AIM:To construct eukaryotic expression vector of Sirt2 and detect its expression in HEK293 cells. METHODS: Total RNA was isolated from brain tissue of a-dult SD rat. A 1 130 bp fragment containing the coding region of Sirt2 was amplified by RT-PCR and the resulting PCR product was subcloned into PMD20-T vector and se-quenced. Coding region of Sirt2 was generated with PCR by using the PMD20-T-Sirt2 as template, the amplified PCR fragment was inserted into the EcoR I and Hind Ⅲ sites of the pcDNA3. 1myc-his(-)A expression vector, and the sequence was confirmed by DNA sequencing. The expression of new construct pcDNA3.1 myc-his(-) A-Sirt2 in HEK293 cells was detected by immunofluorescence. RESULTS: The full length coding region of Sirt2 was obtained and confirmed by sequencing, the expression of Sirt2 was detected successfully in HEK293 cells. CONCLUSION: The eukaryotic expression vector of Sirt2 has been successfully constructed, which will provide a useful tool for designing an in-depth investigation of the role of Sirt2.

Objective To study the effects of carboxymethylated chitosan (CMCS) to nitric oxide (NO)-induced apoptosis on rat chondrocytes,and explore p38MAPK signal transduction pathway in the process and its mechanism.Methods The rat articular cartilage cells were cultured in vitro,collagen type-2 (collagen-2) immunohistochemical staining was used to identify the cartilage cells.The model of chondrocyte apoptosis was built by different concentrations of sodium nitroprusside (SNP) induction.The cells were divided into the control group,the SNP treated group SNP+CMCS treated group,and the SNP+p38 MAPK inhibitor SB203580 treated group.The apoptotic rate of chondrocytes was calculated by FCM,apoptotic nuclei was identified by Hoechst33342 stain,the mitochondrial membrane potential changes was detected by Rhodamine123 (Rho123) stain,the expression of p38 and p-p38 were detected by Western blotting analysis.Results 1-3 mmol/L SNP could induce chondrocyte apoptosis,the apoptotic rate was increased with the SNP increasing,the most obvious apoptosis was occurred in 3 mmol/L SNP treated chondrocytes,which was 69.8％ (P＜0.05).SNP could increase the nuclear fragmentation of chondrocytes,the cells with nuclear fragmentation was significantly higher than that in the control group.SNP could reduce mitochondrial membrane potential in chondrocytes,which decreased significantly compared with the control group.SNP could increase the p-p38 expression in chondrocytes,which was 4.3 times compared to the control group.CMCS of different concentrations could reduce the apoptotic rate of SNP-induced chondrocytes,which was 51.0％,29.9％ and 15.2％,which was decreased significantly (P＜0.05) when compared with 3 mmol/L SNP induced group,CMCS decreased the cells number of SNP-induced nuclear fragmentation.CMCS increased the mitochondrial membrane potential in SNP-induced chondrocytes.CMCS reduced the expression levels of p-p38 in SNP-induced chondrocytes.Conclusion CMCS has protective effect on SNP-induced apoptosis of chondrocytes.This process is completed by inhibiting the activity of p38 MAPK signal pathway.

Humans ; Infant Nutritional Physiological Phenomena ; standards ; Infant, Newborn ; Nutritional Support ; standards ; Practice Guidelines as Topic

Animals ; Animals, Newborn ; Female ; Hyperbaric Oxygenation ; Hypoxia-Ischemia, Brain ; physiopathology ; therapy ; Nerve Growth Factor ; pharmacology ; Nerve Regeneration ; drug effects ; physiology ; Rats ; Rats, Wistar

Objective To study the damage to striatum in patients perorally addicted to compound codeine phosphate solution by using the brain dopamine transporter SPECT imaging. Methods Patients p erorally addicted to compound codeine phosphate solution ( n = 29 ) and addicted to heroin ( n = 27 ), as well as healthy volunteers (n = 31 ) were included in the study. Each of them underwent dopamine transporter (DAT) SPECT imaging with 99Tcm-2β-[N, N'-bis-( 2- mercaptoethyl ) ethylenediamino] methyl, 3β-(4-chlorophenyl)tropane (99Tcm-TRODAT-1). The striatum volume (V, cm3), mass (m, g) and radiactivity ratio (Ra) of striatum to whole brain were calculated using physio-mathematical modeling method.R esults Bilateral striatum of healthy volunteers showed typical "panda eyes" pattern and the distribution of DAT was uniform and symmetrical. Bilateral striatum of patients addicted to compound codeine phosphate showed impaired tracer uptake, similar to those addicted to heroin. The V, m and Ra of bilateral striatum of patients addicted to compound codeine phosphate were (23.68 ±4.94) cm3, (24.87 ±5.19) g and (5.01 ±0. 88 ) %, respectively, which were significantly lower than those of healthy controls: ( 35.39 ± 4.42 ) cm3,(37.16 ±4.64) g and (7.93 ±0.86)% (t = -9.69, -9.69, - 13.01, all P =0.000), but significantly higher than those addicted to heroin: ( 18.87 ± 4.66 ) cm3, ( 19.81 ± 4.90 ) g and (4.26 ± 1.02 ) % ( t =3.74, 3.74, 2.96, P = 0.000, 0.000, 0.005 ). Conclusion Long-term peroral intake of compound codeine phosphate solution may damage the function of cerebral striatum, which is someway similar to though less severe than, the impairment caused by heroin.

12E7 Antigen ; Adnexa Uteri ; surgery ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Cell Adhesion Molecules ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Granulosa Cell Tumor ; drug therapy ; metabolism ; pathology ; surgery ; Humans ; Hysterectomy ; methods ; Inhibins ; metabolism ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Ovarian Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Survival Rate ; Vimentin ; metabolism

Objective To explore clinical application of single utility port or single port video-assisted thoracoscopic lobectomy.Methods Conducting a prospective study to 191 patients with lung disease who underwent complete video-assisted thoracoscopic surgery (VATS) lobectomy in Department of Thoracic Surgery of The People's Hospital of Wuhan University from June 2013 to Dec 2014.Results Of the 191 patients, 35 underwent left upper lobectomy, 42 underwent left lower lobectomy, 43 underwent right upper lobectomy, 11 underwent right middle lobectomy, 47 underwent right lower lobectomy, and 13 underwent bilobectomy.Operations were successful in all patients with 3 patients transferred for open thoracotomy for severe adhesion or bleeding.A total of 7 of 78 upper lobectomy and 8 of 113 middle or lower lobectomy was done with adding another incision for severe adhesion.The mean operative time was 50 ～ 190 (80.3 ±43.2) min.The mean blood losing was 90 ～ 350 (145.4 ± 56.2) ml.Thirty nine patients underwent upper lobectomy, who were placed two chest tubes, respectively.The upper chest drainage duration was (1.5 ±0.8) d, and the lower chest drainage duration was (4.2 ± 1.3) d.Forty eight patients underwent lower lobectomy, middle lobectomy, or bilobectomy, who were placed one chest tube, respectively.The chest drainage duration was(4.0 ± 1.7)d.The mean recovery time after operation was (6.1 ± 2.5) d.Seventy six patients were diagnosed with lung cancer, and the average number of dissected lymph nodes from each patient was (14.7 ±6.9).The lung cancer was classified as tumor node metastasis (TNM) stage Ⅰ A, Ⅰ B,ⅡA, ⅡB and ⅢA in 67, 61, 34, 22 and 7 cases, respectively.No serious complications, such as bronchopleural fistula or death, occurred in perioperation.Conclusions In consideration of placing two chest tubes after upper lobectomy and placing one chest tube after lower lobectomy, middle lobectomy, or middle and lower lobectomy, we think single utility port-VATS (2-port) for upper lobectomy and single port-VATS lobectomy for lower lobectomy, middle lobectomy, or middle and lower lobectomy are technically safe and have the advantages of drainage, lessening pain, rapid postoperative recovery, and have no significant difference in operation time, the incidence of complications and the number of removed lymph nodes, compared to traditional 3-port-VATS.

Objective · To investigate the difference in myocardial strain of left ventricle between obstructive hypertrophy cardiomyopathy (HCM) and nonobstructive HCM. Methods · Cardiac magnetic resonance imaging (MRI) exam was performed on 48 sequential enrolled patients with HCM (18 with obstructive HCM, and 30 with nonobstructive HCM), whose left ventricular ejection fractions (LVEF) were over 50%. Twenty-five healthy volunteers were examined as normal controls. Global longitudinal strain (GLS), global radial strain (GRS), global circumferentialstrain (GCS), LVEF, left ventricular end diastolic volume (LVEDV), left ventricularmass (LVM), left ventricular end diastolic volume index (LVEDVI), and left ventricular mass index (LVMI) were collected and compared. Radial strain, circumferential strain and peak radial displacement were also measured in medial segment of left ventricle according to American Heart Association (AHA) 17-segment model. Results · ① LVEF of the patients with obstructive HCM was bigger than those of nonobstructive HCM patients and control group (P<0.05). LVM and LVMI of the HCM groups were bigger than those of control group (P<0.01). ② Left ventricle GLS, GRS, and GCS significantly decreased in the patients with nonobstructive HCM compared to those with obstructive HCM (P<0.05). The three parameters of two HCM groups were significantly lower than those of healthy volunteers (P<0.05). ③ Compared with obstructive HCM patients,the segmental parameters of left ventricule, the medial segment circumferential strain and radial strain of nonobstructive HCM patients significantly decreased (P<0.05), and the two parameters of both HCM groups were lower than those in healthy volunteers. Compared with obstructive HCM patients and healthy volunteers, peak radial displacement of left ventricule medial segment in nonobstructive HCM witnessed a significant decrease, while no significant difference was observed between obstructive HCM patients and healthy volunteers. Conclusion · In the LVEF preserved HCM patients, the myocardial strain of left ventricle in nonobstructive HCM patients decrease significantly than that in obstructive HCM patients, which may result in the different clinical outcomes intwo types of HCM patients. It is suggested that the myocardial strain is more sensitive than ejection fraction in the evaluation of myocardial performance of HCM patients.

AIM: To investigate the biological characteristics of cytokine-induced killer (CIK) cells in vitro METHODS: The non-adhere peripheral blood monoclear cells from healthy donors were induced into CIK cells in the presence of IFN-?, IL-1?, IL-2 and anti-CD3 antibody. LAK (lymphokine activated killer) cells were prepared as a control. The cellular phenotype were detected by FCM and immunocytochemistry and the cytotoxicity was measured by LDH release assay. RESULTS: After 2 weeks of induction, the proliferation rate of CIK cells reached a peak and the proportion of CD3 + population was above 95%, and then the cells growth entered to plateau phase at week 3. The proportion of CD3 +CD56 + NKT subset cells was 16 5% on day 15 and it had no obvious variety between 2 and 4 weeks. Correspondingly, LAK cells grew slowly and had lower proliferation rate compared with the CIK cells ( P

Objective To investigate the protective effects of carboxymethylated chitosan (CMCS) on nitric oxide (NO) induced apoptosis in rat chondrocytes, and the probable roles and mechanisms of PI3K/Akt signaling pathway in these process.Methods Rat knee articular cartilage was used as the source of chondrocytes, the cells were identified by immunohistochemical staining against collagen type Ⅱ, odium nitroprusside (SNP, 3 mmol/L) was used to establish the apoptotic models of chondrocytes.Cells were divided into four groups: the control group, the SNP-induced group, the SNP+CMCS treated group, the SNP+CMCS+PI3K inhibitor Wortmannin treated group.Cell proliferation were assessed by cell proliferation assay kit (CCK-8), the apoptotic rate of chondrocytes was determined by FCM with Annexin V-FITC/PI double staining, the expression levels of MMP-13 and TIMP-1 mRNA were detected by real-time polymerase chain reaction (PCR) analysis, the expression of Akt and p-Akt protein levels was detected by Western blotting analysis.One-way analysis of variance (ANOVA) statistical analysis was used to calculate the data.Results Three mmol/L SNP can inhibit proliferation (0.221±0.023), and the proliferation was reduced by 70％ compared with the control group (0.736±0.032, F=8.203, P=0.021);and the induced apoptosis in cultured chondrocytes could be observed.The apoptotic rate was (68.8±5.2)％.Increased MMP-3 and decreased TIMP-1 mRNA expression were observed in SNP-induced cells.After adding CMCS to SNP-induced chondrocytes, the proliferation was increased while apoptotic rate was decreased, the apoptotic rate decreased to (14.7±2.3)％.CMCS could promote the activation of p-Akt in SNP-induced chondrocytes and restore SNP-induced MMP-13 and TIMP-1 mRNA expression.Conclusion CMCS could protect apoptosis in SNP induced chon-drocvtes via activation of PI3K/Akt pathwav.