Objective The expression of transforming growth factor beta receptor Ⅰ (TGF?RⅠ) and transforming growth factor beta receptor Ⅱ (TGF?RⅡ) in the synovium and the articular cartilage of patients with osteoarthritis were studied,to explore the possible relationship between the transforming growth factor beta receptor and the pathogenesis and treatment of osteoarthritis.Method The distribution and positive levels of TGF?RⅠ,TGF?RⅡ and TGF? 1 in the synovium and articular cartilage from 26 patients with osteoarthritis and 3 patients with trauma were studied,using Immunohistochemial methods.Results Immunohistochemical staining of TGF?RⅠ showed positive in all synovial samples in patients with osteoarthritis.The positive staining of TGF?RⅠ was found in most synovial lining cells,endotheliocyte and the macrophage in subsynovial layer with osteoarthritis,especially macrophage like synoviocyte.The positive particles were distributed in the cytoplasma.The distribution and staining levels of TGF?RⅡ in synovium and articular cartilage of patients with osteoarthritis were similar to those of TGF?RⅠ.Positive stainings for TGF?RⅠ, TGF?RⅡ and TGF? 1 were found in over half of chondrocytes.Conclusion It is suggested that vehicles of signal trasmission of TGF? are rich in the synovial membrane and cartilage.There is an important effect of TGF? receptor on inhibiting inflammatory process and helping to repair local tissue.

To investigate the changes in the activated T-lymphocyte CD3/HLA-DR and CD3/CD(16+56) populations in peripheral blood of the patients with allogeneic hand transplantation, lymphocytes from peripheral blood of the patients at different time points were immunologically labeled with dual color fluoresecent monoclonal antibodies CD3/CD(16+56) and CD3/HLA-DR, mono-color fluoresecent monoclonal antibody CD25. CD25, CD3/CD(16+56), and CD3/HLA-DR were determined with flow cytometry (FCM). The levels of activated T-lymphocyte (CD25 +,CD3 +/HLA-DR + ), silent T-lymphocyte [CD3 +/CD(16+56) -,CD3 +/HLA-DA - ] decreased significantly during the first week after transplantation and then increased gradually to the pre-operafive level. Nature killer cells [CD3 -/CD(16+56) +] increased significantly at the first day after transplantation, then decreased sharply and maintained a lower level. The results suggest that immunosuppressive agents have significantly effects on lymphocyte subsets in allogenaic hand transplanted patients, and dynamic determination of HLA-DR, CD3 /CD(16+56) could be valuable in immunomonitoring after allogeneichand transplantation.

Objective To study the interaction between human microRNA ( miRNA) and 5′trailer regions of Ebola virus genome from the perspective of bioinformatics so as to facilitate prevention and treatment of Ebola virus .Method The miRNA target prediction software Pita and RNAhybrid were used to predict the human miRNAs which could bind the 5′trailer regions of Ebola virus genomes before the miRNAs were annotated by g:Profiler web server .Results and Conclusion There may be complex interactions between human miRNAs and the 5′trailer regions of Ebola virus .Previous reports about the interaction between host miRNA and 5′trailer region of virus genome suggest that the interaction between human miRNA and 5′trailer region of Ebola virus may have effect on replication of Ebola virus and human cells .This work may provide new ideas on prevention and treatment of Ebola virus .

Objective To investigate the clinical results of posterior cruciate ligament (PCL) reconstruction by double bundle-double tunnel Y-shape of the anterior tibialis tendon allograft. Methods From March 2001 to January 2008, 47 patients underwent PCL reconstruction were included. The allogeneic adult anterior tibialis tendon was prepared into the Y-shape double bundles with the length of 130 mm; A bundle was defined as A-side; B-side was two short bundle (B1, B2 bundle). A bundle was 70 mm in length with a diameter of 10-12 mm. B1 bundle (anterolateral bundle) was 55 mm long with a diameter of 6 mm; B2 bundle(posteromedial bundle) was about 50 mm with a diameter of 6 mm. The allograft ligament was installed through the antero-medial approach. Absorbable interface screws were fixed in the tibial tunnel firstly, and then in the femoral tundles. When being fixed, anterolateral bundle was in flexion of 90°, postero-medial bundle was in 30°. Assisted exercise with knee an angle-locked walking aid had continued for 8-10 weeks. Results The average operating time were 45 min. The average follow-up time was 49.5 months. Preoperative Lachmann was positive in all cases while Lachmann was negative in 39 cases, weakly positive in 5 cases, and positive in 4 cases postoperatively. Post-operative KT-1000 testing, Lysholm score and Tegner activity levels has improved significantly compare with the pre-operative ones. Conclusion The double folded bundles of adult anterior tibialis tendon has sufficient length and diameter for posterior cruciate ligament reconstruction with power tension. The methods of ligament passing through the tunnel has improved to ease the procedure.

BACKGROUND: There are a lot of immunologic studies about limb allotransplantation in animal experiment. But, it is only early investigation in clinic; its clinical immunologic study needs further accumulation.OBJECTIVE: To dynamically analyze the early immunologic state change in patients following hand allotransplantation.DESIGN: Controlled trial.SETTING: Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA; Department of Traumatic Orthopaedics and Department of Laboratory Medicine, Nanfang Hospital, First Military Medical University of Chinese PLA.PARTICIPANTS: Two patients who underwent unilateral hand allotransplantation in the Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA were enrolled, serving as experimental group. The observation was between September 1999 and March 2000. Twenty persons, including 12 male and 8 female, who homochronously received health examination, aged 20 to 45 years, were enrolled, serving as healthy control group. They all had no reactive immune and infectious diseases, and voluntarily participated in the trial.METHODS: Peripheral blood was collected from 2 patients who underwent hand allotransplantation once respectively at pre-operative 1 day and 3 days. Blood collecting was performed once per day at post-operative 1 week, three times per week at post-operative 2 to 4 weeks, twice per week at 5 to 8 weeks post-operation, once per week at 9 to 16 weeks post-operation, twice per month at 5 to 6 months post-operation. ① Peripheral blood T cell subgroups (CD3+,CD4+,CD8+T cells)were detected by flow cytometer,serum panel reactive antibody (PRA) by ELISA method, serum C-reactive protein by turbidimetric immanoassay (TIA), serum creatine kinase (CK) by enzyme dynamics method. ②Mixed lymphocyte reaction(MLR): mitomycin C-treated donor peripheral blood lymphocytes were used as stimulator, and proliferative reaction of peripheral blood lymphocyte of patients to donor transplanted antigen was detected with the incorporation of 3H-TDR method (Negative: There was no significant difference between the mean value of stimulation index and 1, conversely positive). Autogenic peripheral blood lymphocytes treated with the same way replaced donor stimulator, serving as control. Stimulation index of each specimen was calculated (Stimulation index=Experiment cmp/controlcpm), serving as control index. Peripheral blood T-cell subgroups (CD3+,CD4+,CD8+T cell), serum PRA, C-reactive protein and CK were detected in 20 persons in healthy control group;Twenty persons were randomly divided into 10 groups. Two persons in each group were used as donor and recipient mutually and performed MLR.MAIN OUTCOME MEASURES: ① Peripheral blood T cell subgrpups (CD3+,CD4+,CD8+T cell). ②PRA. ③ C-reactive protein. ④CK. ⑤MLR.RESULTS: ①CD3+,CD4+,CD8+T cell levels were obviously decreased within one week after operation. CD3+ and CD4+T cell levels both recovered to be the pre-operative levels, but CD8+ level exceeded pre-operative level significantly [CD3+: (66.43±4.56); CD4+: (30.55±3.94); CD8 +:(33.45 ±2.69)]. There was no significant difference between experiment group and control group. ②Serum PRA was 0 to 10%, there was no significant difference as compared with control group. ③ Serum C-reactive protein was 0 to 0.359 mg/L, there was no significant difference as compared with control group. ④ Serum CK was 25 to 170 mmol/L, and there was no significant difference as compared with control group. ⑤ MLR after transplantation was negative, and it turned into be positive 5 months later.They were all positive in control group.CONCLUSION: Short-term change and long-term redistribution of T cell subgroups are closely related to immunosuppressive agent, suggesting that immunosuppressive agent has obvious effect on T-cell subgroup following hand allotransplantation. Immuno-induction schedule make patients be in immune suppression state, which effectively avoid early rejection. But patients cannot bear specificity yet; they need the inhibition of immunosuppressive agents.

BACKGROUND: Apoptosis plays a key role in intestinal mucosal ischemia-reperfusion injury and recovery; meanwhile, effect of shenfu injection on apoptosis of intestinal epithelial cells during intestinal mucosal ischemia-reperfusion injury should be studied further.OBJECTIVE: To investigate the relationship between the apoptosis of intestinal epithelium and characteristics of intestinal mucosal ischemia-reperfusion injury and recovery.DESIGN: Randomized controlled animal experiment.SETTING: Department of General Surgery, Xianning Central Hospital;Department of General Surgery, Renmin Hospital of Wuhan University.MATERIALS: The experiment was carried out in the Central Laboratory,Xianning Central Hospital from March to August 2005. Fifty-four healthy male SD rats weighting 200-250 g were provided by Animal Center of Medical School, Wuhan University.METHODS: The rats were divided randomly into 3 experimental groups:control group (n=6), ischemia-reperfusion group (n=24) and shenfu treatment group (n=24). ① Pentobarbital sodium solution (40 mg/kg) was administrated into the intraperitoneal cavity to induce anaesthesia. Through a midline abdominal incision, the mesenteric blood vessel of a 15-cm segment of mid-intestine was occluded for 60-minute with an atraumatic vascular forceps. The control group underwent the same procedure except for unblocking the mesenteric blood vessel. At the end of 60 minutes ischemia period the forceps was removed to allow reperfusion, the abdominal cavity was closed. ShenFu injection (8 mL/kg ·h, 20 mL/kg ·d, produced from Yaan Three-Nine Pharmaceuticals Co, No: 030302) was injected 30 minutes before occlusion in SF treatment group, same quantity of 0.9% natrii chloride was injected in control group and ischemia-regeneration group at the same time, and oxygen was inbreathed during the operation and ischemia-regeneration. ② Experimental intestinal canals were sampled for the following analysis when all groups were respectively performed sham ischemia for 1 hour, intestinal ischemia for 1 hour and reperfusion for 1, 24and 72 hours. Sections were observed in light microscope. Histological mucosal damage in each sample was evaluated as followed scoring system: 0score, normal muscosal villi and gland; 1 score, slight lesion near the tip of the villi; 2 scores, slight lesion of subepithelial gland; 3 scores, development of subepithelial (Gruenhagen) spaces near the tip of the villi with capillary congestion; 4 scores, extension of the subepithelial space with moderate epithelial lifting from the lamina propria; 5 scores, a few denuded villous tips; 6 scores, massive denuded villi; 7 scores, denuded villi with exposed lamina propria and obvious gland lesion; 8 scores, disintegrateon of the lamina propria; 9 scores, haemorrhage and ulceration. ③ The Tunel method (TdT mediated biotin-dUTP nick and labeling; TdT-Frag EL DNA fragmentation detection kit) was used. Inbrief, this method allowed the identification of apoptosis nuclei in tissue samples through DNA fragment and labeling. Apoptosis Index (AI) was set as the average number of apoptosis cells in per 100 cells by observing ten high power fields of adjacent villi and crypts. ④ The mitotic phase of crypt epithelial nucleus within intestinal mucosa was observed in intestinal sections stained with haematoxylin and eosin. The number of cells with nucleus mitotic phase was counted in ten adjacent mucosal crypts, which was taken as the index of mitotic activity of intestinal mucosal epithelial cell.MAIN OUTCOME MEASURES: Intestinal mucosal histopathological changes, apoptosis of intestinal mucosal epithelial cell and mitotic activity of intestinal mucosal crypt.RESULTS: All 54 rats were involved in the final analysis. ①) Scores of histopathological changes were (0.65 ±0.35) points in 1-hour ischemia group, (3.87±0.86) points in 1-hour reperfusion group and (0.65±0.35)points in 24-hour reperfusion group; which were lower than those in ischemia-reperfusion group [(7.11±1.01), (8.05±1.34), (1.53±0.48) points; P＜ 0.05]. ② Indexes of apoptosis were 17.24±7.05 in 1-hour ischemia group, 24.20±9.87 in 1-hour reperfusion group, 11.49±4.71 in 24-hour reperfusion group and 6.02±2.16 in 72-hour reperfusion; which were lower than those in ischemia-reperfusion group (51.09±13.76, 54.89±15.58,23.54±9.64, 12.47±5.52; P ＜ 0.05). Activities of mitosis were 10.37±2.03and 11.72±2.07 in 1-hour ischemia group and 1-hour reperfusion group,respectively; which was higher than those in ischemia-reperfusion group(8.24±1.69, 9.95±1.93; P ＜ 0.05).CONCLUSION: Shenfu injection can significantly attenuate apoptosis of intestinal epithelium, increase crypt mitotic activity, and promote intestinal epithelium regeneration or repair.

Objective To study dynamic changes of T cell subsets and CD3＋HLA-DR＋T cells in peripheral blood of composite tissue transplantation-hand allograft recipients. Methods The levels of CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD3＋HLA-DR＋T cells and ratio of CD4/CD8 in peripheral blood of hand allograft recipients in different periods were examined by using flow cytometry.The recipients before transplantation were as the control groups.Results The first day after transplantation,the levels of CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD3＋HLA-DR＋T cells and ratio of CD4/CD8 all began to descend.The 3th to 5th day after transplantation,the levels were the lowest.The 8th day,the levels of CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD3＋HLA-DR＋T cells and ratio of CD4/CD8 were eventually rised and go to stable 15th day after transplantation.But the levels of CD3＋、CD3＋CD4＋and value of CD4/CD8 were still lower than those of the contols,and the levels of CD3＋CD8＋、CD3＋HLA-DR＋T cells were higher distinctly.Conclusion Dynamic changes of T cell subsets and active T cells in peripheral blood of composite tissue transplantation-hand allograft recipients were accordant with that of renal allograft recipients with stable period and with stable clinical state of the hand transplantation recipients.

Objective To develop a simple and quick method for detection of stress-induced 5′transfer RNA( tRNA) halves.Methods Total RNA purified from stress induced cells was polyadenylated by poly( A) polymerase, and then degen-erate DNA probes were used to hybridize with 3′tRNA-halves of intact tRNAs,while RNase H specifically degraded the 3′tRNA-halves strand in tRNA-DNA probes hybrids.Using the RNase H digestion total RNA as templates, complementary DNA( cDNA) was synthesized by oligo ( dT) n-anchored primers.The primer of 5′tRNA halves and anchored-primer were used to amplify 5′tRNA halves by PCR.Results The results showed that the method of poly ( A )-tailed-RNase H digestion-RT-PCR could be successfully used to detect stress-induced 5′tRNA halves.Conclusion A simple and quick method for detection of 5′tRNA halves has been established,which is a user-friendly tool for 5′tRNA halves detection and function research.

Objective To investigate the characteristics of click -auditory brainstem response (ABR) in nor-mal infants of 1 to 6 months old ,and to establish the normative values for latencies of Wave I ,III ,V and interpeak latencies of I- Ⅲ ,III-V and I-V for younger infants .Methods Click auditory brainstem responses were recorded from infants within 6 months :166 infants of 1 -months old(269 ears) ,141 2 -month old (226 ears) ,111 3 -months old(177 ears) ,58 4-months old(96 ears) ,78 5-months old(121 ears) and 45 6-months old(76 ears) .We compared the latencies of wave I ,III ,V and interpeak latencies of I - Ⅲ ,III-V ,I-V obtained from infants of differ-ent ages at different stimulus intensities .Results The average threshold of 1 to 6 months infants was 16 .18 ± 5 .35 dB nHL ,the average latency of Wave V was 9 .03 ± 0 .49 ms .The differences among the thresholds were statistical-ly insignificant(P>0 .05) .Wave I ,III and V were noticeable in all ears tested at 80 dB nHL .Wave I disappeared first as the stimulus intensity decreased ,and the latencies of Wave I ,III and V prolonged;on the contrary ,interpeak latencies of I -III ,III-V ,I-V shortened significantly .At the same stimulus intensity ,the latencies of Wave III , V and the interpeak latencies of I - Ⅲ ,III-V ,I-V shortened significantly except for Wave I .When comparing the differences among the testing parameters as a function of each month ,we found that there were statistically signifi-cant differences for the latencies of wave III ,V and the interpeak latencies of I -III ,III-V ,I-V before the 4 months old(P<0 .05) ,and there were no significant differences after 4 months old(P>0 .05) .Conclusion It is recommen-ded that 16 .18 ± 5 .35 dB nHL be used as the normative references for the evoked threshold of click auditory brain-stem responses for 1~6 month old infants .The development of central nervous system below the inferior calicles is fast before the 4 months old .

Objectives To assess the clinical value of ultrasonography (US) and bioelectrical impedance analysis (BIA) in analyzing abdominal fat contents of obese children and adolescents through comparison with MRI. A correlation with other obese related metabolic parameters was conducted. Methods Ninety 7-17-y-old obese children and adolescents (60 boys and 30 girls with mean age of 9.6 ± 2.9 y and mean BMI of 24.5 ± 4.5 kg/m2) were recruited. Metabolic parameters were measured, and insulin resistance was estimated according to homeostasis model assess-ment (HOMA-IR). On the same day abdomen subcutaneous fat thickness (SFTUS) was measured by US. Body fat mass (FMBIA) and abdominal visceral fat area (VFABIA) were analyzed by bioelectrical impedance analysis (BIA). After obtaining informed consent, abdominal MRI was performed in 20 subjects. Each section of umbilicus level was analyzed by image threshold value segmentation using SigmaScan Pro 5 and abdominal subcutaneous fat area (SFAMRI) and visceral fat area (VFAMRI) were calculated. Results (1) A strong positive association was found between SFTUS and SFAMRI (P< 0.05), VFABIA and VFAMRI (P < 0.01) respectively. (2) FMBIA and SFAMRI, VFAMRI, SFTUS also showed significant correlations (P < 0.05). (3) VFAMRI showed extremely significant positive correlations with TG, Insulin,C-peptide and HOMA-IR (P < 0.01 ) ; SFAMRI was also correlated positively with them (P < 0.05). (4) SFTUS was correlated positively with UA (uric acid), Insulin, 2HIns (insulin measured at 2 hours after meal), C-peptide,2HC-peptide (C-peptide measured at 2 hours after meal) and HOMA-IR (P < 0.01). (5) VFABIA was correlated significant positively with UA, insulin, TG, 2HIns and HOMA-IR. FMBIA showed positive correlation with UA, Insulin,2HIns, C-peptide, 2HC-peptide and HOMA-IR. Conclusions abdominal subcutaneous and visceral fat of obese children and adolescents evaluated by US and BIA are correlated well with those assessed by MRI, and also correlated well with TG, insulin, C-peptide and other metabolic biochemical parameters. Our data support the value of using cost effective, simple and convenient methods such as BIA and US to evaluate the obese and related metabolic risk of children and adolescents in clinical practice.