Objective To investigate the effect of p53-gene expression on renal function in rat with low birth weight ( LBW) .Methods Fifteen pregnant rats were randomly assigned into normal group, LBW group, and L-arginine ( L-Arg)-treated group.For normal group, pregnant rats were fed with 21% protein diet during pregnancy.For LBW and L-Arg treated groups, rats were fed with 10%protein diet.After de-livery, all rats were fed with 21%protein diet.For L-Arg-treated group, rats were given a supplementation with L-Arg (200 mg/kg) drinking water during 21 d lactation, other rats were given running water.At the age of 2m, the ultrastructural change in mesangial cell and podocyte was observed with electron microscope. At the age of 7 d, 21 d, 2 m, and 3 m old, the p53 mRNA expression in renal tissues was observed with re-verse transcription polymerase chain reaction ( RT-PCR) , blood and urine were collected to detect biochem-ical indicators of renal function and 24 h-urine protein.Results ⑴At the age of 3 m,the blood urea nitro-gen ( BUN) and urine cretinine ( UCr) in normal group were significantly lower than those in LBW group ( P <0.01).Compared to normal group, the Ucr of LBW group was significantly lower at the age of 2 m ( P<0.05).At the age of 3 m,the Ucr of L-Arg treated group and LBW group was significantly lower ( P <0.05), the level of 24 h-Urine protein was notably increased in LBW group and L-Arg treated group than that in normal group ( P <0.01, P <0.05) .⑵At the age of 3 m,the expression of p53 mRNA in LBW group was higher obviously than that in normal group.There is a significantly negative correlation between the expression of p53 mRNA and the level of Ucr in LBW group ( r =-0.91, P <0.05).⑶ There were mesangial cell proliferation with matrix increase, filtration membrane podocyte reduction, and partially dis-solved in LBW group, the mesangial cell proliferation of L-Arg treated group was decreased compared to that in LBW group.Conclusions The higher expression of p53 gene in LBW group might be one of reasons for the decreased renal function in LBW rats.

Objective To evaluate the impacts of comprehensive diabetes intervention to provide evidence for effective diabetic control in rural communities.Methods Diabetic patients from four communities at Wujin District of Changzhou City were selected.One hundred and seventy-three patients from two communities received comprehensive intervention,including medication therapy,diabetes education,and diet.exercise,and behavior instruction(intervention group);142 patients from the other two communities got medication therapy only (control group).All the participants were interviewed with structured questionnaire and phyrsical examination.The second epidemic investigation was conducted at 4 years to evaluate the effects of the intervention.Results At the end of the intervention,the awareness rates of normal fasting plasma glucose,major diabetes risk factors,healthy diet,and therapeutic strategies in the intervention group were 90.8%.74.6%,89.6%,and 59.5%,respectively,which were significantly higher than those in the control group (all P<0.05).For the intervention group,the rates of diet behavior or physical exercise change were 68.8% and 58.4%.respectively.which were much higher than those of the control group(all P<0.05).The average levels of systolic pressure,diastolic pressure,fasting plasma glucose,and total cholesterol were (130.6±10.4)mm Hg(1 mm Hg=0.133 kPa),(78.4±7.5)mm Hg,(6.2±3.1)mmol/L,and (4.5±0.9)mmol/L,respectively,in the intervention group,which were lower than those in the control group(all P<0.05).Conclusion Comprehensive intervention shows significant effects on rural community patients with diabetes.

Objective To study the effect of renal p53 protein expression on kidney development in rats with intrauterine growth restriction ( IUGR) . Methods Pregnant rats were randomly assigned into normal group, IUGR group and L-Arg treated group. Normal group were fed with normal diet (21％protein). IUGR group and L-Arg treated group were fed with low-protein diet (10％ protein). During lactation, maternal rats in the three groups were all fed with normal diet. Maternal rats in L-Arg treated group were given additional special drinking water ( L-Arg:200 mg/kg) , while maternal rats in normal and IUGR group were given normal drinking water. Offspring weaned after 21 d and had free access to normal rodent diet and water. When the pups grew up to 2 m, the number of glomeruli was counted using acid digestion method, the proliferation and apoptosis of glomerular and tubular cells were studied using TUNEL and Ki-67 immunohistochemistry, and the ultrastructure of epithelial cells was determined by electron microscopy. At 7 d, 21 d, 2 m and 3 m, p53 protein expression in renal tissue was measured by Western blot, respectively. Results At 2 m, the number of glomerulus ( right kidney) in IUGR group was significantly lower than normal group [(23 647±541) vs. (27 689±492), P＜0. 01]; the index of renal cell apoptosis in IUGR group was higher than normal group [(21. 9 ± 2. 0) vs. (16. 7 ± 2. 5), P＜0. 05];however, IUGR group and normal group had no statistically significant difference (P＞0. 05). The proliferation of mesangial cells was found in IUGR and L-Arg treated group, but not in normal group. And the extent of proliferation in L-Arg treated group was lesser than IUGR group. A reduction of foot process and partial fusion of foot process could be observed in IUGR group and L-Arg treated group while the foot process morphology in normal group was normal. At 2 m and 3 m, p53 protein level in IUGR group and L-Arg treated group were higher than normal group [ 2 m: ( 0. 28 ± 0. 03 ) and ( 0. 21 ± 0. 01 ) vs. (0. 10±0. 02);3 m:(0. 39±0. 04) and (0. 26±0. 02) vs. (0. 17±0. 03);P＜0. 01], while IUGR group higher than L-Arg treated group ( P＜0. 05 ) . Conclusions The kidney of IUGR rats showed reduced glomerular number, increased renal cell apoptosis, enhanced p53 protein expression, increased proliferation of glomerular mesangial cells, decreased foot process, and partial fusion of foot process. And L-Arg could to some extent improve the organizational structure of the kidney in IUGR rats.

Objective To determine the best extraction process of ginkgo leaves with the total transfer rate of total flavonol glycosides and terpene lactones as the index.Methods The effects of ethanol concentration, solid-liquid ratio and extraction time on extraction process were investigated by orthogonal design method, and the contents of total flavonol glycosides and terpene lactones were detected by HPLC to calculate transfer rate.Results The optimum extraction conditions were as follows:85% ethanol refluxing and extracting for three times;the first time extracting with five-fold amount of solvent (V/W) for 3 hours;the last two times with three-fold solvent (V/W) for 2 hours.Conclusion This extraction process has the advantages of simplicity of operator, reason, energy conservation, high efficiency, and is suitable for industrial production.

Objective To develop an effective and broad immune protective vaccination strategy by using DNA and recombinant vaccinia-based H5N1 vaccines.Methods BALB/c mice were immunized with various prime-boost regimens by using different DNA ( pIRES-based or pVRC-based) and recombinant vaccinia (Tiantan strain, rTTV)-based H5N1 vaccines expressing multivalent antigens (HA, NA, M1 and M2).The differences of immunity induced by two DNA vaccines were compared between intradermal electro -poration (IDE) and intramuscular electroporation (IME) deliveries.Immune responses were analyzed by hemagglutination inhibition( HAI) assay, neuraminidase ( NA)-specific antibody measured by ELISA , mi-croneutralization assay and IFN-γELISPOT assay .Results High levels of humoral immunity and T cell re -sponses were induced in mice primed with DNA-based vaccine than those primed with rTTVb-ased vaccine . DNA priming by IDE resulted in higher levels of neutralizing antibody in mice than those by IME delivery . Higher levels of HAI and anti-NA antibodies as well as NA-specific T cell responses were induced by pVRC-based DNA prime than those by pIRES-based DNA prime .HA-specific T cell responses were also enhanced in mice primed with pVRC-based DNA than those primed with pIRES-based DNA by IME .Conclusion The prime-boost strategies by using DNA-based vaccine in combination with rTTV-based H5N1 vaccine could induce humoral and T cell responses in mice against multi-antigens .Immunities induced by vaccines in com-bination might be modulated by various prime regimes .The study provided references for the further develop-ment of novel H5N1 vaccine and the optimization of immunization programs of combined multi-antigen vac-cine candidates .

Objective To better characterize the clinical and histopathological manifestations, diagnosis and differential diagnosis of epithelioid sarcoma (ES).Methods Clinical data were collected from nine patients with ES diagnosed and treated at the Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College from 2000 to 2014.The clinical and histopathological manifestations, immunophenotype, treatment and prognosis of ES were reviewed retrospectively.Results The median age at onset of ES was 34.5 years in these patients, and the average disease duration was 8.3 years.Lesions began on the extremities in 8 patients, and manifested as nodules,ulceration and lymphedema.Histopathological examination showed that tumor cells mainly consisted of spindle cells and epithelioid cells, some of which were in a palisade arrangement with central necrosis.Cytokeratin, epithelial membrane antigen (EMA) and vimentin were coexpressed by tumor cells.Recurrence was observed at the original site in 6 patients after lesion resection, and pulmonary metastasis occurred in 4 patients.Five patients were followed up and two of them died of pulmonary metastasis.Conclusions Local recurrence is frequent in patients with ES after lesion removal.Lymph node and pulmonary metastases of ES are common, and pulmonary metastasis is usually associated with a poor prognosis.

Objective To construct a reverse genetic platform for influenza B virus and to rescue influenza B virus.Methods Eight plasmids carrying the gene segments of B/Florida/4/2006 virus were constructed by using the bidirectional promoter vector pHW2000.293T cells were co-cultured with MadinDarby canine kidney (MDCK) cells and then transfected with the eight plasmids.The supernatants of cell culture and cell debris were collected after transfection and then injected into embryonated chicken eggs and MDCK cells for rescuing the influenza B virus strains.Results This reverse genetic system could be used for the preparation of reassortant influenza B virus strains.The titers of hemagglutination units of the rescued virus achieved 128-256/50μl.Most of the reassortant virus particles were spherical under electron microscope.Conclusion The pHW2000 reverse genetic system could be used for the rescue of influenza B virus.Moreover,it could also be used for the construction of influenza B virus with specific mutations for further in vestigation on the characteristics of influenza B virus and the construction of vaccine strain.

Objective To explore the effect of early application of pediauic amino acids on extrauterine growth and development of preterm and small for gestational age infants.Methods A total of 106preterm and small for gestational age infants was selected in our NICU from June 2011 to May 2013,and randomly divided into two groups:treatment group (group T) and control group (group C).Next,each group was divided into two subgroups according to gestational age and birth weight.Such as ≥34W group (group T1,group C1),＜ 34W group (group T2,group C2),＜ 1.5 kg group (group T3,group C3) and ≥ 1.5 kg group (group T4,group C4).They were observed the effect of extrauterine growth and correlative serum biochemical indicators with application of pediatric amino acid in postnatal 6 hours and 12 hours.Resuits After 2-week treatment,the head circumference and weight growth of group T were higher than that of group C (P ＜ 0.05,or P ＜ 0.01),and the time of birth weight recovery was significantly shortened(P ＜ 0.05,or P ＜ 0.01).The incidence rate of extrauterine growth retardation(EUGR) in the group T was lower than that in group C,there is significantly statistical differences(P ＜ 0.01).The concentration of blood urea nitrogen (BUN) in group T was obviously increased than that in the group C(P ＜0.01).The levels of ALT,aspartate aminotransferase (AST),total bilirubin (TBIL),cholesterol (CH) and triglycerides (TG) were not different between the group T and group C.The comparison of alanine aminotransferase (ALT) levels were statistically significant between group C4 and group T4.In the different gestational age groups,the concentration of BUN in group T was obviously higher than that in the group C after 2-week treatment(P ＜0.05),the levels of AST,TBIL,CH and TG were not different between the group T and group C (P ＞ 0.05).The incidence rate of EUGR in the 4th group C was increased significantly than that in the 4th group T(P ＜0.05).Conclusions The preterm and small for gestational age infants were safe after the pediatric amino acid was used at the 6 h after birth.Amino acid can promote growth of head circumference and weight,shorten the birth weight recovery time and reduce the incidence of EUGR.

Objective To develop an effective and broad immune protective H5N1 vaccine.Methods We first developed two recombinant vaccinia ( Tiantan strain) virus ( rTTV ) based H5N1 vaccines, which consisted of bicistron expressing the hemagglutinin(HA) and matrix protein 2(M2), or bicistron expressing the neuraminidase(NA) and matrix protein 1 (M1). The expression of H5N1 protein in rTTVs was confirmed. We immunized the BALB/c mice twice with two kind of dose ( 104 PFU, 107 PFU)using different combination. Subsequently, we assessed the humoral and cellular immune response in vaccinated mice. Results Our data showed that rTTV-based H5N1 vaccine induced rapidly robust HA- and NAspecific antibody level and IFN-γ secreting form cell(SFC) with either single dose of 107 PFU or twice dose of 104 PFU or 107 PFU. We also detected significant neutralizing antibody and matrix-specific immune response. In addition, we found that immunization with two kind of rTTV-based H5N1 vaccines induced much high level of M2-specific antibody than that with single of rTTV-based H5N1 vaccine. Conclusion rTTVbased H5N1 vaccines in this study elicited board array of immunity and our study offers a promising alternative H5N1 vaccine candidates with favorable potential to prevent various H5N1 pandemic.

Objective To investigate the relationship between the clinical manifestations,prognosis and histopathological findings of mucormycosis.Methods The clinical data on and pathological findings from 7 cases of mucormycosis confirmed by fungal culture in the institute from 1989 to 2006 were analyzed retrospectively.Results There was 1 case of hinocerebral mucormycosis and 6 cases of cutaneous mucormycosis,among them,2 were mucormycotic necrotizing fasciitis (MNF).The condition of patients with rhinocerebral mucormycosis or MNF aggravated rapidly and all the 3 patients died from mucormycosis. Histopathological examination showed mixed infiltrates of inflammatory cells as well as necrosis and angioin vasion.On the contrary,the condition of the remaining 4 patients with cutanesus mucormycosis,who presented mainly with indurated erythematous patch,progressed slowly,and 2 patients were cured.Histologically,the lesions were characterized by granulbmatous infiltration with a few hyphae;no typical angioinva sion phenomenon was noted.There was no evidence of perineural invasion with hyphae in any of the 7 cases.ConclusionIn patients with mucormycosis,histopathological findings characterized by mixed infiltrates of inflammatory cells,numerous hyphae and typical angioinvasion phenomenon may herald a poor prognosis.