BACKGROUND: Collagen type Ⅰ possesses low antigenicity and provides special functions of scaffold structure for cell migration, proliferation and secretion.OBJECTIVE: To observe the morphological change after combination of transformed human embryonic tendon cells and artificial materials with collagen type Ⅰ, so as to provide the experimental basis for constructing tissue-engineered muscle tendon in vitro.DESIGN: Randomized and controlled observation.SETTING: Department of Orthopaedics, West China Hospital, Sichuan University.MATERIALS: This experiment was carried out at the Department of Transplantation Immunology, West China University of Medical Sciences from July to September 1998. Human hair, carbon fiber and polyglycolic acid were combined with collagen and then set corresponding groups respectively. Simply hair, carbon fiber and polyglycolic acid control groups were set respectively. Artificial materials: Healthy adult hair (30 cm in length, 80 g in weight), carbon fiber knitted strip (Lanzhou carbon factory )and polyglyolic acid fasciculi (Johnson , American) were used in this experiment.METHODS: ①In the group of human hair combined with collagen,nine healthy adult hairs were chosen . Three hairs as one bundle were knitted into tendon with 0.8 cm in length and 0.4 cm in diameter and a knot was tied respectively at the two ends. ②In the carbon fiber combined with collagen group, the purchased carbon fiber knitted strip was separated and made into artificial materials with the same diameter and length as those of human hair tendon. Two ends of the artificial materials were packed with 5/0 silk thread. ③In the polyglycolic acid combined with collagen group, three polyglycolic acid fasciculi were weaved into bundle with the same diameter and length as those of human tendon,and two ends were packed with 5/0 silk thread. ④The process of knitting scaffold materials was the same in control groups as in the corresponding experimental groups. ⑤ Five pieces were prepared in each scaffold material. After sterilized by 60Co, the scaffold materials were put in collagen type Ⅰ solution for 0.5 hour, then taken out and dried at roomtemperature for use in the experimental groups. ⑥transformed human embryonic tendon cell, density for 3×106/mm3, was used for combined culture in each group, and cell number and morphology were observed at different time points after culture under an inverted microscope and scanning electron microscope.MAIN OUTCOME MEASURES: ① Observation of cell growth and adherence under an inverted microscope ② Observation of cell morphology and material adherence under scanning electron microscope RESULTS: ① Observation of cell growth and adherenceunder an inverted microscope :In each experimental group, at two hours after culture, cells gathered around the materials, and some cells adhered to materials presented round; At three days after culture, the number of cells adhered to materials began to increase, morphology changed from round to oval, and the co-existence of round and oval cells was seen;While in each control group, at two and three hours after culture, few cells adhered to the three materials. On day five after culture, in each experimental group, cell morphology turned into fusiform, the number of cells increased further and a great many fusiform cells grew among carbon fiber materials; while in the control group, most cells presented round and ellipse, and the number of cells was less as compared with the corresponding experimental group. ② Cell morphology and material adherence under scanning electron microscope: On day 5 after in vitro culture, most tendon cells on the surface of materials presented spindle-shape in each experimental group. Apophysis stretched out from the tendon cells, grew along longitudinal axon of the materials. Silk and piece-like substance appeared among the apophysis and weaved each other. While there were fewer cells adhered in control group than in corresponding experimental group, cells also presented fusiform or round, but no silk or piece-like substance was found, also no apophysis appeared.CONCLUSION: Human hair, carbon fiber and polyglycolic acid combined with collagen type Ⅰ will facilitate cell adherence. Cells increase around the materials and co-existence of round and oval cells could be seen, and cells turned into fusiform, suggesting that exogenous collagen facilitate the cells to adhere onto the materials and proliferate.

Objective To investigate the effects of short-term continuous subcutaneous insulin infusion (CSII) in newly diag-nosed type 2 diabetes mellitus(T2DM ) and to evaluate the treatment regimen conversion after CSII therapy .Methods 72 patients with newly diagnosed T2DM were treated with CSII for 2 weeks .Then they were randomly divided into two groups :the basal insu-lin group(glargine) and the oral anti-diabetic drug(OAD) group .Both groups were followed up for 2 years .Blood glucose ,insulin and HbA1c were measured before and after CSII and during the 2-year follow-up .Results CSII significantly controlled the glucose levels ,reduced the TG ,TC ,LDL levels and the homeostasis model assessment of insulin resistance ( HOMA-IR) and increased the homeostasis model assessment of insulin secretion (HOMA-IS)(all P< 0 .05) .During 1-year follow-up ,HbA1c in the glargine group and the OAD group was (6 .13 ± 0 .47)% and(6 .21 ± 0 .38)% respectively .During 2-year follow-up ,the HbA1c values in the two groups were (6 .91 ± 0 .57)% and(6 .43 ± 0 .62)% respectively .T HOMA-IR and HOAM-IS obtained the long tern improve-ment without significant body weight increase .Conclusion Short-term intensive insulin therapy can effectively control the blood glucose ,improve the sensitivity of insulin and the β-cell function ;after CSII ,adopting basic insulin and oral antidiabetic drugs can a-chieve the ideal glycemic control .

a greater impact,which could account for some pathophysiological changes caused by catch-up growth.

Objective To investigate the characteristics and risk factors of abdominal lymph node metastasis in thoracic esophageal squamous cell cancer.Methods The clinical data of 586 patients with thoracic esophageal cancer who underwent surgery via transabdominal and transthoracic approaches between June 2009 and June 2014 at the Sichuan Cancer Hospital were retrospectively analyzed.All the patients received resection of esophageal cancer and lymph node dissection, and the transabdominal right thoracic approach or cervico-thoracicabdominal triple incision was selected according to the condition of patients.No.18, 19, 20 lymph nodes were dissected seperately and No.16, 17 and lesser curvature lymph nodes were separated.All the specimens of lymph nodes were detected by regular pathological examination.Measurement data with normal distribution were presented as x ± s and count data were described as rate.Comparisons of rate between 2 specimens and among the multiple specimens were respectively analyzed using the chi-square test and partition of chi-squared.The multivariate analysis was done using the logistic regression.Results The number of lymph node dissected in 586 patients was 12 524 with an average number of 20 ± 11 per case, and the rate of lymph node metastasis was 55.63％ (326/586).The number of mediastinal lymph node dissected was 7 012 with an average number of 12 ± 5 per case, and a rate of mediastinal lymph node metastasis was 40.96％ (240/586).The number of abdominal lymph node dissected was 5 512 with an average number of 9 ± 8 per case, and a metastasis rate was 31.74％ (186/586).The abdominal lymph node metastasis rate of the upper, middle and lower thoracic esophageal cancer were 13.73％ (14/102), 31.51％ (92/292) and 41.67％ (80/192), respectively, showing a significant difference among the above 3 indexes (x2 =25.91, P ＜ 0.05).The lymph node metastasis rate in No.16, 17, 18,19, 20 and lesser curvature lymph nodes were 12.80％ (75/586), 16.89％ (99/586), 1.71％ (10/586),0.68％ (4/586), 1.71％ (10/586) and 2.05％ (12/586), respectively, with a significant difference among the above 6 indexes (x2 =287.95, P ＜ 0.05).The results of univariate analysis showed that the tumor location,surgical procedure, T stage, N stage, G stage, pathological stage and mediastinal lymph node metastasis were risk factors affecting abdominal lymph node metastasis of thoracic esophageal cancer (x2 =24.02, 23.97, 37.87,136.85, 38.79, 7.70, 154.27, P ＜ 0.05).The tumor in the lower thoracic portion, N3 stage and stage Ⅳ were independent risk factors affecting abdominal lymph node metastasis of thoracic esophageal cancer in the multivariate analysis (RR =5.80, 2.36, 2.76, 95％ confidence interval: 1.022-1.813, 1.317-3.950, 1.652-12.351, P ＜ 0.05).Conclusions Abdominal lymph node metastasis is common in thoracic esophageal cancer in which No.16 and 17 lymph nodes predominate, and it is easy to occur in patients with lower thoracic esophageal cancer, and advanced N stage and pathological type.

Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.

To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.

AIM: To study the neuroprotective roles of neuroglobin (NGB) over-expression in the SH-SY5Y cells transfected with pAPPswe.METHODS:The plasmid pEGFP-NGB was successfully constructed and transfected into the SH-SY5Y cells, which were pretreated with pAPPswe.MTT assay was applied to detect the effect of NGB over-expres-sion on the cell survival rates.JC-1 staining was used to detect the level of mitochondrial transmembrane potential.The cell apoptosis was analyzed by flow cytometry.The effects of NGB over-expression on the protein level of p-Akt, Akt and caspase-3/9 were determined by Western blotting.The generation of Aβ42 in the cells was measured by ELISA.RE-SULTS:The cell survival rate was remarkably increased after transfection with NGB compared with control group and emp-ty plasmid group (P<0.05).The over-expression of NGB significantly inhibited the decrease in mitochondrial membrane potential induced by pAPPswe.The over-expression of NGB inhibited the apoptosis of the cells.Furthermore, over-expres-sion of NGB not only inhibited the expression of caspase-3 and caspase-9, but also induced the production of p-Akt, which was prevented by LY294002, an inhibitor of PI3K/Akt.The generation of Aβ42 was inhibited in the cells with the over-ex-pression of NGB.CONCLUSION: Over-expression of NGB significantly inhibits the SH-SY5Y cell injuries induced by pAPPswe and inhibits the expression of caspase-3/9, which is tightly related with cell apoptosis.Furthermore, the neuro-protective roles of NGB may be via activating PI3K/Akt signaling pathway.

OBJECTIVETo study the related risk factors for surgical site infection following Pilon fracture surgery. METH ODS: The data of 561 patients with Pilon fractures treated with open reduction plate osteosynthesis at our institution's trauma centre were collected from January 2006 to December 2012. All the patients were divided into two groups: infection group and non-infection group. In the infection group, there were 23 males and 10 females, ranging in age from 21 to 69 years old, with an average of (45.50±4.40) years old. In the non-infection group, there were 296 males and 232 females, ranging in age from 16 to 76 years old, with an average of (43.50±7.19) years old. The possible risk factors such as age, gender, smoking, diabetes, alcohol abuse, open fractures, compartment syndrome and operative time were studied. The multivariate Logistic regression model was used to analyze the risk, factors.

RESULTSThe infection rate of surgical site after Pilon fracture surgery was 5.88%. There were significant statistical differences between infection group and non-infection group in operative time, open fractures and compartment syndrome. However, multivariate Logistic regression analysis revealed that only operative time was significantly associated with surgical site infection (P=0.005, OR=44.92).

CONCLUSIONOperation time is an independent predictor for post-operative surgical site infection of Pilon fracture treated with open reduction plate osteosynthesis. Though open fracture and compartment syndrome could increase the surgical site infection rate, they could not not be considered as independent predictors.

Adult ; Compartment Syndromes ; complications ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Operative Time ; Risk Factors ; Surgical Wound Infection ; etiology ; Tibial Fractures ; surgery

Objective To explore clinical characters of type 1 diabetes mellitus with different serum potassium levels in children and adolescent.Methods One hundred and seventy-five patients with type 1 diabetes mellitus were reviewed,they were divided into 3 groups according to the serum potassium level.The patients whose serum potassium