentrationmaypromoteapoptosispossiblythroughinhibition ofStat3 phosphorylation and expression of Survivin protein.β-elemene of low concentration combined with cisplatin leads to synergy killing effect by reducing expression of Survivin protein.

Objective:To study the effects of Docetaxel o n the proliferation of adenoid cystic carcinoma SACC-83 cells of salivary gland. Methods:The inhibitory effects of Docetaxel on the proliferatio n of SACC-83 cells were investigated with cell counting, soft agar clonogenic a ssay, and flow cytometry. Results:With the exposure time of 24, 48 or 72 h the IC 30(nmol/L) of Docetaxel was 1.39,1.26 and 0.47, the I C 50(nmol/L) 13.02, 3.34 and 1.26 respectively; the relative antitumor acti vity (RAA) of the drug against SACC-83 cells was 330, 1 289 and 3 426 respectiv ely. After the cells had been treated for 72 h, the percentages of G 1, S and G 2 phase cells in the cell cycle in the control group were 73.8,19.8 and 6.4, in IC 30 group 65.0, 29.5 and 5.5, in IC 50 group 57.6,42.4 and 0, res pectively. The clonogenesity (%) in control, IC 30 and IC 50 groups we re 36.0?0.5,8.3?2.5 and 0.5?0.3 respectively. Conclusion:Doc etaxel may inhibit the proliferation of SACC-83 cells in a dose and time depend ent way.

Objective To evaluate the effect of 3-dimensional conformal radiotherapy (3D CRT) and prognostic factors for cervical and upper-thoracic esophageal cancer. Methods Between July 1998 and July 2001, 33 patients with cervical and upper-thoracic esophageal cancer were treated with 3D CRT(2?Gy per day, 5 sessions a week to a total dose of 66-68?Gy over 6-7 weeks). Acute toxicities and survival rates were evaluated by Kaplan-Meier method and prognostic factors were analyzed by Cox proportional hazard model. Results The 1-, 2-, 3-year local control rates were 87.9%, 75.8%, 45.5% respectively. The 1-, 2-, 3-year disease-free and overall survival rates were 72.7%, 60.6%, 30.3% and 78.8%, 66.8%, 44.2% respectively. GradeⅠ-Ⅱ acute esophagitis and bronchitis were the most common radiation side effects. Multivariate analysis revealed that the depth of primary tumor invasion, regional lymph node metastasis and tumor length were independent prognostic factors (P

Objective To evaluated the prevalence of EGFR mutations in Hubei region,to provide reliable experimental basis for reasonable screening TKI beneficiaries in clinic.Methods EGFR mutation of 253 patients diagnosed with NSCLC in Hu-bei region during 2010 to 2015 was detected by PCR-sanger sequencing and ARMS assay,to investigate the sensitivity of dif-ferent methods,to explore the frequency and clinical significance of EGFR mutation in different gender,in different histologi-cal type and different exons.Results 93 of 253 NSCLC patients harbored EGFR mutation,mainly occurred in exon 19 and 21,accounting for 53.76% and 35.38% of the total EGFR mutation rate,respectively.ADx-ARMS method showed higher sensitivity (P=0.001).The mutation detection rate of female NSCLC was significantly higher than that of male NSCLC (P=0.001).The observed incidence of EGFR mutations in patients with adenocarcinomas (38.01%)was the highest in differ-ent histological type,followed by glandular squamous cell carcinoma (30.77%),large cell carcinoma (20%)and squamous cell carcinoma (4.55%).Nonsmoking patients had a higher EGFR mutation frequency (51.6%,81/157)than those with a history of smoking (24%,12/50).Conclusion ARMS assay was more sensitive and more convenient detection method for clinical screening for EGFR-TKI treatment subpopulation.The mutation rate of female NSCLC was significantly higher than that of male NSCLC in Hubei Province.Speculated that the sexual differences in NSCLC with EGFR mutation frequencies were related to hormone levels and smoking status.