Angioplasty, Balloon, Coronary ; Chronic Disease ; Coronary Disease ; therapy ; Coronary Occlusion ; therapy ; Humans

BACKGROUND:Several studies have demonstrated that the number of alveolar epithelial cells decreases gradually as pulmonary fibrosis develops,but the reasons remain unknown.The expression of P16,CyclinD1,and CDK4 might be abnormal and P16-CyclinD1/CDK4 call cycle regulatory pathway may play an important role in the progressive reduction of alveolar epithelial cells during pulmonary fibrosis.OBJECTIVE:To investigate the dynamic expression of P16,CyclinD1,and CDK4 in the pulmonary tissue of bleomycin-inducad pulmonary fibrosis mice.METHODS:Pulmonary fibrosis was induced in 6-week-old KM mice by intratracheal injection of bleomycin(BLM group).The control mice were administered the same volume of physiological saline(control roup).At 3,7,14,and 28 days after modeling,hematoxylin-eosin staining was performed to observe the pathomorphological changes of lung tissue.P16,CydinD1,and CDK4protein expression in lung tissue was detected by immunohistochemistry and Western blot analysis.RESULTS AND CONCLUSION:In the BLM group,typical changes of pulmonary fibrosis were observed,and P16 and CDK4protein expression in the pulmonary tissue increased with time,but CyclinD1 protein expression was decreased with time.P16and CDK4 protein-positive calls in the BLM group were more than in the control group.Compared with the control group,P16protein expression in the BLM group was higher at 14 and 28 days after modeling(P<0.01)and CDK4 protein expression was higher at 7,14 and 28 days after modeling(P<0.05).CydinD1 protein-positive cells and protein expression were greater at 3and 7 days after modeling in the BLM group(P<0,05),but they were less at 28 days after modeling(P<0.05),than in the control group.At 14 days after modeling,CydinD1 protein-positive cells in the BLM group were more,but CyclinD1 protein expression was ess,than in the control group(P<0.05).These findings suggest that P16,CyclinD1 and CDK4 protein expression was abnormal during pulmonary fibrosis and P16-CyclinD1/CDK4 cell cycle regulatory pathway greatly results in progressive reduction of alveolar epithelial calls during this process.

Objective To construct DNA vaccine expressing HIV-1 AE2f gp145-tat-rev-nef fusion gene( AE-Gp145TRN) and to compare the immunogenicities of DNA vaccines expressing Tat, Rev and Nef in gene fusion formulations of tat-rev-integrase(c-half)-vif-nef( AE-TRIVN) and AE-Gpl45TRN. Methods DNA vaccine was constructed by inserting the codon optimized HIV-1 AE2( gp145-tat-rev-nef fusion gene into mammalian expression DNA vector. In vitro expression efficiency of the constructed DNA vaccine was determined by Western blot and the immunogenicities of AE-Gpl45TRN and AE-TRIVN were compared by immunizing female BALB/c mice. IFN-r ELISPOT assay was used to read out the specific T cell immunity. Results Western blot assay showed the constructed DNA vaccine could be expressed efficiently in vitro. After vaccination, AE-TRIVN mounted significantly higher T cell responses against Tat, Rev and Nef[(148±91)SFCs/106 splenocytes]than Gpl45TRN[(55±28) SFCs/106 splenocytes]. Specific T cell responses elicited by AE-TRIVN predominantly targeting Rev, whereas Gpl45TRN could significantly enhance T cell responses against Nef. Conclusion AE-TRIVN and Gpl45TRN induced distinct T cell response modalities, which implied different gene fusion formulations may affect the immunogenicity of specific DNA vaccines.

Objective To prospectively study the minimally invasive hematoma stereotactic aspiration and the recovery of neurological deficits after conservative medical treatment alone, as well as the incidence of post-stroke depression (PSD) in patients with intracerebral hemorrhage, so as to investigate the effect of minimally invasive hernatorna stereotactic aspiration on the recovery of neurological deficits and the incidence of PSD. Methods Fifty-five patients with intracerebral hemorrhage received minimally invasive hematoma stereotactic aspiration (n =25) and conservative medical treatment (n = 30), respectively. The neurological deficits of the patients were assessed by the National Institutes of Health Stroke Scale (NIHSS) at admission, day 14 and 90. The decreased values (all compared to baseline score) of the NIHSS scores were calculated at day 14 and 90, respectively; the modified Rankin Scale (mRS) was used to assess the disabled degree at day 90; the 17-item Hamilton Depression Rating Scale (HAMD) was used to assess the PDS status at day 14 and 90. The correlation between PSD and the degrees of neurological deficit and disability in patients was analyzed. Results The decreased value of the NIHSS score in the stereotactic group at day 14 and 90 was significantly higher than that in the conservative treatment group (all P ＜0. 05), and the value of the mRS score was significantly lover than that in the conservative treatment group at day90 (P＜0. 05). The incidence and the total incidence of PSD in the stereotactic group at day 90 were significantly lover than those in the conservative treatment group (all P ＜0.05). There were significant positive correlation between HAMD and NIHSS scores and HAMD and mRS scores. Conclusions The recovery of neurological deficits was faster after the minimally invasive hematoma stereotactic aspiration. The degree of disability in patients was lower, and the incidence of PSD was also lower than that in the conservative treatment group. PSD was closely correlated with the degrees of neurological deficits and disability.

Objective To analysis the β-globin gene mutation in β-thalassemia in the population of Wenzhou natives,and identify the major mutation in Wenzhou and further provide valuable information for genetic counseling,prenatal diagnosis and prevention programs in this region.Methods Patients with β-thalassemia were diagnosed and the genomics DNA were extracted from whole blood cells and amplified with PCR,sequenced and compared to the standard sequence.Some mutations were further identified by subcloned.Results 44 of 66 patients were diagnosed β-Thalassemia,9 mutations were found in the 44 sporadic patients with the sequence analysis,2 of which were known polymorphisms(exonl 59,IVS-2-665),3 belonged to the common mutations in Chinese(IVS-2-654,CD_(41/42)-TTCT and TATA box nt-28),2 were scarce abnormalities(CD_(47),CD_(66))and 2 novel variants(-24T→C,CD_(26A)→G,same sense mutation,unreported).Conclusion The mutations of β-globin gene in Han Chinese in Wenzhou are complex (9 mutations found in all),the rare and novel mutations are identified,which provide the valuable information for genetic counseling in Wenzhou.

Objoctive:To investigate the expression of transforming growth factor-β(TGF-β)isoforms in colonic mucosa of ulcerative colitis(UC),and its role in the incidence of UC thereof.Methods:The expression of TGF-β isoforms was detected in colonic mucosa by immunohistochemical Envision method in UC patients,and the patients with irritable bowel syndrome (IBS)were used as controls.The relationship was analyzed between the expression of TGF-β isoforms and the degree of severity and extent of the disease.Results:There were statistical differences in the expression of TGF-β1 and TGF-β2 between the active stage UC,remission stage UC and IBS groups(P ＜ 0.01).The expression was significantly higher in active stage UC group than that in remission stage UC and 1BS groups(P＜ 0.01).The expression was significantly higher in remission stage UC group than that in IBS group(P ＜ 0.01).However,there was no statistical difference in the expression of TGF-β3 between active stage UC,remission stage UC and IBS groups(P＞ 0.05).The expression of TGF-β1 and TGF-β2 had positive correlation with the degree of sevefity(P ＜ 0.05).However,the expression of TGF-β3 had no linear correlation with the degree of severity(P ＞ 0.05).There was no linear correlation on the expressions of TGF-β1,TGF-β2 and TGF-β3 with extent of the disease(P ＞ 0.05).The degree of severity had positive correlation with extent of the disease(P ＜ 0.05).Conclusion:The expressions of TGF-β1 and TGF-β2 were enhanced in colonic mucosa of UC,and were correlated with the pathogenesis of UC.The expressions of TGFβ1 and TGF-β2 may serve as markers for assessing disease activity of UC.

Objective To investigate the effect and significance of early coronary artery bypass grafting (CABG) on the expression level of nerve-related factor at infracted border zone (IBZ) in dogs with acute myocardial infarction (AMI).Methods The anterior descending coronary artery of all thirty dogs randomly assigned into experimental group(n =22) and control group(n =8)were ligated into MI model.The experimental group included those undergoing CABG 1(n =6),2 week(n =4),4 week(n =6)and 6 week (n =6) after MI,and control group(n =2) was established for every experimental group.We detected the local expression level of nerve-related factor such as GAP43 mRNA,NGF mRNA and SYN mRNA at normal myocardium and infracted border zone myocavdium by means of RT-PCR through thoracotomy at eight weeks after surgery.Results Four dogs of every experimental group and all dogs of control group survived to the end of the study.The local expression level of GAP43 mRNA,NGF mRNA and SYN mRNA in control groups was significantly higher than that in normal and experimental groups (P ＜0.01 ).The local expression level of GAP43 mRNA,NGF mRNA and SYN mRNA in 4 and 6 weeks bypass groups was significantly higher than that in normal and 1,2 weeks bypass groups (P ＜0.05).There was no statistically significant the expression level of SYN mRNA among all experimental groups (P ＞ 0.05 ).Conclusions Early CABG surgery for AMI in dogs could lessen the expression level of nerve-related factor and the sympathetic remodelling at IBZ.Especially CABG surgery two weeks after MI could lessen the sympathetic maximacily.

Objective To establish a novel method for the determination of betaine in serum by high performance liquid chromatography with UV detection.Methods Pre-column derivatization of serum was performed directly in acetonitrile without extraction with p-bromophenacyl bromide and 18-crown-6 ether as catalyst.The p-bromophenacyl ester derivatives were then separated by Supelcosil LC-SCX, using an was 0.8 ml/min and the effluent was monitored at 259 nm. Betaine was used for preparation of standard curve and quantification with external standards.Results The linearity of this method was 6.25-200.00 μmol/L,the regression was 0.999 8.The detection limit was 3.0 μmol/L.The within-day imprecision was 1.88%-3.79% ( average 3.24% ), the between-day imprecision was 3.14%-6.76% ( average 4.39% ), the recovery rates were 95.89%-102.86% (average 99.16% ).Conclusion This method is sensitive, rapid, accurate and suitable for the research and routine clinical practice.

Objective To discuss the diagnosis,treatment and following-up of cervical intraepithelial neoplasia (CIN) during pregnancy.Methods Eighteen pregnant women with CIN,presented to the hospital from Jan.2004 to May.2008,were retrospectively reviewed.Pap smear,HPV,copolscopy and cervical biopsy were performed for diagnosis and all were followed up by Pap smear and copolscopy during pregnancy every 12 weeks,while 15 were followed up until 6-8 weeks postpartum.Results The incidence of CIN during pregnancy was 0.16%.All of the 18 women had abnormal Pap smear,followed by colopscopy and biopsy.Conservative management during the pregnancies every 3 months showed no progress in CIN.Among the 18 cases,17 delivered at term and only 1 preterm (36+4 weeks).Two out of the 18 women had normal delivery (11.1%),2 by forceps(11.1 %) and 14 by cesarean sections (77.8%).No neonatal apnea or neonatal respiratory distress syndrome had been observed.Pathological diagnosis was confirmed in all cases prenatally,including 4 CIN1,4 CIN2 and 10 CIN3.Four out of the 10 CIN3 remained after delivery,while 1 changed to CIN2.These 5 cases underwent leep electrical excision procedure and Pap smear results turned to normal at 6 to 11 months after delivery.One HSIL case were lost.The rest 4 CIN3 and all of the 8 CIN1 and CIN2 cases turned to normal or lesser degree.Fourteen of the 18 women received HPV test during pregnancy,and 12 (85.7%) were positive,among which 7 women were followed up at postpartum and 6 of them were HPV positive.Conclusions The three steps method for diagnosis of CIN during pregnancy is safe and effective.Conservative management of CIN during pregnancy is recommended.

Objective To investigate the characters of morphology change and protein expres-sion in progressed renal cell carcinoma after the treatment of sorafinib. Methods Clinical data of 2 cases with progressed renal cell carcinoma treated with sorafenib were collected. The HE slices were reviewed. Immunochemistry was used to detect the expression of Vimentin, AE1/AE3, CK7, CK8, CK18, CD10, VEGF, VEGFR2, p53 and Ki-67 levels. Results There was no difference in patho-logic type between before and after the therapy of sorafenib. Both of the 2 cases were showed degener-ation in tumor cell in different degree with fibrosis and necrosis. The expression of renal cell carcinoma related antigens (Vimentin, AE1/AE3, CK7, CK8, CK18 and CD10) had no difference before and af-ter the treatment of sorafinib. The expressions of VEGF, VEGFR2, p53 and Ki-67 were increased, and the expression of Bcl-2 was decreased after the therapy of sorafinib. Conclusions There may be some morphologie differences between the metastatic tumor or the recurrent tumor and primary tumor because of the treatment of sorafenib. However the pathologic type is the same before and after the treatment of sorafenib. The main differences are the degeneration of the tumor cell and fibrosis after the treatment of sorafenib. The expression changes of VEGF and VEGFR2 may be related to the sor-afenib application.