Objective To analyze the clinical features of limb salvage surgery with epiphyseal preservation in children and adolescents, and to evaluate the recurrence rate, metastasis, complications, and the joint functional results after tumor resection. Methods From December 1995 to January 2003, we operated on 33 patients aging from 8 to 16 years (average 12.2 years) with diagnosis of primary malignant bone tumor of the extremities with preserving the epiphysis in the limb salvage surgery. In this group, the tumor located in distal femur in 24 cases and in proximal tibia in 9 cases. There were 23 osteosarcomas, 6 Ewing sarcomas, 2 chondrosarcomas, and 2 invasive osteoblastomas. The tumors were staged clinically, 2 cases inⅠA, 2 cases inⅠB, 17 cases in ⅡA, and 12 cases inⅡB. According to the relationship between the tumor and epiphyseal plate, the metaphyseal tumors in children were classified into 3 types: 18 cases were categorized as type Ⅰ, 13 as type Ⅱ, and 2 as type Ⅲ. The patients received 2-4 cycles of preoperative adjuvant chemotherapy, and another 6 cycles after surgery. Serious bone defects after tumor resections were repaired with massive allograft bone transplantation, followed by internal fixation by intramedullary nails and cancellous screws. Results Among the 33 cases, 3 cases were lost to follow up. 29 cases had complete clinical data. Postoperative follow-up was 12-72 months (average 37.6 months). Recurrences were seen in 3 cases, with one local recurrence in type Ⅲ case one year after tumor resection, other 2 recurrences around the femoral vessels in typeⅡcases 15 and 30 months after the tumor resection respectively. The recurrence rate accounted for 10.3%. Amputation was performed for the cases with recurrences. But pulmonary metastasis developed and the patients died. In this cohort, 9 cases died. Five-year survival rate was 57.9%. 4 cases reported 5 complications (17.2%). No patient reported skin necrosis, hematoma, infection, rejection to the allograft, and nonunion at the end of the follow-up. According to the functional evaluation criteria of Enneking, excellent were 11 cases, good 13 cases, fair 3 cases, and poor 2 cases. The excellent or good rate was 82.8%. Conclusion The limb salvage surgery with preservation of epiphysis for malignant bone tumors in children and adolescents guarantees the patient's satisfactory postoperative limb functional results. Preoperative effective adjuvant chemotherapy and prevention of postoperative complications deserve great attention.

Child A.The concentration of CGRP varied positively with the diameter of portal vein(r=0.60,P

Objective To explore the diagnostic effect of serum level of cystatin C (CysC) on the renal function after neonatal asphyxia by detection of serum level of CysC, blood urea nitrogen (BUN) and serum creatinine (SCr) and calculation of glomerular filtration rate (GFR) in neonatal asphyxia. Methods The clinical data of 86 neonates with asphyxia (46 cases in mild asphyxia group,40 cases in severe asphyxia group) and 30 neonates without asphyxia (control group) were collected and the serum level of CysC, BUN and SCr were detected at 24 h to 72 h after birth. Results Serum levels of CysC, BUN and SCr were (1.97 ±0.33) mg/L, (4.97 ±2.15) mmol/L, (90.41 ±24.32) μmol/L in mild asphyxia group, (2.65 ±0.41) mg/L, (10.88 ± 3.31) mmol/L, (125.82 ± 45.44) μ mol/L in severe asphyxia group and (1.24 ± 0.35)mg/L, (4.25 ± 2.04) mmol/L, (58.41 ± 19.22) μmol/L in control group, respectively. The differences were significant among three groups and those values in mild and severe asphyxia groups were higher than those in control group. The sensitivity of CysC level to evaluate renal function in mild asphyxia group was better than BUN and SCr level (P＜ 0.05). In neonata] asphyxia, the serum level of CysC had negative correlation with GFR (P ＜ 0.01). Conclusions Serum level of CysC can be adopted to evaluate the renal function after neonatal asphyxia, which is better than BUN and SCr. With a higer level of CysC, the renal function injury may be worse.

Objective To explore the clinical characteristics of Pierre Robin syndrome and the experience of its diagnosis and therapy, and provide the accurate diagnosis and treatment of Pierre Robin syndrome .Methods Seventeen cases of Pierre Robin syn-drome from September 2008 to August 2013 were enrolled in this study .The clinical data were analyzed retrospectively to explore the clinical characteristics and treatment of Pierre Robin syndrome .Results Seventeen cases of Pierre Robin syndrome had typical clini-cal features, including micrognathia, cleft palate or high palatine arches, glossoptosis, and feeding difficulties.The babies were given corresponding treatment according to their clinical grading. Among those babies, three cases died , including one dying of severe pneu-monia in hospital, and the other two dying of severe pneumonia and malnutrition after giving up treatment .Among 14 survivors, 12 ba-bies were followed up , including 5 babies who achieved optimal growth and development when they were one year old , and the other 7 babies who had feeding difficulties in varying degrees after discharge , and lagged behind normal children in the growth and develop-ment.Conclusions Early diagnosis, accurate classification, and individualized treatment plan for Pierre Robin syndrome might im-prove the prognosis of children with this type of disease .

Objective To investigate the role of HMGB1 in the pathogenesis of organ injury of acute necrotizing pancreatitis (ANP).Methods Male ICR mice were randomly allocated into control group,ANP group and HMGB1 monoclonal antibody group.ANP model was induced by intraperitoneal injection of 20％ L-arginine.Mice in HMGB1 monoclonal antibody group were given intraperitoneal injection of 200 μg of HMGB1 monoclonal antibody immediately after the induction of the ANP model.All the mice were sacrificed at 12,24,and 48 h after ANP induction.Serum level of amylase and liver,renal function were determined,level of serum HMGB1 was measured by using enzyme-linked immunosorbent assay,and then the pathologic changes of pancreas and liver were routinely observed and scored.The HMGB1 mRNA levels in the liver and pancreas were studied by real time fluorescence quantitative PCR.Results The serum levels of HMGB1 at 12 h in control group,ANP group and HMGB 1 monoclonal antibody group were (9.09 ± 1.03),(25.04 ± 4.30),(16.84 ± 4.27) μg/L; and pathological scores of pancreatic tissue were (1.50 ± 0.55),(4.33 ± 0.52),(3.03 ± 0.32) points ; and HMGB1 mRNA expressions in pancreas were 0.48 ± 0.18,7.53 ± 2.71,3.26 ±2.33 ; HMGB1 mRNA expressions in liver were-1.23 ± 0.37,0.15 ± 0.65,－ 1.27 ± 0.72.The corresponding values in ANP group were significantly higher than those in control group (P ＜ 0.05).While the corresponding values in HMGB1 monoclonal antibody group were significantly lower than those in ANP group (P ＜0.05).There was a positive linear relationship between serum HMGB1 level and pancreatic pathological scores 24 h after ANP induction (r =0.768,P ＜ 0.05).In addition,the serum levels of AMY,AST,ALT,LDH,BUN,Cr showed a similar trend as that of serum level of HMGB1,and the serum level of HMGB1 was positively associated with serum levels of Cr,BUN and ALT (r =0.824,0.719,0.590,P＜0.05).Conclusions HMGB1 may be a key factor of inflammatory response and organ dysfunction of ANP in mice,and extrinsic supply of its monoclonal antibody may decrease the injuries of pancreas and other organs during ANP.

Objective To observe the clinical effect of modified Zhisou powder and Symbicort Turbuhaler simultaneously on patients with Cough Variant Asthma. Methods 120 patients with Cough Variant Asthma were randomly recurited into two groups. 60 patients in the treatment group were treated with modified Zhisou powder and Symbicort Turbuhaler; 60 patients in the control group were treated with Symbicort Turbuhaler. 4 weeks was a therapeutic course in both group. Results The markedly controlled rate (MCR) (clinical control+excenence)of the treatment group was 83.3%, obviously surpassed the control group (70.0%) (P＜0.05); There was obvious improvement of cough, expectoration, breath lessness and throaty pruritus after the therapy in both groups, but it was much better in the treatment group than the control group(P＜0.05). The pulmonary function was significantly improved after treatment in both groups(FEV1, FEV1% and PEF, P＜0.05). The improvement showed significant difference between the two groups(P＜0.05). There was obvious decrease of EOS, IL-5 and ECP in both groups. The decrease of ECP and IL-5 in the treatment group was greater than the control group(P＜0.01). Conclusion The therapy of modified Zhisou powder and Symbicort Turbuhaler has advantage over pure western therapy.

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. The expression of Sj14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

Objective To observe the serum levels of high mobility group box-1 protein (HMGB1)in patients with acute cholangitis (AC) and to investigate contributions of HMGB1 in AC.Methods Serum HMGB1 concentrations were determined by an enzyme-linked immunosorbent assay in 30 patients with AC of severe type (ACST) and 42 patients with mild acute cholangitis at the time of admission (within 72 h after the onset).A total of 50 healthy subjects were recruited as the control group.Fluorescent quantitative PCR (FQPCR) was used to detect the HMGB1 mRNA expression and the relationship between serum HMGB1 levels and clinical factors was analyzed.Results The serum HMGB1 levels in healthy control group,mild group and ACST group were (1.82 ± 0.64) μg/L,(10.46 ± 3.75) μg/L,(18.89 ± 6.86) μg/L,respectively.The mean value of serum HMGB1 level in mild group was significantly higher than that in control group,while significantly lower than that in ACST group (P ＜ 0.05).Compared to the control group,the HMGB1 mRNA level in patients of AC increased significantly and the level of ACST group was higher than that of mild group.The serum HMGB1 levels of patients with positive bile or/and blood cultures were higher than that of negative.After emergency endoscopic nasal biliary drainage,the serum HMGB1 levels of patients significantly decreased compared to preoperational (P ＜ 0.05).The HMGB1 levels were significantly positively correlated with white cell counts,C-reactive protein (CRP),total serum bilirubin,direct bilirubin and alkaline phosphatase (ALP).By logistic regression analysis,serum HMGB1 levels had correlation with severity of disease.Conclusion Serum HMGB1 levels significantly increased in patients with AC and the serum concentrations of ACST group were higher than those of mild group.Serum HMGB1 level has a correlation with sepsis.ENBD could lower its serum levels.Serum HMGB1 has predictive value to severity of disease.

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sjl4 or pVAC-Sj26 only to get two gene fragments including Sjl4 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES,resulting in another new recombinant plasmid pIRES-Sj26-Sj 14. The expression of Sj14 and Sj26genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj 14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and pIRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj14-Sj26 that contains fatty binding protein (Sj14), GST (Sj26) and paramyocin (Sj97) that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was trans-fected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and pIRES-Sj97-Sj14-Sj26 plasmid DNA, pIRES-Sj14-Sj26 plasmid DNA, plRES-Sj26 plasmid DNA,plRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh.Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the pIRES-Sj97-Sj14-Sj26 group as compared with the plRES blank vector, normal saline and pIRES-Sj26 groups (P<0.01) and the pIRES-Sj14-Sj26(P<0.05). Single splenocyte suspension was prepared to detected the level of IFN-γ by ELISA and the lymphocyte stimulating index (SI) by MTT SI was significantly higher of in the pIRES-Sj97-Sj14-Sj26 group than in other groups (P<0.01), while the IFN-γ, level was significantly higher the pIRES-Sj97-Sj14-Sj26 group than in pIRES blank vector and normal saline groups (P<0.01), but no significant differences were found when compared with pIRES-Sj14-Sj26 and pIRES-Sj26 groups. Flow cytometery showed that the percent-ages of CD4+ and CD8+ T cells were much higher in the pIRES-Sj97-Sj14-Sj26 group (P<0.01,P<0.05). It was concluded that pIRES-Sj97-Sj14-Sj26 vaccine may induce stronger immune response in BALB/c mice.