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Objective: To investigate the role of occludin in tight junction (TJ) in vitro., Methods: We constructed RNA interfering lentiviral vectors and transfected them into TM4 cells. Then we detected their inhibitory effect on occuldin by RT-PCR and Western blot and analyzed the role of occuldin in TJ using an in vitro TJ cell model., Results: The pLenti 6.3-EGFP-occludin-miR expression vector was successfully constructed. The results of RT-PCR and Western blot showed that pLenti 6.3-EGFP-occludin-miR-3 significantly inhibited the expression of occludin (P < 0.05), which was remarkably lower than in the blank control and the pLenti 6.3- EGFP transfection group (0.7534 ± 0.089 vs 1.000 and 1.056 ± 0.025, P < 0.05). The expression of occludin was markedly suppressed and the tightness of tight junctions decreased in the TM4 cells transfected with pLenti 6.3-EGFP-occludin-miR-3., Conclusions: The pLenti 6.3-EGFP-occludin-miR expression vector was successfully constructed, and occludin is one of the functional proteins that maintain tight junctions.

Objective: To study the transcription factors of the spermatogenesis-related promoter mir-122-5p., Methods: SP1 and GATA4 were predicted as the possible transcription factors of the mir-122-5p promoter by bioinformatics analysis, followed by construction of the double luciferase pGL3-mir-122-5p promoter vector, pcDNA3.1 (+) -SP1 expression vector and pcDNA3.1 (+) -GATA4 expression vector, respectively. The pcDNA-SP1+pGL3-basic mixture plasmid and pcDNA-SP1+ pGL3-miR-122-5p promoter mixture plasmid, pcDNA-GATA4+pGL3-basic mixture plasmid and pcDNA-GATA4+pGL3-miR-122-5p promoter mixture plasmid were transferred into 293T cells. The enzyme activity was detected the Dual-Luciferase Reporter Assay System., Results: The fluorescence value of the pcDNA3.1+pGL3-miR-122 promoter was 0.0362 ± 0.0004, significantly higher than that of the pcDNA3.1+pGL3-basic group (P < 0.05), indicating the successful construction of the mouse miR-122-5p promoter luciferase reporter plasmid. The fluorescence value was markedly higher in the pcDNA -SP1 + pGL3-miR-122-5p promoter than in the pcDNA -SP1+pGL3-basic group, suggesting that the transcription factor SP1 could promote the transcription of miR-122. There was no statistically significant difference in the fluorescence value between the pcDNA -gata4+pGL3-basic transfection and pcDNA -GATA4+pGL3-miR-122-5p promoter transfection groups, indicative of the inability of GATA4 to promote the transcription of miR-122-5p., Conclusions: The transcription factor SP1, rather than GATA4, can promote the transcription of miR-122-5p.

Pulmonary hypertension is a life-threatening medical condition, and a growing body of evidence shows that the expression of connective tissue growth factor (CTGF) is significantly associated with its pathogenesis, making it an attractive therapeutic target. Our earlier work revealed that plasmid-based CTGF-specific short hairpin RNA (shRNA) could attenuate pulmonary artery smooth muscle cell (PASMC) proliferation and pulmonary vascular remodeling in rats exposed to cigarette smoke. In this study, we explored the therapeutic role of this shRNA plasmid in the treatment of monocrotaline-induced pulmonary vascular remodeling in rats, and demonstrated that the upregulation of CTGF in PASMCs following a single injection of monocrotaline could be attenuated by administration of the shRNA. Accordingly, this shRNA was found to repress monocrotaline-induced pulmonary vascular remodeling, as evidenced by its ability to reduce the percentage of muscularized vessels and the wall thickness of pulmonary vessels. We concluded that plasmid-based shRNA against CTGF attenuated pulmonary vascular remodeling in monocrotaline-treated rats. CTGF might be a potential target for the treatment of pulmonary vascular remodeling and pulmonary hypertension.

Objective: To investigate the expression of CXCL12, its receptor CXCR4 and its correlations with clinical pathology and lymphangiogenesis in pancreatic adenocarcinoma (PAC)., Methods: The tissue samples were obtained from 30 patients with PAC by surgery between January 2005 and December 2007, which including PAC, the cancerous peripheral tissues, the normal pancreatic tissues and peripheral lymph nodes. The patients age ranged from 35 to 78 years old (median 57.2 years old). The expressions of CXCL12 and CXCR4 in these tissues were assayed by immunohistochemical staining, RT-PCR and fluorescence quantitative real-time PCR., Results: In the immunohistochemical staining, the CXCL12 protein mainly located in the normal pancreatic cell envelopes and/or cytolymphs. In the immunohistochemical staining, the CXCR4 protein mainly located in the cell envelopes and/or cytolymphs of PAC. The results of RT-PCR and fluorescence quantitative real-time PCR indicated that the expression levels of CXCR4 mRNA in PAC tissues, the cancerous peripheral tissues and peripheral lympho nodes were higher than that in the normal pancreatic tissues (P < 0.01). The MLVD in PAC were detected by morphometric analysis respectively. The level of MLVD in III-IV stages was higher than I-II stages of PAC (P < 0.01), and in these cases which had lymphatic metastasis, the level of MLVD significantly increased (P < 0.01). And there was no correlation between the differentiation and histology types of PAC (P > 0.05). There was 22 samples that the CXCR4 protein was positive, and among these samples the MLVD was higher than that in negative group of CXCR4 protein (P = 0.003)., Conclusions: The expression of CXCR4 was significantly associated with lymphatic metastasis of PAC, and the higher expression of CXCR4 in PAC tissues was significantly associated with lymphangiogenesis of PAC.

Objective: To study the expression of Src-suppressed C kinase substrate (SSeCKS) mRNA in pulmonary microvascular endothelial cells(PMVEC) induced by lipopolysaccharide, and the interfering effect of methylprednisolone sodium succinate., Methods: Rat PMVEC (RPMVEC) were isolated and cultured in vitro, then grouped randomly according to different culture time and different dosage of LPS, and the expression of SSeCKS mRNA in RPMVEC of different groups were determined by reverse transcription polymerase chain reaction (RT-PCR)., Results: Normally, the expression of the SSeCKS mRNA in RPMVEC was maintained on a low level, and it could be elevated by stimulation of LPS, and the changes in the expression exhibited in dosage and time dependent manner. After being stimulated with LPS for 1 hour, the extent of increase in SSeCKS mRNA in RPMVEC coincided with the increased dosage of LPS (control: 0.263+/-0.033; LPS of 0.1 mg/L: 0.529+/-0.066; LPS of 1 mg/L: 1.391+/-0.048; LPS of 10 mg/L: 2.339+/-0.055; LPS of 100 mg/L: 2.861+/-0.069, F=639.096, P<0.05). When RPMVEC were stimulated with 10 mg/L of LPS, the expression of SSeCKS mRNA began to increase at 0.5 hour, peaked at 1 hour, then decreased gradually, and it remained high even at 12 hours (control: 0.301+/-0.022; LPS 0.5 hour: 1.617+/-0.018; 1 hour: 2.378+/-0.031; 3 hours: 2.148+/-0.056; 6 hours: 1.322+/-0.042; 12 hours: 0.772+/-0.044, F=726.346, P<0.05); After methylprednisolone sodium succinate had been given, the increase of SSeCKS mRNA induced by LPS is inhibited (2.664+/-0.104 vs. 1.759+/-0.151, F=156.000, P<0.05)., Conclusion: (1) LPS could induce up-regulation of SSeCKS mRNA, the elevation is in time and dosage dependent manner. This indicate SSeCKS is related with LPS induced injury of RPMVEC. (2) Methylprednisolone sodium succinate inhibit the increase of SSeCKS mRNA induced by LPS.

A multiple tracer dilution method measuring total body water, extracellular water, plasma volume, and red cell volume was used to study body composition. Healthy Chinese subjects were compared to a group of Chinese patients with various newly diagnosed gastrointestinal disorders scheduled for elective operation. Compositional data obtained from these groups showed no significant differences in compartmentalization, suggesting absence of major changes during the early stages of certain gastrointestinal diseases. In addition, healthy Chinese subjects were compared to a group of healthy Americans previously evaluated by similar dilutional methods. In males, body weight (kg) differed considerably (Chinese = 62.1 +/- 2.0 versus American = 72.5 +/- 4.1, p less than 0.02) and could be explained by alterations in body fat (Chinese = 12.4 +/- 1.3 versus American = 19.2 +/- 2.2, p less than 0.02) and extracellular water (Chinese = 14.4 +/- 0.5 versus American = 16.8 +/- 0.8, p less than 0.02). Functional tissue ("body cell mass") was similar in the 2 groups which suggested that Chinese have comparable body composition without an additional fat burden. The multiple tracer technique using deuterium oxide, sodium bromide, Evans dye, and Cr51 for body compositional assessment is accurate but expensive and laborious. Therefore, equally precise but more economical bedside methods are needed for routine compositional analysis.

Until recently, China had been isolated from Western countries for almost 30 years. Surgeons of Western countries, perhaps, are interested to know what is happening in surgery in China. This communication reviews the current interests in Chinese surgery. It is by no means complete, nor is that the intention. The intention is to introduce to our foreign colleagues a general overview of some highlights in Chinese surgery.

A variety of investigators have attempted to improve nitrogen balance during the postoperative period by modifying the composition of the infused nutrient solutions. This study compared the metabolic effects of administering standard amino acid solutions with a solution enriched with branched chain amino acids (BCAA). A prospective, randomized clinical study was performed in patients who had undergone subtotal gastrectomy or hemicolectomy, and subsequently cared for in the metabolic care unit. The patients were selected from specific entry criteria so that two groups of individuals were comparable. All patients underwent operation without complications. The plasma concentrations of valine and leucine were significantly increased (p less than 0.05 and p less than 0.01, respectively) two days after administration of solutions enriched with BCAA and throughout the entire postoperative period. The plasma glutamine concentrations tended to decrease in both groups; no concentration difference occurred between groups. Nitrogen balance tended to be more positive in the group receiving BCAA but there was no significant difference between groups after operation. Urinary excretion of 3-methylhistidine tended to increase postoperatively in both groups, but no difference occurred between groups. However, the urine excretion of isoleucine increased significantly in the patients receiving infusions enriched with BCAA. Both standard balanced amino acid and amino solutions enriched with BCAA were well tolerated in all patients.

This report is based on 31 years of experience with 116 cases of hyperinsulinism. Six cases had hypertrophy of the islets of Langerhans, 3 had widespead metastasis from malignant insulinomas, and 107 were benign adenoma cases. An immunoreactive insulin to glucose ratio of 0.3 of the peripheral venous blood before operation is of great value in diagnosing hyperinsulinism. Intraoperatively, immunoreactive insulin assay of the portal blood (IRI) is very valuable in determining if an insulinoma remains. The dividing line is 100 microU.ml-1. In localizing the tumor, "differential" PTPC is important before operation. During the operation, fine needle aspiration cytology may assist in ascertaining if the palpable tumor is an insulinoma. Multiple fine needle aspiration cytology examinations can sometimes reveal an insulinoma in an indurated pancreas. Portal vein blood IRI and blood sugar assays may serve to confirm if removal of the insulinoma is complete. Removal of the insulinoma controls hypoglycemia satisfactorily, but the brain damage incurred by prolonged hypoglycemia cannot be significantly altered. Removal of the tumor should be by enucleation, and the raw surface of the pancreas should be drained not sutured.