BACKGROUND:Construction of tissue engineering adult cardiac myocytes has been a new research hotspot in cardiovascular fields.
OBJECTIVE:To explore a simple, fast method for the separation of adult rat cardiomyocytes, and preliminarily explore the construction methods of tissue engineering adult cardiac myocytes.
METHODS:Segmented enzymatic digestion method was used to isolate cardiac myocytes from adult rats. Subsequently, cardiac myocytes were transfected with adenovirus and liposome-mediated red fluorescent protein gene. Construction efficiency of tissue engineering cells was qualified using inverted fluorescence microscopy and flow cytometry. Final y, cardiac myocytes were transfected with adenovirus-mediated hypoxia-inducible factor-1a, and the expression of hypoxia-inducible factor-1a was examined by western blot analysis.
RESULTS AND CONCLUSION:A lot of cardiac myocytes were col ected using the segmented enzyme digestion method. Flow cytometric analysis showed that the survival of adult cardiomyocytes was (87.03±0.70)%. Compared with liposomal transfection (transfection efficiency was 0), adenovirus infection efficiency was (70.31± 1.39)%, and the cells expressed red fluorescence under fluorescence microscope. After 4 days of adenovirus infection, transfected cells expressed hypoxia-inducible factor-1a protein. These results showed that segmented enzyme digestion is a fast way to isolate adult cardiac myocytes, and recombinant adenovirus vector is a good vector to transfect cardiac myocytes.

Objective To explore the effect of psychotherapy in patients with methadone maintenance treatment.Methods 60 patients accepted drug and 69 patients accepted drug with psychotherapy treatment were evaluated with Self-Rating Anxiety Scale(SAS) and Self-rating Depression Scale(SDS) in the beginning of psychotherapy therapy and 3 months later.All of them were assessed for heroin use at 3 months after the end of a 3-month treatment period.Results SAS、SDS did not show any significant differences between the two groups in the beginning(P ＞0.05).After 3 months of psychotherapy,the differences in SDS and SAS between the two groups showed statistically significant(t =- 2.858,- 2.465,all P ＜0.05).After 2 months of psychotherapy,the number of relapsing again patients of group psychotherapy treatment was significantly lower than control group(x2 =6.906,5.054,5.100,4.247,all P ＜0.05).Conclusion The mental healthy status of patients with heroin dependence is poor.Psychological treatment plays an important role in short-term prevent hernoin addicts from relapse again.

Objective The experiment aims at probing the best condition of the isolation and purification of rat islets. Methods The islets were isolated from rat pancreas by hepatopancreatic duct perfusing with collagenase and purified with Ficoll 400 discontinuous density gradient centrifugation.Then the purified islets were subjected to histological staining,electron microscopy and radioimmunoassay for identification of specificity and viability. Results The histological staining revealed that the viability and the purity of the purified islets were above 95% and 85% respectively.Electron microscopy showed that the purified islets were morphologically intact with clear membrane and plenty of secreting granules.Radioimmunoassay demonstrated the secreted insulin concerntration between low-glucose groups and high-glucose groups varied significantly,which verified the good function of the islets.Conclusion Hepatopancreatic duct perfusing with collagenase is a good method for digestion.There are many factors that influence the quantity and quality of the acquired islets,such as the completed expansion of pancreas,the concentration and viability of collagenase and the digested time,etc.

To study the clinicopathological and immunohistochemical characteristics of omental and mesenteric stromal tumor, 8 cases of stromal tumor were investigated, and followed up. The results showed that these tumors were solitary nodule, most were large. Histologically, cytoplasmic eosinophilia and perinuclear vacuolization were often seen in omental and mesenteric stromal tumor of spindle cell type. 3 cases showed epithelioid cytologic features. An immunohistochemical study showed that all the patients were C kit positive (100%),7 were CD34 positive, whereas 6 were SMA positive, positive expression of p53 and PCNA was found in 2 cases of malignant stromal tumor, Lower expression of PCNA was observed in 1 case of benign one.The results suggested that omental and mesenteric stromal tumors have distinct clinicopathological features, the expression of C kit and CD34 plays an important role in its diagnosis.

Objective To observe the pulmonary X-ray changes of malignant malaria.Methods The chest routine X-ray examination in 86 cases with malaria in company with respiratory tract symptoms such as fever was performed,the pulmonary changes were obsenved.Results Of 86 cases, 58 cases were abnormal on chest filmes,including lung framework marked in 33 cases,miliary,small or large patchy shadow in 24 cases and multiple round opaque mass lesion in one case.Conclusion The pulmonary X-ray changes of malignant malaria are varied,the common characteristic is that most of them are situated at middle and lower portion of the lungs.This may attribute to blood dynamic abnormality resulting from malaria protozoan and the pathological changes of involved tissues.

Objective The experiment aims at probing the best condition of the isolation and purification of rat islets.Methods The islets were isolated from rat pancreas by hepatopancreatic duct perfusing with collagenase and purified with Ficoll 400 discontinuous density gradient centrifugation.Then the purified islets were subjected to histological staining,electron microscopy and radioimmunoassay for identification of specificity and viability.Results The histological staining revealed that the viability and the purity of the purified islets were above 95％and 85％respectively.Electron microscopy showed that the purified islets were morphologically intact with clear membrane and plenty of secreting granules.Radioimmunoassay demonstrated the secreted insulin concerntration between low-glucose groups and high-glucose groups varied significantly,which verified the good function of the islets.Conclusion Hepatopancreatic duct perfusing with collagenase is a good method for digestion.There aye many factors that influence the quantity and quality of the acquired islets,such as the completed expansion of pancreas,the concentration and viability of collagenase and the digested time,etc.

0.05).Conclusion The SIS method not only can determine multiple index in Chinese materia medica(CMM)simultaneously,but also be simple and accurate.It will be a good quality evaluation pattern for CMM.

OBJECTIVE: To measure the upper airway of obstructive sleep apnea-hypopnea syndrome by speed CT. We can predict the airway obstruction plane with the airway plane data and compliance in OSAHS patients. Through this measurement, we can provide assistance for clinical diagnosis and treatment. METHOD: This study randomly selected 82 patients diagnosed with OSAHS and 45 cases non-snoring healthy people as control group by the PHILIPS 256-slice CT. The zone volume, sagittal diameter, coronary diameter and cross-sectional area of the narrowest plane in nasopharyngeal area, velopharyngeal area, glossopharyngeal area, hypopharynx area in two groups of quiet respiration and Müller's status were measured. RESULT: In the OSAHS group underwent quiet breathing and Müller movement during CT scanning, the two states about nasopharyngeal coronary diameter, the hypopharynx sagittal diameter and cross-sectional area showed no significant difference, while other groups showed differences between the parameters (P<0.05). There was significant difference (P<0.01) in the velopharyngeal volume, tongue sagittal diameter and volume. Under Müller movement in the OSAHS group and the control group, the hypopharynx volume showed no significant difference. The nasopharyngeal coronary diameter and volume, velopharyngeal cross-sectional area and tongue sagittal diameter were different (P<0.05). The remaining set of parameters showed significant differences (P<0.01). Compared the airway compliance (Müller phase) of the OSAHS group and the control group,the parameters of each group were different (P<0.05), of which the nasopharyngeal sagittal diameter, velopharyngeal volume showed significant difference (P<0.01). Meanwhile, in the same plane, coronary diameter was greater than sagittal diameter (P<0.05) in both the OSAHS group and the control group. The pharyngeal volume measurement was basically consistent to the fiber endoscopy. CONCLUSION The obstruction plate of OSAHS patients is mostly in the velopharyngeal area and glossopharyngeal area measured with a high speed CT. The volume measurement of upper airway with speed CT can predict airway obstruction plate in patients with OSAHS.
Case-Control Studies ; Humans ; Hypopharynx ; Nasopharynx ; diagnostic imaging ; Pharynx ; diagnostic imaging ; Sleep Apnea, Obstructive ; diagnostic imaging ; Tomography, X-Ray Computed ; Tongue ; diagnostic imaging

Objective:To construct the recombinant eukaryotic expression vector plasmids with mutant C-kit cDNA and to study the effect of the mutant C-kit gene on cell proliferation and cell cycle in gastrointestinal stromal tumor (GIST). Methods: Wild-type C-kit cDNA was cloned from human embryonic brain tissue by RT-PCR technique.Site-directed mutagenesis of the wild type C-kit cDNA was performed according to the C-kit mutations we cloned. Recombinant plasmids were stably transfected into human embryonic kidney cell line and the cells expressing mutant C-kit were selected by special cell culture medium containing G418.Expressions of C-kit protein of the transfectants were detected by Western blot.Cell proliferation and cell cycle of the transfectants were detected by MTT clolorimetic assay and flow cytometry,respectively.Whether HEK cell with mutated C-kit cDNA could grow autonomously in nude mice or not was also detected. pcDNA3 vector transfected and recombinant plasmids with wild-type C-kit cDNA transfected HEK cell were used as the control groups. Results: The mutant C-kit cDNA was obtained by site-directed mutagenesis of the wild type C-kit cDNA. Compared with the 2 control groups ,the growth rate and proliferative activity of the HEK cells with mutant C-kit cDNA were increased significantly.The analysis of cell cycle showed that more HEK cells with mutanted C-kit cDNA remained in proliferation phase (S+G 2-M)than the groups without mutated C-kit cDNA.HEK cells with the mutated C-kit also grew autonomously in nude mice.Conclusion: Mutation of C-kit gene can increase proliferation of human cells,causing malignant transformation of human normal cells,which may play an important role in the malignant transformation of GIST.

Objective: To explore the sensitivity of Kit or PDGFRA mutants related to gastrointestinal stromal tumor (GIST) to Gleevec.Methods: The recombinant plasmids of KIT Del559-560, KIT Ins IPYD579, PDGFRA D842V and PDG-FRA L839P gene mutants were transiently transformed into the CHO cells by liposome methods.Western blot was used to detect the expression of the related protein and their phosphorylated forms after the cells were incubated with Gleevec for 90 min.At 72 hours after incubation with Gleevec, MTT was used to detect cell proliferation.Results: Western blot results showed that Gleevec at 0.1 μM can notably reduce phosphorylation of KIT Del559-560.Gleevec at 1μM completely blocked phosphorylation of KIT Ins IPYD579 and PDGFRA L839P, but did not affect PDGFRA D842V phosphorylation.MTT analy-sis indicated that growth of CHOPDGFRA L839P was inhibited by Gleevec at 1μM, however, CHOPDGFRA D842V was re-sistant to Gleevec at 5 μM.Conclusion: Gleevec can decrease the expression of phosphorylated protein CHOPDGFRA L839P and CHOKIT Ins IPYD579, and can remarkably inhibit the proliferation of cells containing PDGFRA L839P mutant.