Establishment of a modified BISAP scoring system and its clinical significance in the early diagnosis and prognostic evaluation of severe acute pancreatitis / 临床肝胆病杂志

ObjectiveTo establish a modified BISAP scoring system， and to investigate the value of the BISAP scoring system versus the modified BISAP scoring system in assessing the severity and condition of acute pancreatitis （AP）. MethodsFor the establishment of the new scoring system， a retrospective analysis was performed for the clinical data of 1 033 patients with AP who were admitted to Third Xiangya hospital of central South University from January 2019 to December 2021， and according to the revised Atlanta classification， they were divided into mild acute pancreatitis （MAP） group with 827 patients and severe acute pancreatitis （SAP） group with 206 patients. The two groups were compared in terms of clinical features， laboratory markers， and imaging data. A binary logistic regression analysis was performed for the statistically significant indicators to screen for the independent risk factors for SAP. The receiver operating characteristic （ROC） curve was used to obtain the optimal cut－off value corresponding to the maximum Youden index for each independent risk factor， and a score of 0 or 1 was assigned depending on different situations， which was integrated into the BISAP scoring system to establish a modified BISAP scoring system. For the validation of the new scoring system， a retrospective analysis was performed for the clinical data of 473 patients with AP who were admitted to Third Xiangya hospital of central South University from January 2017 to December 2018. BISAP score and modified BISAP score were determined for each patient， and the area under the ROC curve （AUC） was used to compare the value of the two scoring systems in predicting the severity and prognosis of AP. The chi－square test or the Fisher’s exact test was used for comparison of categorical data between two groups， and the independent－samples t test and the Mann－Whitney U test were used for comparison of continuous data between two groups. ResultsFor the establishment of the new scoring system， there were significant differences between the MAP group and the SAP group in mode of admission， length of hospital stay， ICU admission rate， number of deaths， underlying diseases， and incidence rate of complications （all P<0.05）. The binary logistic regression analysis showed that body temperature， neutrophil－to－lymphocyte ratio （NLR）， C－reactive protein （CRP）， albumin， triglycerides， D－dimer， fibrinogen， and MCTSI score were independent risk factors for SAP （all P<0.05）. The ROC curve analysis showed that CRP （AUC=0.921）， NLR （AUC=0.798）， D－dimer （AUC=0.768）， and MCTSI score （AUC=0.931） had a good predictive value for SAP， and the combination of these four indicators had an AUC of 0.976 and showed a significantly higher diagnostic efficiency than each indicator alone or the combination of two or three indicators （all P<0.05）. For the validation of the new scoring system， a total of 473 patients were enrolled， with 408 in the MAP group and 65 in the SAP group， and there were significant differences between the two groups in mode of admission， length of hospital stay， ICU admission rate， number of deaths， and incidence rate of complications （all P<0.05）. The modified BISAP score was better than the BISAP score in predicting SAP （AUC： 0.972 vs 0.887， P<0.05）， with an optimal cut－off value of >3 points. The modified BISAP score also had a relatively high value in predicting the mortality of AP patients （AUC=0.910）， but there was no significant difference between the modified BISAP score and the BISAP scoring system （AUC： 0.910 vs 0.896， P=0.707）. ConclusionThe modified BISAP score is better than the BISAP score in predicting the severity of AP and has a relatively high value in predicting the mortality of AP patients， giving a more accurate， objective， and early assessment of the condition of AP patients.

A four-protein metabolon assembled by a small peptide protein creates the pentacyclic carbonate ring of aldgamycins

Organic carbonates (OCs) are a class of compounds featured by a carbonyl flanked by two alkoxy/aryloxy groups. They exist in either linear or cyclic forms, of which the majority encountered in nature adopt a pentacyclic structure. However, the enzymatic basis for pentacyclic carbonate ring formation remains elusive. Here, we reported that a four-protein metabolon (AlmUII-UV) assembled by a small peptide protein (AlmUV) appends a reactive

Short-term curative effect of superior rectal artery preserving laparoscopic resection of sigmoid colon carcinoma / 中华普通外科杂志

Objective To study the operational techniques and feasibility of superior rectal artery preserving laparoscopic resection of sigmoid colon carcinoma.Methods From Jan 2015 to Nov 2016,85 patients with sigmoid colon carcinoma were divided into artery preserving group (27 cases) and traditional surgical group (58 cases).Results The mean operation time was (283 ± 51) min,the mean lymph node dissection was (15 ± 8) and the mean blood loss was (62 ± 17) ml in the artery preserving group.The mean operation time was (179 ±e63) min,the mean lymph node dissection was (15 ±7) and the mean blood loss was (67 ± 17) ml in the traditional surgery group.The number of resected lymph nodes and blood loss had no statistical significance between these two groups (t =0.058,P ＞0.05).Longer operating time were observed in the retained vascular group as compared to the traditional surgical group (t =7.530,P ＜ 0.05).There was no anastomotic fistula in the retained vascular group,however,two anastomotic fistula cases occurred in the traditional surgical group (x2 =0.043,P ＞ 0.05).Conclusions Preservation of superior rectal artery was safe and feasible for laparoscopic resection of sigmoid colon carcinoma.

Establishes model of liver cirrhosis with portal hypertension by portal infusion of 80％ alcohol in swines / 中国介入影像与治疗学

Objective To explore the feasibility of establishing a swine model of liver cirrhosis with portal hypertension by portal infusion of 80％ alcohol.Methods A total of 13 Guizhou miniature pigs were randomly divided into three groups,experiment group 1 (n=5),experiment group 2 (n=5) and control group (n=3).Experiment groups of pigs received portal infusion of 80％ alcohol in volumes of 5 ml in group 1,and 10 ml in group 2,and the pigs in control group received portal perfusion of saline in volumes of 10 ml.All animals were performed direct portal angiography,the portal vein pressures and diameter were also detected before,immediately and 6 weeks after the infusion.All animals underwent liver biopsies before and 6 hours,1-6 weeks after operation.And contrast-enhanced abdominal CT was performed before and 6 weeks after operation.All animals were dissected 6 weeks after operation,aud each leaf of liver specimens were performed histological examination.Results There was no statistically significant difference of the portal venous pressure and diameter before infusion and 6 weeks after infusion in the experiment group 1 and control group (all P＞0.05).In the experiment group 2,compared with pre infusion,the portal vein pressure and diameter were higher than those of immediately and 6 weeks after infusion (all P＜0.05).In both experiment group 1 and group 2,all pigs had developed into liver fibrosis,the METAVIR score of 2 pigs in group 1 and 5 pigs in group 2 respectively were up to grade 4.Conclusion Portal infusion of 80％ alcohol is more suitable for establishing a swine model of liver cirrhosis with portal hypertension.

Role of portal venous pressure changes in the liver dysfunction caused by hepatic congestion after extended liver resection / 中华消化外科杂志

Objective To explore the role of portal venous pressure changes in the liver dysfunction caused by hepatic congestion after extended liver resection.Methods The experimental study was adopted.According to the random number table,90 Sprague-Dawley rats were divided into 3 groups,30 in each group:30 rats in the non-congestion group received 70％ of liver resection (median lobe + left lobe),30 rats in the congestion group received 70％ of liver resection (median lobe + left lobe) with whole caudal lobe congestion by ligation of veins and 30 rats in the congestion + splenectomy group received 70％ of liver resection (median lobe + left lobe) with whole caudal lobe congestion by ligation of veins and splenectomy.(1) Twenty rats in each group were used to make postoperative survival analysis.Ten rats in each group were used for related experiments.The portal venous pressures (PVPs) of 5 rats in each group were detected at postoperative 12 hours and 24 hours,and then blood and liver specimens were collected.(2) PVP changes were detected at postoperative 12 hours and 24 hours.(3) Clinical and biochemical test:level of total bilirubin (TBil) was tested at postoperative 12 hours and 24 hours.(4) Pathological examination:liver pathological damage was detected by HE staining.(5) The expression of CD68 macrophagocyte was detected by immunohistochemical staining.(6) The relative expressions of Cleaved Casepase-3 and hypoxia inducible factor-1α (HIF-1α) proteins at postoperative 24 hours were detected by Westein blot.(7) The relative expressions of mRNA of vascular regulation related genes (ET-1/eNOS) and inflammatory factors (TNF-α and IL-6) were detected by real-time polymerase chain reaction (RT-PCR).(8)The hyaluronic acid (HA) was measured by enzyme-linked immuno-sorbent assay (ELISA).Measurement data with normal distribution were represented as （x） ± s.Comparison among 3 groups was done using the ANOVA,and pairwise comparison was done by the LSD test.The postoperative 5-day survival curve was drawn by the KaplanMeier method,and the survival was compared using the Log-rank test.Results (1) Survival analysis:5-day survival rate in the non-congestion group,congestion group and congestion + splenectomy group were respectively 75％,10％ and 55％,with a statistically significant difference among the 3 groups (x2=18.21,P ＜0.05).(2)Changes of PVPs and TBil:levels of PVP and TBil in the non-congestion group,congestion group and congestion + splenectomy group were respectively (15.77 ±0.67)cmH2O,(18.33 ±0.28) cmH2O,(14.87 ± 0.58) cmH2O,(1.48 ±0.10)μmol/L,(1.76±0.15) μ mol/L,(1.62 ±0.11) μmol/L at postoperative 12 hours and (13.49 ± 0.45) cmH2 O,(16.96 ± 0.82) cmH2 O,(15.69 ± 0.85) cmH2 O,(1.47 ± 0.11) μmol/L,(1.94 ± 0.07) μmol/L,(1.67 ± 0.11) μmol/L at postoperative 24 hours,showing statistically significant differences among 3 groups (F =56.53,29.01,6.81,27.85,P ＜ 0.05).(3) Results of pathological examination:compared with noncongestion group,there were a lot of vacuolar cells with degeneration appearing in non-congestion liver tissues,severe liver cell swelling and hepatic sinus congestion in the congestion group at postoperative 24 hours.Compared with congestion group,vacuolar degeneration appearing in non-congestion liver tissues have some improvement in the congestion + splenectomy group.(4) Immunohistochemical staining:compared with non-congestion group and congestion + splenectomy group,the positive CD68 marked macrophages in the congestion group were increased at postoperative 24 hours.(5) Western blot assay:the relative expressions of Cleaved Casepase-3 and HIF-1α proteins in the non-congestion group,congestion group and congestion + splenectomy group were 0.63 ± 0.05,1.17 ± O.18,0.95 ± 0.17 and 0.63 ± 0.14,1.48 ± 0.08,1.13 ± 0.17,respectively,showing statistically significant differences among 3 groups (F =17.42,50.58,P ＜ 0.05).(6) Results of RT-PCR:the relative expression of mRNA of ET-1/eNOS in the non-congestion group,congestion group and congestion + splenectomy group was respectively 1.01 ± 0.63,2.09 ± 0.27,0.82 ± 0.12 at postoperative 12 hours and 0.73 ± 0.17,2.16 ± 0.94,0.80 ± 0.24 at postoperative 24 hours,showing statistically significant differences among 3 groups (F =62.91,10.65,P ＜0.05).The relative expression of mRNA of TNF-α in the non-congestion group,congestion group and congestion + splenectomy group was respectively 0.99 ± 0.08,127.80 ± 13.15,7.34 ± 1.56 at postoperative 12 hours and 0.99 ± 0.06,116.62 ± 13.32,58.62 ± 12.12 at postoperative 24 hours,showing statistically significant differences among 3 groups (F =436.77,154.54,P ＜ 0.05).The relative expression of mRNA of IL-6 in the non-congestion group,congestion group and congestion + splenectomy group was respectively 0.98 ±0.06,1.87 ±0.34,1.54 ±0.15 at postoperative 12 hours and 0.99 ±0.05,2.02 ±0.27,1.51 ±0.11at postoperative 24 hours,with statistically significant differences among 3 groups (F =22.08,46.71,P ＜ 0.05).(7) Results of ELISA:the level of HA in the non-congestion group,congestion group and congestion + splenectomy group was respectively (149 ± 9) ng/L,(200 ± 19) ng/L,(174 ± 9) ng/L at postoperative 12 hours and (136 ± 16) ng/L,(202 ± 13) ng/L,(91 ± 11) ng/L at postoperative 24 hours,with statistically significant differences among 3 groups (F =19.23,34.68,P＜0.05).Conclusions On the basis of extended liver resection,a wide range of liver congestion through increasing PVP causes hepatic microcirculation disorders,hypoxia,inflammation,vacuoles degeneration cells,increased cells apoptosis,aggravated damage of liver function and increased mortality of rats.Splenectomy could reduce PVP and then improve the liver tissues damage caused by liver congestion,meanwhile,increase the survival rate of rats.

Dynamic variation patterns of aconitum alkaloids in daughter root of Aconitum carmichaelii [Fuzi] in the decoction process based on the content changes of nine aconitum alkaloids by HPLC- MS- MS

The chemical components in the decoctions of Chinese herbal medicines are not always the same as those in the crude herbs because of the insolubility or instability of some compounds. In this work, a high-performance liquid chromatography [HPLC] coupled with electrospray ionization [ESI] tandem mass spectrometry method was developed to explore dynamic variation patterns of aconitum alkaloids in Fuzi during the process of decocting aconite root. The fragmentation patterns of aconitum alkaloids using ESI and collision-induced dissociation [CID] techniques were reported. This assay method was validated with respect to linearity [r[2] > 0.9950], precision, repeatability, and accuracy [recovery rate between 94.6 and 107.9%].The result showed that the amounts of aconitum alkaloids in the decoction at different boiling time varied significantly. In the decoction process, the diester- type alkaloids in crude aconite roots have transformed into Benzoylaconines or aconines

Knockdown of Notch-1 augments inhibitory effect of dihydroartemisinin on viability of human osteosarcoma cell line U-2OS / 中国病理生理杂志

AIM:To investigate the effect of Notch-1 knockdown on the growth of dihydroartemisinin-inhibited human osteosarcoma cell line U-2OS.METHODS:U-2OS cells treated with different concentrations of dihydroartemisinin (5, 10, 15 and 20μmol/L) were collected.The expression of Notch-1, MMP-2, MMP-9 and Hes-1 at mRNA and protein levels was measured by real-time PCR and Western blotting, respectively.U-2OS cells were transfected with Notch-1 siRNA for 24 h and incubated with dihydroartemisinin for another 24 h.The cell apoptotic rate , protein expression of MMP-2, MMP-9 and Hes-1, and the migration ability were measured by MTT assay , Western blotting and Transwell experiment , respectively.RESULTS:Dihydroartemisinin (5, 10, 15 and 20 μmol/L) decreased the expression of Notch-1, MMP-2, MMP-9 and Hes-1 at mRNA and protein levels in a dose-dependent manner .Down-regulation of Notch-1 significantly en-hanced the effect of dihydroartemisinin on the cell apoptosis , the protein expression of MMP-2, MMP-9 and Hes-1, and mi-gration ability ( P<0.05 ) .CONCLUSION: Notch-1 pathway is involved in the process of dihydroartemisinin-inhibited U-2OS cell growth.Knockdown of Notch-1 augments the inhibitory effect of dihydroartemisinin on U-2OS cell viability.

Biocompatibility of Ti35Nb3Zr2Ta, a new beta-titanium alloy, as joint prosthesis material / 中国组织工程研究

BACKGROUND:Ti6Al4V is a titanium aloy that is widely used in human joint replacement, but its modulus of elasticity is greater than human bone, resulting in the bad stability of the prosthesis. Ti35Nb3Zr2Ta, a new βtitanium aloy, has a lower modulus of elasticity, and maybe becomes a new-generation human joint prosthesis material that has a better biocompatibility. OBJECTIVE:To study the biocompatibility of Ti35Nb3Zr2Ta in prosthesis. METHODS:Wanfang, CNKI and PubMed databases were retrieved using a computer with “new β titanium;prosthesis; biocompatible” as keywords, and the retrieval time ranged from 2010 to 2015. Articles focusing on current application status for medical prosthesis materials and the biocompatibility of Ti35Nb3Zr2Ta in prosthesis were selected. RESULTS AND CONCLUSION:Compared with Ti6Al4V, Ti35Nb3Zr2Ta has higher surface roughness and smaler surface contact angle; the alkaline phosphatase activity and amount of calcium deposits in osteoblasts cultured at Ti35Nb3Zr2Ta is significantly higher than that at Ti6Al4V. Ti35Nb3Zr2Ta has good biocompatibility, and can be considered to be widely used as joint prosthesis material further.

Learning curve for middle pancreatectomy / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the learning curve for middle pancreatectomy by comparing the outcomes of middle pancreatectomy operated by a single treatment group at different stages.</p><p><b>METHODS</b>A total of 48 patients received middle pancreatectomy by single treatment group between January 2006 and April 2014 at our hospital. These 48 cases were divided into 10 stages (5 cases in each) according to the operation sequence. The operation time, blood loss, surgical complications, rate of negative margin and postoperative hospital stay were analyzed retrospectively.</p><p><b>RESULTS</b>There was no significant difference among the 10 stages in respect to surgical complications, rate of negative margin and postoperative hospital stay (P>0.05). The median operation time and blood loss in the first stage was 375 min and 530 ml, respectively. The median operation time and blood loss in the second stage was 280 min and 330 ml, respectively. There were significant differences between these two stages and the other later stages in median operation time and blood loss (P<0.01). However, there was no significant difference among the stages 3 to 10 in the median operation time and blood loss (P>0.05 for all).</p><p><b>CONCLUSION</b>After 10-15 cases of middle pancreatectomy, a surgeon can be skilled and experienced in this surgical procedure with few surgical complications.</p>

Correlation between clinicopathological features and CA19-9/CEA in patients with extrahepatic cholangiocarcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the correlation between clinicopathological features and serum carbohydrate antigen 19-9 (CA19-9)/carcinoembryonic antigen (CEA) in patients with extrahepatic cholangiocarcinoma (ECC).</p><p><b>METHODS</b>The clinicopathological data of 126 cases of extrahepatic cholangiocarcinoma treated in our department from Jan. 1999 to Dec. 2012 were collected and analyzed in this study. The correlation between clinicopathological features and sensitivity of CA19-9/CEA was analyzed by chi-square test. The correlation of clinicopathological features and value of serum CA19-9/CEA was analyzed by t test and F test.</p><p><b>RESULTS</b>The average value of CA19-9 before surgery in the 126 patients was 595.3 U/ml. The values of CA19-9 in 91 patients were abnormal and the sensitivity of CA19-9 was 72.2%. The average value of CEA before surgery was 12.6 U/ml. The value of CEA in 26 patients were abnormal and the sensitivity of CEA was 20.6%. The values of combined detection of serum CA19-9 and CEA before surgery were abnormal in a total of 97 cases with a sensitivity of 77.0%. There was no significant correlation between clinicopathological features and sensitivity of CA19-9 (P > 0.05). The location of tumor was significantly correlated to the diagnostic sensitivity of CEA. The sensitivity of CEA to distal ECC was only 15.4%. The value of CA19-9 was relatively high in patients >60-year old or with neural invasion, while CEA was higher when tumor was located in the middle of bile duct (P < 0.05). There was no significant difference of serum CA19-9 before and after jaundice reduction (P > 0.05).</p><p><b>CONCLUSIONS</b>The diagnostic sensitivity of CA19-9 is not affected by gender, age, blood type, tumor location, degree of differentiation, tumor size, T stage, vascular tumor thrombus, lymph node metastasis, perineural invasion, and preoperative jaundice. However, the diagnostic sensitivity of CEA is affected by tumor location. The value of CA19-9 is correlated with tumor invasion and is relatively high in patients above 60 years old.</p>

Induced differentiation of human umbilical cord mesenchymal stem modified by cells Pdx1gene into islet beta-like cells in vitro / 生物医学工程学杂志

This study was to explore the induced differentiation of human mesenchymal stem cells (MSCs) modified by pancreatic and duodenal homeobox factor 1 (Pdx1) gene into insulin-producing cells in vitro. After recombined adenovirus vector with Pdx1 gene infected MSCs for 7 d, cells were induced by induction factors. The genes' expressions related to islet beta cells such as Pdx1, insulin, glucose transporter-2 (Glut2), were detected with RT-PCR, immunocytochemistry and Western blot. The levels of insulin and C peptide secretion were examined with chemiluminescence immunoassay. Insulin(+) cell rate was detected by flow cytometry. After infected by recombined adenovirus with Pdx1 and combined with induction factors, MSCs were aggregated and islet-like cell clusters formed. Dithizone staining of these cells was positive. The genes' expression related to islet beta cells, such as Pdx1, insulin, Glut2, could be detected. After induction, the islet-like cell clusters secreted insulin and C peptide. The levels of insulin and C peptide secretion increased with glucose stimulation. Insulin(+) cell rate was (11.61 +/- 4.83)%. It could be concluded that Pdx1 gene modified MSCs from human umbilical cord could be induced to differentiate into islet beta-like cells.

Enhanced effect of CD8++ T cells activated by tumor lysate -pulsed DCs on killing autologous tumor cells / 中国病理生理杂志

AIM:To evaluate the ability of dendritic cells (DCs) loaded with tumor lysate to initiate cell mediated immune responses by stimulating naive T cells, and the efficiency of activated T cells to kill autologous tumor cells in vitro. METHODS: The peripheral blood lymphocytes and monocytes were obtained from the advanced renal cell carcinoma patient by eonglutination method. The immature dendritic cells were generated in the presence of interleukin -4(IL-4) and granulocyte/macrophage colony-stimulating factor (GM-CSF) from monocytes of healthy individuals.These cells were pulsed with tumor lysate or not. Induction of tumor-specific cytotoxic T lymphocytes(CTLs) response by mature dendritic cells (mDCs) was evaluated by the CD95(Fas) expression assay through FCM and the cytotoxic assay a gninst autolognns human tumor cells. RESULTS: Human immature dendritic cells and T cells obtained from healthy donors were stimulated with tumor- pulsed dendritic cells. The immature dendritic cells were applied to the cytotoxicity assay a gainst target autologons tumor cells. The CD95 (Fas) expression, IFN-γ, and TNF -α secreted by the CTLs in tumor lysate-plused DC group were higher than those of other groups. The capacity of the CTLs to kill autolognns tumor cells was significantly different(P<0. 05). Antigen-specific DCs vaccine can induce T cells activation and proliferation, thus we can obtain higher proportion of tumor specific cytotoxic T cells(CTLs), and enhance the CTLs to secret IFN-γ and TNF-α. CONCLUSION: Our results indicate that monocyte-derived human dendritic cells pulsed with tumor lysate could in duce the specific antitumor effect against autologons tumors. This in vitro model offers a new and simple approach to the development of DC + CTL - based immunotherapy.

Dengue 2 virus induces molecular alteration in the coagulant and fibrinolytic pathways of human umbilical vein endothelial cells / 第三军医大学学报

Objective To study the changes of tissue plasminogen activator (t-PA), plasminogen activator inhibitor 1 (PAI-1), and soluble thrombomodulin (sTM) induced by dengue 2 virus (D 2V) infection of vascular endothelial cells. Methods The effects of D 2V infection on the production of t-PA, PAI-1, and sTM of human umbilical cord vein cells were studied. Results D 2V infection significantly induced the secretion of sTM and t-PA but showed no such effects on PAI-1 of human endothelial cells. Antibody against IL-6 inhibited D 2V-induced t-PA production of endothelial cells. A close correlation between serum levels of IL-6 and t-PA was found in dengue hemorrhagic fever (DHF) but not in dengue fever (DF) patients. Conclusion IL-6 can regulate D 2V-induced t-PA production of endothelial cells, suggesting that endothelial cells can be the target for D 2V infection and that D 2V-induced t-PA, TM, and IL-6 production of endothelial cells may contribute to the pathogenic development of dengue haemorrhagic fever/dengue shock syndrome (DHF/DSS).

Generation of insulin-producing cells from PDX1 and NKX6.1 gene-modified bone marrow mesenchymal stem cells / 中国病理生理杂志

AIM:To differentiate bone marrow mesenchymal stem cells(BMSCs) into functional insulin-producing cells to produce sufficient pancreatic islet cells for transplantation.METHODS:Recombinant adenovirus vectors carrying PDX1 and NKX6.1 genes were constructed and the bone marrow mesenchymal stem cells were infected by the recombinant adenovirus combined with several cytokines for differentiation.The expressions of PDX1,NKX6.1 and insulin and C-peptide in the differentiated bone marrow mesenchymal stem cells were detected by RT-PCR and Western blotting.After the differentiated bone marrow mesenchymal stem cells were transplanted into subrenal capsule of diabetic mice,cell morphology of the grafts as well as their secretion of insulin and C-peptide were detected.Besides,regulating capacities of grafts on the blood glucose level of the diabetic mice were also detected.RESULTS:BMSCs induced by recombinant adenovirus(pAdxsi-CMV-PDX1/CMV-NKX6.1) and several cytokines showed positive dithizone staining and the expressions of ?-cell related molecule such as insulin and glucose transporter 2 were detected by RT-PCR,which showed a sustaining and stable expression.The similar results were showed by Western blotting,immunohistochemical staining and indirect immunofluorescence.The insulin secretions in the cells stimulated with glucose at concentrations of 5.5 and 25 mmol/L in the experimental group were(1 240.4?109.3) mU/L and(3 539.8?245.1) mU/L,respectively,and were significantly higher than those in control group.Moreover,transplantation of the cells to STZ mice in treatment group made serum glucose recover to normal level.CONCLUSION:PDX1 and NKX6.1 gene-modified bone marrow mesenchymal stem cells differentiate into insulin-producing cells in vitro.When these cells transplanted into STZ induced diabetic mice,their serum glucose could return to the normal level and they could live well.Thus this is a promising method for diabetes treatment.

Significance of plasma soluable thrombomodulin in type 2 diabetic patients / 中华内分泌代谢杂志

Proteins in coagulative and fibrinolytic systems were measured in 50 type 2 diabetic patients and 50 normal controls. Proteins related with fibrinolytic system in the diabetic patients were significantly higher than those in normal controls. The concentration of soluble thrombomodulin (sTM) was negatively correlated with the activity of protein C and positively correlated with plasmin-? 2 -antiplasmin complex in type 2 diabetic patients, suggesting that the increase of sTM is associated with hypercoagulability and enhanced fibrinolysis.

Combination of Pdx1 and NeuroD1 results in Nkx6.1 and GLUT2 expressions in L02 cells / 中国病理生理杂志

AIM: To construct a recombinant plasmid vector containing human pancreatic duodenal homeobox1(Pdx1) and neurogenic differentiation 1(NeuroD1) genes,and to detect its effective expression in eukaryocytes and the ability to induce differentiation of hepatocytes.METHODS: Using human embryo pancreas mRNA as template,Pdx1 and NeuroD1 genes were amplified by RT-PCR and cloned into the two different multiple cloning sites(MCSA and MCSB) of plasmid pIRES.The recombinant plasmid pI/Pdx1/NeuroD1 was transfected into L02 cells.The expression of Pdx1 and NeuroD1 in transfected cells was detected by immunocytochemistry,IFA,RT-PCR and Western blotting,respectively.RESULTS: The length and sequence of cloned segments were correct.Pdx1 and NeuroD1 were expressed in eukaryocytes.Furthermore,the hepatic cells were induced to express glucose transporter 2(GLUT2) and eukaryocyte transcription factor Nkx6.1,which were functionally correlated to ? cells.CONCLUSION: pI/Pdx1/NeuroD1 plasmid is successfully constructed and expressed in human eukaryocytes,with which the cells express the eukaryocyte transcription factor and GLUT2,indicating the transfected cells functionally correlates to ? cells.The results suggest that Pdx1 and NeuroD1 genes can induce the differentiation the cells from hepatic cells to pancreatic endocrine secretion cells.

Enhanced effect of CD8~+ T cells activated by tumor lysate-pulsed DCs on killing autologous tumor cells / 中国病理生理杂志

AIM:To evaluate the ability of dendritic cells (DCs) loaded with tumor lysate to initiate cell-mediated immune responses by stimulating naive T cells, and the efficiency of activated T cells to kill autologous tumor cells in vitro. METHODS: The peripheral blood lymphocytes and monocytes were obtained from the advanced renal cell carcinoma patient by conglutination method. The immature dendritic cells were generated in the presence of interleukin-4(IL-4) and granulocyte/macrophage colony-stimulating factor(GM-CSF) from monocytes of healthy individuals. These cells were pulsed with tumor lysate or not. Induction of tumor-specific cytotoxic T lymphocytes (CTLs) response by mature dendritic cells (mDCs) was evaluated by the CD95(Fas) expression assay through FCM and the cytotoxic assay against autologous human tumor cells. RESULTS: Human immature dendritic cells and T cells obtained from healthy donors were stimulated with tumor-pulsed dendritic cells. The immature dendritic cells were applied to the cytotoxicity assay against target autologous tumor cells. The CD95(Fas) expression, IFN-? and TNF-? secreted by the CTLs in tumor lysate-plused DC group were higher than those of other groups. The capacity of the CTLs to kill autologous tumor cells was significantly different(P

Research advance in molecular characteristics of human stanniocalcin 1 / 中国病理生理杂志

Stanniocalcin(STC) is a glycoprotein hormone that was firstly found in bony fish.The related human proteins,STC1and STC2,are expressed in a wide variety of tissues.STC1 is involved in calcium and phosphate homeostasis,and plays important roles in carcinogenesis.This article reviews the data currently available regarding the human STC1,and discusses the roles they may play in normal physiology and in cancers.

LPS increases plasminogen activator inhibitor type-1 expression in human endothelial cells / 中国病理生理杂志

AIM: The aim of this study is to elucidate the effects of lipopolysaccharide (LPS) on tissue plasminogen activator(tPA) and plasminogen activator inhibitor type-1(PAI-1) expression and secretion in endothelial cells. METHODS: Cultured human umbilical vein endothelial cells (HUVECs) were induced by LPS for different times. Cell viability was then determined by cell counting kit-8. tPA and PAI-1 activities in the media were assayed by fibrin overlay and reverse fibrin autograph, respectively. Cytoplasmic RNA was prepared using the Trizol method and was assayed for PAI-I and tPA mRNA levels by reverse transcript-polymerase chain reaction(RT-PCR). RESULTS: LPS(10 mg/L) did not produce cell toxicity according to LDH determination in culture media. PAI-1 activity in LPS group was high (P0.05). CONCLUSION: LPS (10 mg/L) did not show signs of cell toxicity, but promoted the expression of PAI-1 mRNA and induced an increase activity of PAI-1. However, LPS (10 mg/L) did not change tPA mRNA expression. The time-dependent increase in PAI-1 mRNA expression and activity shifts the local balance toword increased anti-fibrinolytic capacity, which can amplify the extent of acute thrombosis after plaque rupture. This is one of the possible reasons that cause thrombus,blood coagulation and disseminated intravascular coagulation (DIC) during septicemia.