Trends in clinical islet transplantation / 中华器官移植杂志

Islet transplantation has developed rapidly over the last 20 years and is becoming one of the ideal clinical treatment options for type 1 diabetes mellitus (T1DM). There is growing clinical evidence that islet transplantation has significant efficacy in insulin independence or reduction, preventing hypoglycemic episodes and preventing diabetic complications.However, clinical islet transplantation still faces challenges, such as a shortage of donor resources, difficulties in early implantation and survival of islet grafts, and immune rejection.In the future, donor pancreatic donation and its effective utilization should be promoted, and clinical exploration of xenogeneic islet transplantation and stem cell-derived islet transplantation should be encouraged to effectively solve the problem of insufficient islet source.At the same time, through continuous research and development of new materials and technologies, optimize the location of islet transplantation to improve the implantation and survival of islet grafts, and gradually eliminate the need for immunosuppression.In addition, we should actively promote the development and application of post-islet transplantation graft monitoring tools to further ensure the long-term survival of post-islet transplantation grafts, so that more diabetic patients can benefit from it.

JNK inhibitor SP600125 inhibits interleukin-1β induced pyroptosis in βTC-6 cells / 中华内分泌代谢杂志

Objective:To investigate whether interleukin(IL)-1β is involved in pyroptosis which leads to mouse islet β cell line βTC-6 cell damage, and to explore the role of JNK inhibitor SP600125 in inhibiting IL-1β induced βTC-6 cell pyroptosis.Methods:βTC-6 cell line and mouse islets were incubated with IL-1β for 48 h or intervened with both JNK inhibitor SP600125 and IL-1R antagonist IL-1Ra, then GSDMD expression and β cell pyroptosis morphology were detected by immunofluorescence staining of GSDMD and DAPI. The expression levels of Gsdmd, IL-1β and IL-18 mRNAs were detected by real time fluorescence PCR, and apoptosis was examined by Annexin-V/7-AAD staining combined with flow cytometry.Results:βTC-6 cell pyroptotic body was significantly increased in the IL-1β treated group compared with the control group, and the expressions of pyroptosis related genes Gsdmd, IL-1β, and IL-18 mRNA were significantly higher( P<0.05), and apoptosis was increased, suggesting that IL-1β effectively induced the βTC-6 cell pyroptosis, IL-1Ra prevented IL-1β induced βTC-6 cell pyroptosis. In the presence of JNK inhibitor SP600125, IL-1β treatment failed to induce the expressions of Gsdmd and IL-18 mRNA, markers of pyroptosis, and reduced the rate of apoptosis, indicating that SP600125 suppressed IL-1β induced βTC-6 cell pyroptosis. Conclusion:Pyroptosis is one of the mechanisms of βTC-6 cell impairment caused by IL-1β, and SP600125, a JNK inhibitor, can block the IL-1β induced pyroptosis pathway and has a potential role in inhibiting βTC-6 cell pyroptosis.

Influence of donor and pancreatic characteristics on the success rate of islet isolation / 中华内分泌代谢杂志

Objective:To explore the selection criteria of the donor for islet transplantation of Chinese people by analyzing the correlation between pancreas characteristics and success rate of islets isolation.Methods:Data from 113 cases of human islet isolation were collected. According to the result of islet isolation, the donors were divided into two groups, the success group(IEQ≥250 000, purification≥30%, and viability≥80%), and the failure group(IEQ<250 000, or purification<30%, or viability<80%). The modified Ricordi method was used to digest pancreas tissue, and the continuous density gradient method was performed to purify islets. The islets were identified by staining with the Dithizone(DTZ), the islets were analyzed for cell viability and purity.Results:The donor age in success group was significantly younger than failure group in the range of age eligible for this study( t=2.479, P=0.015). Pearson correlation showed that donor age was positively corelated with islet yield( r=-0.214, P=0.047). There was more fat on the pancreas surface in the successful islet isolation group( z=-2.007, P=0.045). The digestibility( t=2.133, P=0.035) and recovery rate( t=5.912, P=0.001) were elevated in success group. Conclusion:The pancreases from younger donors could obtain the higher-yielding islet, the pancreas with more surface fat or with higher weight was associated with islet isolation success in the scope covered by the inclusion criteria of this study.

ALDH1A3 + INS + cells in non-diabetic subjects and risk for type 2 diabetes: Analysis of cross-sectional data / 中华内分泌代谢杂志

Objective:To investigate the correlations of β cell dedifferentiation in non-diabetic subjects with risk factors for type 2 diabetes mellitus(T2DM).Methods:Immunofluorescence staining with insulin and β cell dedifferentiated marker ALDH1A3 was used to evaluate the β cell dedifferentiation levels in 38 non-diabetic and 23 T2DM. Correlation analyses were performed between β cell dedifferentiation levels and available clinical parameters including age, body mass index, HbA 1C level, triglycerides, and cholesterol levels in non-diabetic subjects. Results:β cell dedifferentiation level defined by the positive expression of ALDH1A3 in β cells(ALDH1A3 + INS + cell proportion) was significantly elevated in T2DM subjects( P<0.001). In PreD subjects, ALDH1A3 + INS + cells proportion were decreased( P=0.050) and negatively correlated with HbA 1C( r=-0.44, P=0.006), but not with age and body mass index. The analysis of correlation with lipidemic parameters showed that ALDH1A3 + INS + cells proportion was positively correlated with plasma total cholesterol level( r=0.39, P=0.045), but not plasma total triglyceride. Conclusion:ALDH1A3 + INS + cells were found to be decreased in prediabetes, suggesting that there may be enhanced β-cell identity in prediabetes to compensate for insulin secretion requirements; ALDH1A3 + INS + cells were elevated in people with high plasma total cholesterol levels, suggesting that total cholesterol may be one of the factors that induce β-cell dedifferentiation.

Combined Laparoscopic spleen-preserving distal pancreatectomy and autologous islet transplantation: report of one case and literature review / 中华器官移植杂志

Objective:To Eveluate the safty and clinical efficacy of combined laparoscopic spleen-preserving distal pancreatectomy and autologous islet transplantation in the treatment of solid pseudopapillary neoplasm.Methods:A 22 years old solid pseudopapillary neoplasm female patient who underwent distal pancreatectomy and an autologous islet transplantation at Tianjin First Central Hospital, clinical date for 6 months follow up was collected and analyzed.Results:The patient was well recovered after surgery, and during the post-operative follow up, the fasting blood glucose was 5.72 mmol/L, HbA1c was 6.1%, remained insulin independent, the liver function was kept well.Conclusions:Combined Laparoscopic spleen-preserving distal pancreatectomy and autologous islet transplantation can effectively prevent diabetes after distal pancreatectomy.

Adipose Tissue-Derived Stem Cells from Type 2 Diabetics Reveal Conservative Alterations in Multidimensional Characteristics

Background and Objectives@#Adipose tissue-derived mesenchymal stem cells (ASCs) are recognized as an advantaged source for the prevention and treatment of diverse diseases including type 2 diabetes mellitus (T2DM). However, alterations in characteristics of ASCs from the aforementioned T2DM patients are still obscure, which also hinder the rigorous and systematic illumination of progression and pathogenesis. @*Methods@#and Results: In this study, we originally isolated peripancreatic adipose tissue-derived mesenchymal stem cells from both human type 2 diabetic and non-diabetic donors (T2DM-ASCs, ND-ASCs) with the parental consent, respectively. We noticed that T2DM-ASCs exhibited indistinguishable immunophenotype, cell vitality, chondrogenic differentiation and stemness as ND-ASCs. Simultaneously, there’s merely alterations in migration and immunoregulatory capacities in T2DM-ASCs. However, differing from ND-ASCs, T2DM-ASCs exhibited deficiency in adipogenic and osteogenic differentiation, and in particular, the delayed cell cycle and different cytokine expression spectrum. @*Conclusions@#The conservative alterations of T2DM-ASCs in multifaceted characteristics indicated the possibility of autologous application of ASCs for cell-based T2DM treatment in the future.

Gene variant analysis of a patient with multiple carboxylase deficiency / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a patient featuring multiple carboxylase deficiency (MCD).@*METHODS@#PCR and Sanger sequencing were used to detect variant in the coding region of BT and HLCS genes in the patient. Suspected variants were verified in her parents and 80 unrelated healthy controls by a PCR-restriction fragment length polymorphism (PCR-RFLP) method.@*RESULTS@#The patient was found to carry compound heterozygous variants of the HLCS gene, namely c.286delG (p.Val96Leufs*162) and c.1648G>A (p.Val550Met). The c.286delG (p.Val96Leufs*162) was verified to be novel variant based on the result of PCR-RFLP analysis. No variant was found in the coding regions of BT gene in the patient.@*CONCLUSION@#The compound c.286delG (p.Val96Leufs*162) and c.1648G>A (p.Val550Met) variants probably underlie the MCD disorder in this patient. Above results have enriched the variant spectrum of MCA.

Effect factors of liver enzymes elevation afterintraportal islet transplantation / 中华器官移植杂志

Objective@#To investigate the effect factors of liver enzymes elevation by monitoring the liver function changes before and after intraportal islet transplantation.@*Methods@#16 diabetic patients who received intraportal islet transplantation in our hospital were analyzed. The levels of aspartic aminotransferase (AST), alanine aminotransferase (ALT)and total bilirubin (TBil)were monitored after islet transplantation.@*Results@#Among those 16 diabetic patients who received intraportal islet transplantation, 11 patients showed an increased AST and 8 patients showed an increased ALT, among which a 2.5-fold increase in AST was observed in 4 patients and over 1.5-fold elevation of ALT was observed in 3 patients. The level of TBil were in the normal range before and after transplantation in all patients. Transplanted tissue volume of islet was the main factor for significantly increased AST (P<0.05) in this study. It is also shown that the change in portal pressure is related to the AST elevation after islet transplantation.@*Conclusions@#The amount of transplanted islet tissue volume is related to the liver enzymes elevation after intraportal islet transplantation. Therefore, the improvement of the purity of islet to reduce the amount of transplanted tissue could be benefit to prevent the liver injury after islet transplantation. Meanwhile, the purified islets should be injected as slowly as possible to maintain a stable portal pressure.

Effect factors of liver enzymes elevation afterintraportal islet transplantation / 中华器官移植杂志

Objective To investigate the effect factors of liver enzymes elevation by monitoring the liver function changes before and after intraportal islet transplantation .Methods 16 diabetic patients who received intraportal islet transplantation in our hospital were analyzed .The levels of aspartic aminotransferase (AST ) ,alanine aminotransferase (ALT )and total bilirubin (TBil)were monitored after islet transplantation .Results Among those 16 diabetic patients who received intraportal islet transplantation ,11 patients showed an increased AST and 8 patients showed an increased ALT ,among which a 2 .5-fold increase in AST was observed in 4 patients and over 1 .5-fold elevation of ALT was observed in 3 patients .The level of TBil were in the normal range before and after transplantation in all patients . Transplanted tissue volume of islet was the main factor for significantly increased AST (P< 0 .05) in this study .It is also shown that the change in portal pressure is related to the AST elevation after islet transplantation .Conclusions The amount of transplanted islet tissue volume is related to the liver enzymes elevation after intraportal islet transplantation .Therefore ,the improvement of the purity of islet to reduce the amount of transplanted tissue could be benefit to prevent the liver injury after islet transplantation .Meanwhile ,the purified islets should be injected as slowly as possible to maintain a stable portal pressure .

Comparison of different collagenases in islet isolation from Chinese organ donors / 中华器官移植杂志

Objective To retrospectively compare the efficacy of Serva NB1 collagenase with Vitacyte GOLD collagenase on islet isolation of pancreas.Methods All the human pancreata were obtained from Chinese organ donors.In GMP laboratory,the pancreata were trimmed and distended with Serva NB1 collagenase (Serva NB1,n =12) or Vitacyte GOLD collagenase (Vitacyte GOLD,n =5) and digested according to a modified Ricordi semi-automatic protocol,and the digestion duration was recorded.The digested islets were then collected and washed,followed by the continuous density purification in a Cobe 2991 cell separator.The islet yield,purity,viability and glucose-stimulated insulin release (GSI) were determined each time after purification.Quantity and quality of isolated islets were determined by digestion efficacy.Results The digestion duration in Vitacyte GOLD collagenase group was significantly shorter than in Serva NB1 collagenase group to achieve the same digestion endpoint (P＜ 0.05).The islets yields of different sizes were variable between the two groups.The Vitacyte GOLD collagenase digestion produced more islets with a diameter range of 50-100 μm than the ServaNB1 collagenase digestion (P＜0.05),but the latter yielded more islets with a diameter range of 251-300 μm and 301-350μm (P＜0.05).There was no significant difference in total islets yields,viability,and GSI between two collagenase digestions (P＞0.05).Conclusion Both Vitacyte GOLD collagenase and Serva NB1 collagenase can be used for the clinical islet isolation in China.

Role of autophagy in hydrogen-induced inhibition of apoptosis in hippocampal neurons in a rat model of orthotopic liver transplantation / 中华麻醉学杂志

Objective To evaluate the role of autophagy in hydrogen-induced inhibition of apoptosis in hippocampal neurons in a rat model of orthotopic liver transplantation (OLT).Methods Fifty-six pathogen-free healthy adult male Sprague-Dawley rats,aged 8-10 weeks,weighing 220-250 g,were used in the study.Thirty-two rats were selected and assigned into 4 groups (n =8 each) using a random number table:sham operation group (group S),OLT group,hydrogen-rich saline group (group HS) and chloroquine group (group CQ).The other 24 rats severed as the donors.In group S,laparotomy was performed,and the related blood vessels were isolated.The model of OLT was established in OLT,HS and CQ groups.In group OLT,normal saline 6 ml/kg was slowly injected via the inferior vena cava at 5 min before anhepatic phase.In group HS,hydrogen-rich saline 6 ml/kg was slowly injected via the inferior vena cava at 5 min before anhepatic phase.In group CQ,autophagy inhibitor chloroquine 60 mg/kg was injected intraperitoneally at 1 h before establishment of the model,and the other treatments were similar to those previously described in group HS.At 6 h of reperfusion,the rats were sacrificed and hippocampi were isolated for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity,for pathological examination (with light microscope),and for detection of cell apoptosis (by TUNEL staining) and expression of autophagy-and apoptosis-related proteins caspase-3,cytochrome c (Cyt c),microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ),Beclin-1 and p53 in hippocampal tissues (by Western blot analysis).Apoptosis index (AI) was calculated.Results Compared with group S,the MDA content and AI were significantly increased,the SOD activity was decreased,and the expression of caspase-3,Cyt c,LC3 Ⅱ,Beclin-1 and p53 was up-regulated in OLT,HS and CQ groups (P＜0.05).Compared with group OLT,the MDA content and AI were significantly decreased,the SOD activity was increased,the expression of caspase-3 and Cyt c was down-regulated,and the expression of LC3 Ⅱ,Beclin-1 and p53 was up-regulated in group HS (P＜0.05).Compared with group HS,the MDA content and AI were significantly increased,the SOD activity was decreased,and the expression of caspase-3 and Cyt c was up-regulated,and the expression of LC3 Ⅱ,Beclin-1 and p53 was down-regulated in group CQ (P＜0.05).Conclusion The mechanism by which hydrogen inhibits apoptosis in hippocampal neurons is related to promotion of autophagy in a rat model of OLT.

Advances of target therapy in hormone-receptor-positive breast cancer / 中国肿瘤临床

Hormone therapy is a major treatment option for hormone-receptor-positive (HR+) breast cancer. Resistance to hormone therapy is the major reason for disease recurrence and progression. Recent studies have identified several resistance mechanisms that lead to the estrogen-independent growth of HR+breast cancer and were exploited to develop novel target drugs, including cyclin-de-pendent kinases 4 and 6 (CDK4/6) inhibitor, mammalian target of rapamycin (mTOR) inhibitor, epidermal growth factor receptor family inhibitor, vascular endothelial growth factor inhibitor, histone deacetylase inhibitor, fibroblast growth factor receptor inhibitor, insulin-like growth factor receptor inhibitor, and checkpoint inhibitor. These inhibitors are being developed to block resistance pathways and improve the efficacy of hormonal therapy. Among these drugs, the mTOR inhibitor everolimus and the CDK4/6 inhibitor palbociclib are currently approved in the United States to treat metastatic HR+breast cancer. In this study, we summarize the frontier advances in the combination of hormone and target therapies.

One case of concurrent hemorrhage after pancreatic islet transplantation by percutaneous transhepatic portal approach / 中华危重病急救医学

The percutaneous transhepatic portal approach is the most commonly used technique for islet transplantation, largely owing to its safety and minimally invasive characteristic. Bleeding complications after islet transplantation are rare. A case of type 1 diabetes mellitus (T1DM) was treated in Tianjin First Center Hospital, who had a massive intra-abdominal hemorrhage after percutaneous transhepatic portal vein catheterization for islet transplantation. Through the review of the overall development of the case, we aim to improve the awareness of the complications of islet transplantation, to reduce the incidence of complications after percutaneous transhepatic portal vein transplantation, and to provide experience.

Role of Nrf2/HO-1 signaling pathway in reduction of acute kidney injury following orthotopic liver transplantation by hydrogen-rich saline in rats / 中华麻醉学杂志

Objective To investigate the role of nuclear factor erythroid 2-related factor 2 (Nrf2) / heme oxygenase-1 (HO-1) signaling pathway in reduction of acute kidney injury following orthotopic liver transplantation (OLT) by hydrogen-rich saline in rats.Methods Thirty-two healthy adult male SpragueDawley rats,weighing 220-250 g,were randomly assigned into 4 groups (n =8 each) using a random number table:sham operation group (S group),OLT group,hydrogen-rich saline group (HS group),and all-trans retinoic acid (ATRA) group.Laparotomy was performed,and the related blood vessels were isolated in group S.The model of orthotopic autologous liver transplantation was established in OLT,HS and ATRA groups.Normal saline and hydrogen-rich saline 6 ml/kg were injected through the inferior vena cava at 5 min before the portal vein was clamped in OLT and HS groups,respectively.In group ATRA,Nrf2 inhibitor ATRA 7 mg/kg was injected intraperitoneally once a day for 2 consecutive days,the model of orthotopic autologous liver transplantation was established at 16 h after the last injection of ATRA,and the other treatments were similar to those previously described in group HS.At 6 h of reperfusion,blood samples were collected for determination of serum blood urea nitrogen (BUN),creatinine (Cr),interleukin10 (1L-10) and tumor necrosis factor-alpha (TNF-α) concentrations.After blood sampling,the lungs were removed for determination of malondialdehyde (MDA) content,superoxide dismutase (SOD) activity,expression of HO-1,Bcl-2 and Bax mRNA (by using real-time reverse transcriptase polymerase chain reaction),and HO-1 protein expression in lung tissues (by Western blot) and for microscopic examination.The damage to the renal tubules was scored.Results Compared with group S,the serum BUN,Cr and TNF-α concentrations were significantly increased,the serum IL-10 concentrations were decreased,the MDA content and renal tubular damage score were increased,the SOD activity was decreased,and the expression of HO-1 protein and mRNA,and Bcl-2 and Bax mRNA was up-regulated in group OLT (P＜ 0.05).Compared with group OLT,the serum BUN,Cr and TNF-α concentrations were significantly decreased,the serum IL-10 concentrations were increased,the MDA content and renal tubular damage score were decreased,the SOD activity was increased,the expression of HO-1 protein and mRNA and Bcl-2 mR-NA was up-regulated,and the expression of Bax mRNA was down-regulated in group HS (P＜0.05).Compared with group HS,the serum BUN,Cr and TNF-α concentrations were significantly increased,the serum IL-10 concentrations were decreased,the MDA content and renal tubular damage score were increased,the SOD activity was decreased,the expression of HO-1 protein and mRNA and Bcl-2 mRNA was down-regulated,and the expression of Bax mRNA was up-regulated in group ATRA (P＜0.05).Conclusion The mechanism by which hydrogen-rich saline reduces acute kidney injury following OLT is probably associated with activation of Nrf2/HO-1 signaling pathway in rats.

Clinical characteristics and prognosis of invasive lobular carcinoma of breast: a report 125 cases / 肿瘤研究与临床

Objective To analyze clinicopathologic characteristics and prognostic factors of the patients with invasive lobular carcinoma of breast. Methods Clinical data of 125 patients with invasive lobular carcinoma of breast treated at Cancer Center of Sun Yat-sen University from Jan. 1990 to Dec. 2008, were analyzed. The clinical characteristics, recurrence and survival of the patients were summarized. Results Median age of 125 patients was 45 years old (range, 27 to 76 years old). The patients with large tumor mass (≥ 3cm), positive local lymph node, more than Ⅱ stage and positive hormone receptor at diagnosis were 77 cases(61.6 %), 64 cases(51.2 %), 101 cases(80.8 %) and 112 cases(89.6 %), respectively. The median time of follow-up was 58 months (range, 11-222 months). Of the 125 patients, 32 had local recurrence and metastasis, and 18 died. The 5-year disease-free and overall survival rates were 82.2 % and 87.3 %, respectively. Multivariate COX regression analysis showed that whether endocrine therapy or not was only a prognostic factor of patients with invasive lobular carcinoma of breast. Conclusion There is no difference in media age of patients with invasive lobular carcinoma of breast at diagnosis from other pathologic type of breast cancer. These patients are usually with larger tumor masses, more lymph node metastasis and a higher proportion of positive hormone receptor. The prognosis of patients is not affected by clinicopathologic parameters.

Chromosomal and Subcellular Localization and Expression of Cell Cycle-related Regulator DCT1 / 中国医科大学学报

Objective To investigate the chromosomal and subcellular localization of DOC-1R terminal 1(DCT1),and detect its expression in human tissues.Methods Chromosome localization of DCT1 was detected by radiation hybrid.pEGFP-DCT1 was constructed,and HeLa cells were transfected with the plasmid.The subcellular localization of DCT1 protein was observed by fluorescence microscope.Real-time PCR was performed for the determination of DCT1 expression level in 16 kinds of human tissues.Results DCT1 was demonstrated to localize in 5q31,and its encoding protein was detected on the nuclear membrane.Additionally,DCT1 was proved to express universally in all the 16 kinds of human tissues and it was expressed at the highest level in spleen.Conclusion DCT1 might be a regulator in cell cycle,and ubiquitously express in human tissues.

Characteristics of micro-spatial structure of artifical nerve bridging substance for repairing nervous segmental defect / 中国组织工程研究

BACKGROUND: There is no any mature and complete system to repair nerve tissue after segmental injury due to particularity of nerve tissue regeneration. Using tissue-engineering technique to solve this problem is difficult and challenged.OBJECTIVE: To research an artificial nerve bridging substance in application of repairing nerve injury on clinic and obverse its microcosmic spatial structure at the same time.DESIGN: Open experiment.SETTING: Orthopaedic Research Institute, Xijing Hospital of the Fourth Military Medical University of Chinese PLA.MATERTALS: The experiment was carried out in the Orthopaedic Research Institute, Xijing Hospital of the Fourth Military Medical University of Chinese PLA from November 2001 to January 2003. CollagenⅠ, collagen Ⅳ and gelatin were provided by Sigma-aldrich Company, USA.METHODS: Establishment of bridging substance: Collagen Ⅰ (65.53%) and collagen Ⅳ (32.77%) were added into 0.05 mol/L acetic acid solution, respectively; and then, two suspensions were mixed together and stirred at 4℃. The suspension of collagen and heparin sulfate was pumped, held still and poured into silica gel tube with bore of 3 mm.Continuously, two ends of tube were enclosed and suffered from cold drench and mold at five various speeds. General observation was done. Composites were intercepted cross section, vertical section and 45° section for observation by optic microscope at different speeds of 10×10-5 m/s, 5×10-5 m/s, 2.5×10-5 m/s, 1.0×10-5 m/s and 1.0×10-6 m/s; meanwhile, interior structure of microtubule was observed with scanning electron microscope. Interior diameter of microtubule was calculated as the following formula: actual aperture = (magnification × area of microtubule) ÷ (length of guage × perimeter of aperture).MAIN OUTCOME MEASURES: ① General observation; ② observation with microscope; ③ observation with scanning electron microscope and interior diameter of microtubule.RESULTS: ① General observation: Models were regular cylinders and had symmetrical features. Flexility was wonderful,texture was well-proportioned, and elasticity was strong. ② Results of optic microscope: Structure of external surface was completely blocking without any poricidal dehiscence. Surface was smooth and had good continuity. ③ Results of electron microscope: External surface shaped like terrace tile; asides, interior structure of microtubule was average and its courser was coincident and parallel to each other. Microtubules with longitudinal courser were independence on each other. The structure was blocking and did not connect to each other. This was as the same as courser of nerve fibrous bands of organis. Cross section of interior microtubule was general round and regular, and the diameter was average. Interior microtubule had groat continuity without breaks or transection. Trabeculae of microtubule also had groat continuity,and its surface was smooth. Interior diameter of microtubule ranged from 197.3 μm to 258.8 μm.CONCLUSION: Nerve bridging substance is made of collagen and gelatin which are characterized by groat compatibility and degradation after mixture, lysis and cold drench, It has singly longitudinal microtubule and microstructure of high imitation by normal nerves, so nerve bridging substance can be used in basic researches and repair nerve injury on clinic.

Changes of substance P and calcitonin gene-related peptide in rat spinal dorsal horn during feeling function recovery after late stage peripheral nerve injury / 中国组织工程研究

BACKGROUND: Whether the injured peripheral nerve in a late stage has repairing value still remains a problem. If irreversible changes happen in substance P and calcitonin gene-related peptide, the feeling function will lose even after repairing.OBJECTIVE: To quantitatively study the changes of substance P(SP) and calcitonin gene-related peptide(CGRP) in the spinal dorsal horn 24 weeks after peripheral nerve injury.DESIGN: A self-controlled quantitative experiment.SETTING: Institute of Orthopaedics of the Fourth Military Medical University of Chinese PLA.MATERIALS: This experiment was performed in the Institute of Orthopaedics of the Fourth Military Medical University of Chinese PLA from October 2002 to May 2003. Totally 55 SD rats were divided into 11 groups according to time points(1, 2, 3, 4, 6, 8, 10, 12, 16, 20 and 24 weeks after sciatic nerve transection).INTERVENTIONS: Sciatic nerve injury model was set up by transecting one side of sciatic nerve and ligating the proximal stump of sciatic nerve; the other side was set as the control side. Computer-assisted image analysis was used to measure the immunologic reaction areas of substance P and CGRP.MAIN OUTCOME MEASURES: Changes of the distribution of the positive fibres of SP and CGRP in rat spinal dorsal horn in each group.RESULTS: Fifty-five rats entered the result analysis. The distributions of SP immunoreactivity in the spinal dorsal horn following sciatic nerve injury showed a significant reduction during 2-6 weeks, followed by a slow rate of increase,and reached almost complete restoration at 16 weeks after deafferentation. No obvious advanced changes happened at 20 and 24 weeks. The ratios for ipsilateral and cotralateral sides of positive fibre and distribution area injury in spinal dorsal horn CGRP were 1.14 at week 1, 1.13 at week 6, and 0. 29 at week 24. The ratios were similar at each time point( P ＞ 0. 05).CONCLUSION: At the late stage of peripheral nerve injury, neurons in the spinal dorsal horn and dorsal root ganglion still remain their functions to synthesize and secrete SP and CGRP. Spinal dorsal horn remains at a balanced status and still has the neurologic basis to recover the sensory function.

sHLA-G1 inhibiting xenotransplantation rejection through degrading the releasing of porforin and granzyme by NK cells / 医学研究生学报

Objective: To study the effect of sHLA-G1 inhibiting xenotransplantation rejection through degrading the releasing of porforin and granzyme by NK92. Methods: The recombinant expression vector pcDNA3-sHLAG1 was transfected into the lymphoblastoid cell line LCL721.221 by the nucleofection methods.The expression of sHLA-G1 in the transfected LCL721.221 was detected by RT-PCR and DOT-ELISA technique.NK cell line(NK92) was used as NK effect cells and the porcine aortic endothelial cells line(PED) as targets.The inhibitive effects of the releasing of porforin and granzyme by NK92 was analyzed by the ?-hexosaminidase release assay.And the cytotoxicity of NK92 was analyzed by MTT methods. Results:sHLA-G1 conferred a significant degrading the releasing of porforin and granzyme by NK92,and protection PED against NK92 medicated lysis,and the rate of NK92 cell cytotoxicity was reduced to 25.5% in contrast to 71.2% in the control group(P