Objective To observe the morphological change of retina and its affection on visual function after optic nerve crush in adult rat. Methods The models of optic nerve crush in rats were made. The change of retinas in different time (1,3 ,5 ,7,9,14,28,56,84d respectively) after injury were observed by light microscope, and the flash visual evoked potential (F-VEP) was recorded in normal and injured rats. Result The number of retinal ganglion cells (RGCs) was significantly reduced in the partial lesion of optic nerve crush compared with normal optic nerves without injury. The initial loss of RGCs was accelerated during 3-7days after nerve crush and then declined gradually by 14 days, and no change was observed after 28 days. The wave of F-VEP became lower and wider in one day after the crush, and the latency and amplitude of F-VEP declined gradually from one day to 14 days, and recovery response was observed by the time of 4 weeks after injury. Conclusion The secondary degeneration of RGCs following optic nerve crush is the important morphological foundation of fall of visual function, which have relativity with the regular decline of visual function.

Objective To study the changes of transforming growth factor-β (TGF-β),connective tissue growth factor (CTGF) and collagen 1 (COL1) in newborn rat's lung tissue and the expressions of 4EBP1 (eukaryotic promoter) and P7OS6K (mammalian target of rapamyein pathway downstream target protein) after rapamycin and hyperoxia intervention,and to study the influence of mammalian target of rapamyein (mTOR) pathway on hyperoxic lung injury and the possible intervention methods.Method A total of 48 21-day-old neonatal rats were assigned into 8 groups (n =6),including air control group,3 d group (3 days after hyperoxic exposure),7 d group (7 days after hyperoxic exposure),14 d group (14 days after hyperoxic exposure),air + RAPA group (air + rapamycin),3 d + RAPA group (3 days after hyperoxic exposure + rapamycin),7 d + RAPA group (7 days after hyperoxic exposure + rapamycin) and 14 d + RAPA group (14 days after hyperoxic exposure + rapamycin).In the hyperoxic group,newborn rats were exposed to 90％ oxygen for 3,7,14 days respectively.The rats in the hyperoxia + rapamycin intervention groups received intraperitoneal injection of rapamycin and inhaled high concentrations of oxygen for 3,7,14 days respectively.Air ± rapamycin group received intraperitoneal injection of rapamycin for 3 days.To study the pathological changes of lung tissures after hyperoxia and rapamycin intervention,we used ELISA to detect the changes of TGF-β,CTGF and COL1 and Western blot to detect the variations of mTORC1,P70S6K and 4EBP1 expression.Result TGF-β,CTGF,COL1 levels at 3 days,7 days and 14 days after hyperoxic exposure (TGF-[β:33.7±2.8 vs.58.6 ±3.1 vs.98.8 ±1.5 ng/mg,CTGF:50.1 ±1.8 vs.68.7 ± 2.2 vs.94.4 ±2.5 ng/mg,COL1:471.9 ±5.7 vs.529.7 ±7.0 vs.556.4 ±8.5 ng/mg) were significantly higher than the air control group (TGF-β:25.5 ± 1.9 ng/mg,CTGF:41.7 ± 1.4 ng/mg,COL1:414.4 ± 8.9 ng/mg) (P ＜ 0.01).While the levels in rapamycin intervention group were significantly lower than all the hyperoxia + rapamycin intervention groups (P ＜ 0.01).The lung tissue pathological grades in 3 d + RAPA group and 7 d + RAPAgroup were significantly lower than those in the 3 d group and 7 d group (3.5 ± 0.8 vs.6.3 ± 2.3 and 9.7 ± 2.0 vs.14.0 ± 2.4) (P ＜ 0.01).The mTORC1,P70S6K,4EBP1 expressions in 3 d + RAPA group were lower than 3 d group (mTORC1:0.26 ± 0.04 vs.0.29±0.08,P70S6K:0.29±0.01 vs.0.31 ±0.08,4EBP1:0.31 ±0.06 vs.0.33 ±0.06) (P＜0.05),while the expressions in 7 d + RAPA and 14 d + RAPA groups were significantly lower than 3 d + RAPAgroup (P ＜0.01).Conclusion mTOR signal pathway may be involved in the repairing process of hyperoxic-induced lung fibrosis.Rapamycin can reduce the levels of TGF-β,CTGF and COL1 and inhibit the expressions of mTOR pathway downstream target protein P70S6K and 4EBP1,thus reduce lung injury atearly stage.

BACKGROUND: Skin defect is commonly repaired by autologous skin graft, but in which, it is required healthy skin provider and it probably results in scarring deformity to various extents. The successful construction and clinical application of tissue-engineered skin (TE skin) mark the major breakthrough in treatment of skin defect.OBJECTIVE: To analyze the relationship between operation method and healing rate, through repair of skin defect with TE skin, to provide experimental evidence on clinical application of TE skin.DESIGN: Randomized controlled observation was designed.SETTING: Department of Oral and Maxillofacial Surgery, Teaching-Research Room of Histology and Pathology and Experimental Center of Tissue Engineering, School of Stomatology, Fourth Military Medical University of Chinese PLAMATERIALS: The experiment was performed in Experimental Center of Tissue Engineering, School of Stomatology, Fourth Military Medical University, in which 6 healthy York pigs were employed, of clean grade,aged varied from 2.5 to 3 months. 3 groups were randomized, named TE whole-layer group, TE dermal and auto-epidermal group and auto-graft group, 2 pigs in each group. 8 wounds were prepared in each pig, round in shape and 50 mm in diameter, 16 wounds in each group, totally 48wounds.METHODS: ①Preparation of TE whole layer and TE true skin. ② In TE whole-layer group: The whole layer of skin was cut off from fat layeralong the drawn line. When bleeding stopped thoroughly and the wound was covered with wet physiological saline gauze, TE whole-layer skin was collected and windowing was done on the skin for drainage. Physiological saline was used to rinsed away the culture solution on the surface of TE skin, and then, the cuticular layer was upward-covered the wound, avoiding gas vacuole between cuticular layer and wound. Single-layer oleic gauze, physiological saline gauze, aseptic dry gauze and elastic sponge cushion were covered successively, about 3-5 mm in thickness each layer. After routine dressing, elastic bandage was wrapped with compression terminally. ③ TE dermal and auto-epidermal group: The whole- layer skin was cut off with same method. Thin split-thickness skin (TIS) 0.1-0.2 mm was collected with drum dermanuring machine and soaked in physiological saline. The same method was used to collect the managed TE true skin and cover it on the wound, covering immediately on autoTTS. The rest management was same as TE whole-layer group. ④ Autograft group: The whole-layer skin was cut off and the fat tissue was removed, afterwards, it was re-grafted on the auto-wound, covered with various layers of dressing and bandaged with compression. ⑤ The survival case was determined if it was discovered no infection, necrosis and scaling of grafted skin, less than 3 mm in diameter when the wound was opened for changing fresh dressing each time, otherwise, the failed case was recorded. The survival rate in each group was analyzed statistically in 4 weeks after operation.MAIN OUTCOME MEASURES: Survival situation of grafted skin in 4weeks after operation in each group.RESULTS: In 4 weeks after operation, the survival rate of grafted skin was 75% in TE whole-layer group was 87% in TE dermal and auto-epidermal group and was 94% in auto-graft group. The results were similar basically in comparison among 3 groups (x2=-2.34, P ＞ 0.05).CONCLUSION: The effect of TE skin graft on repair of skin defect is near to that of auto-epidermal graft, testifying that the repair of skin defect with TE skin is feasible.

Objective To evaluate the effect of greater trochanteric transfer for the treatment of coxa brevis in children and adolescents.Methods From August 2005 to January 2011,twenty patients (22 hips)with coxa brevis underwent greater trochanteric transfer.Among them 18 patients (20 hips) were available for evaluation,including 4 boys and 14 girls,with an average age of 11.4 years (range,7.5-15.0 years) at operation.Five cases (6 hips) were caused by Perthes disease,and 13 cases (14 hips) were caused by developmental dysplasia of hip.Four patients (4 hips) had undergone greater trochanteric epiphyseodesis ever before.All patients were fixed with tension screw after the deformity was corrected during the operation.After operation,the human plaster spica was used for 6 weeks in all patients.Results All patients were followed up for 14 to 79 months (average,31 months).At the last follow-up,fatigue or pain in the hips disappeared or improved in 13 patients.Sixteen patients had limping and positive Trendelenburg sign preoperatively,at the last follow-up 9 patients got improvement.Twelve patients (13 hips) had limitation of abduction of the hip,the average range of abduction was 25.38°±1.20°,which was improved to 45.38°±1.05° at the last follow-up.The average articulotrochanteric distance and ratio of the distance from the greater trochanter tip to femoral head center and the radius of the femoral head at the last follow-up was (17.47+3.14)mm and 2.10±0.21,respectively,there were statistical differences compared with those before operation [(-2.89±4.62) mm and 1.59±0.22,respectiovely].The average leg-length discrepancy at the last follow-up was (0.78t±0.26) cm,which had on statistical differences compared with that [(0.83 ±0.33) cm]before operation.Conclusion Greater trochanteric transfer for the treatment of coxa brevis in children and adolescents could improve the clinical symptom,recover the normal anatomy of the proximal femoral,restore the hip biomechanics environment,but could uot improve the leg-length discrepancy.

Objective Deleting the cysteine-rich region (22-37 amino acids)of HIV-1 HXB2 Tat protein(whole length is 101 amino acids) to improve its stability and expression level in E.coli and to analyze the immunogenicity of Tat protein without the cystein-rich region [Tat(△C)protein]. Methods Tat DNA deleted the cysteine-rich region (64-111 nucleotides), named as Tat(△C)DNA, was obtained in vitro by PCR and cloned into pET-32a vector. pET-32a-Tat(△C)plasmid and the pET-32a-Tat plasmid were established and transformed into E.coli BL21(DE3) strains respectively to express and purify the protein. Three rabbits were vaccinated with pET-32a-Tat(△C)protein, then testify the reactivity of sera from rabbits by ELISA and Western blot. Results The dense of the purified pET-32a-Tat(△C)protein was 7.12 mg/ml,which was greatly more than pET-32a-Tat protein(1.50 mg/ml). Dimer of pET-32a-Tat protein can be observed just after the protein purification and stored at 25℃ and 4℃ for 7 days, but dimer of pET-32a-Tat(△C)protein was not formed at the same condition. Experimental rabbits were immunized with pET-32a-Tat(△C)protein and produced high titre of anti-pET-32a-Tat(△C)serum(1∶320 000), the antibody can react specifically with Tat(△C)protein, Tat protein (1-101 AA)and synthetic Tat(1-86 AA) protein. Deletion mutation of the cysteine-rich region of Tat protein was first performed in the study. Conclusion The expression level in E.coli and the stability of Tat protein deleted the cysteine-rich region can be increased greatly, and the protein remains good immunogenicity. The results may provide a novel antigen for further development of HIV-1 Tat vaccine.

OBJECTIVE To discuss the related factors of toxigenic Helicobacter pylori(Hp) infection among adult population in Futian District,Shenzhen.METHODS A total of 1164 adults who had health examination in community centers were investigated through questionnaire and detected Hp antibodies.RESULTS The Hp and toxigenic Hp infection rates among them were 45.5% and 14.2%.Among them 46.3% and 14.8% were in males,and 44.7% and 13.4% in females.People less than 60 years old had higher incidence of Hp and toxigenic Hp,but decrease in people rnore than 60 years old.Hp and toxigenic Hp infection rate in married people were higher than those of unmarried.In workers,farmers and doctors were higher than staff and other occupations.The incidence were higher in drinking raw water than those drinking boiling water.in taking antibiotics before serologic examination were lower than no taking antibiotics;in peptic ulcer disease were higher than in no peptic ulcer disease;in per capita housing area over 6 m2 were higher than less 6 m2.CONCLUSIONS The infection rates of Hp and toxigenic Hp among our adult population are low,and positively correlated with age but gender.The social environment such as living condition,economic income,culture level and residential density maybe influence the infection of Hp.The route of transmission of Hp is unknown yet,water perhaps is an important dissemination medium.

Objective To explore the clinical efficacy of knee osteoarthritis treated by abdominal acupuncture combined with sodium hyaluronate injected in intraarticular.Methods 120 patients with knee osteoarthritis were randomly divided into observation group and control group,60 patients in each group.The observation group was treated by abdominal acupuncture combined with sodium hyaluronate injected in intraarticular.The control group was treated with intraarticular injection of sodium hyaluronate alone.Results After a follow up of 6 months, the total effective rate of the treatment group was 95.00%,which was higher than 78.33% of the control group, the difference was statistically significant (x2=7.21,P<0.01).Conclusion Abdominal acupuncture combined with intraarticular injection of sodium hyaluronate is a good way to treat middle and old age patients with knee osteoarthritis.

Objective:To investigate the influence of chemotherapy on the phenotype of secreted protein, acidic and rich in cysteine (SPARC) in gastric cancer (GC). Methods:Immunohistochemistry was used to analyze SPARC expression in 132 GC patients. Among these patients, 54 with preoperative chemotherapy and 78 without preoperative chemotherapy were selected to analyze the effect of chemotherapy on SPARC phenotype by comparing the postoperative specimens of the two cohorts. Results:SPARC expression was higher in GC lesions than in the desmoplastic stroma surrounding the tumor cells and noncancerous tissues. High SPARC expression was related to invasion depth, lymph node metastasis, and TNM staging. SPARC expression was lower in patients with preoperative chemotherapy than in controls ( P<0.05). Gross type, histology, depth of invasion, lymph node metastasis, TNM staging, and SPARC phenotype correlated with the overall survival of the patients with preoperative chemotherapy. Further multivariate analysis suggested that lymph node metastasis, histology, and SPARC phenotype after chemotherapy were independent prognostic indicators of GC. Conclusion:SPARC expression was associated with invasion depth, lymph node metastasis, TNM staging and GC prognosis. Preoperative chemotherapy may change the phenotype of SPARC in GC patients.

BACKGROUND:There are few studies concerning estrogen receptorβgene, and its mechanism of regulating the bone metabolism is stil unclear now. OBJECTIVE:To analyze the effect of estrogen receptorβ(ERβ) silencing on the expressions of transforming growth factorβ1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2) in human osteoblasts METHODS:There were three groups:blank control group (hFOB 1.19 uninfected with any retrovirus);negative control group (containing invalid interference fragment ERβ-shRNA-nc);optimal RNAi group (ERβ-shRNA-3). ERβ-shRNA retroviral vectors in the optimal RNAi group were used to transfect human osteoblasts fol owed by resistance screening and cel expansion. MTT assay was used to detect the proliferative activity of ERβ-silenced osteoblasts. Then under estrogen intervention, the stable inhibition rate of ERβwas determined using western blot assay, and the expressions of TGF-β1 and BMP-2 in human osteoblasts after ERβsilencing were detected by RT-PCR technology and western blot assay. RESULTS AND CONCLUSION:Human osteoblasts that were stably transfected by ERβ-shRNA-3 retroviral vector was selected successful y, and ERβsilencing had no significant influence on the cel proliferation (P>0.05). Under the interference of estrogen, the silencing efficiency of ERβprotein was (93.11±0.57)%(P<0.05), and after ERβsilencing, the expressions of TGF-β1 and BMP-2 were increased by (26.65±3.81)%and (16.62±1.71)%at mRNA level, and increased by (23.79±3.76)%and (18.08±3.20)%at protein level (both P<0.05). In conclusion, ERβmay play an important role in bone metabolism by regulating the expressions of TGF-β1 and BMP-2.

Objective To investigate the feasibility of self‐control materials serum thyroid hormone detected in the clinical labo‐ratory internal quality control in applications .Methods The serum thyroid hormone test results in patients with high‐value ,the ser‐um was mixed with a certain proportion of the healthy population ,dubbed serum control materials ,after aliquots -20 ℃ .Every o‐riginal supporting quality control and simultaneous detection of free thyroxine (FT4) ,three free triiodothyronine (FT3) ,thyroid stimulating hormone (TSH) ,thyroglobulin (TG) ,thyroid‐microglobulin antibody (TMA) ,thyroglobulin antibodies (TGA) and other thyroid hormone six items detected before the appearance of control samples was observed continuously detected six months to observe stability ;used EXCELL do J‐L maps ,statistical mean of each project within six months ,CV and uncontrolled number of observed values in the internal quality control applications .Results Within 6 months after the product was frozen homemade quali‐ty control serum soluble appearance clear and transparent ,no precipitation and turbidity ;1 ,3 ,6 ,10 ,15 ,30 ,45 ,60 ,90 ,120 day of the first magnitude and no significant changes in thyroid hormone ;compared with the original quality control materials ,low CV values . In the six months internal quality control applications ,the cumulative results of each project without 1 ,3 ,6‐month and six months significantly different ;project and six months mean cumulative comparison between the mean drift or change in trend did not ap‐pear ;items and cumulative CV were in control of the allowable range (CV <1/3 times within total variation );6 months of internal quality control work ,runaway alarm :13s ,2 times ,22s ,1 times ,R4s ,1 times .Conclusion Serum thyroid hormone homemade frozen control samples have better stability ,and can replace its import control materials ;internal quality control in their daily activities , through EXCELL test results for J‐L diagram precision can find instability timely corrective treatment to ensure that thyroid func‐tion test results are reliable ,worthy of clinical application .