Objective:To investigate the inhibitory effect of recombinant mutant human tumor necrosis factor(rmhTNF-?)combined with cisplatin(DDP)against transplanted Lewis lung carcinoma(LLC)in mice,and to explore the related mechanism.Methods:LLC cells were subcutaneously inoculated into C57BL/6 mice at the right axilla and the mice were randomly divided into 4 groups:sodium chloride(control group),DDP group,rmhTNF-? group,rmhTNF-? plus DDP group.On the 7th day after inoculation the diameter of the tumor reached 0.6 cm;the corresponding agents were intratumorally injected for 3 d.All mice were sacrificed 1 d later and the tumors were obtained,weighed,and the tumor inhibitory rate was calculated.Flow cytometric analysis was used to examine cells apoptosis and cell cycle.Expression of survivin mRNA was examined by RT-PCR.Results:(1)The tumor inhibitory rates of the DDP plus rmhTNF-? group(36.61%)was significantly higher than those of the DDP group(17.12%)and rmhTNF-? group(15.83%,P

Objective To investigate the current situation of iodine deficiency disorders and the effect of control measures in Sanmenxia city of Henan province, and to formulate targeted control measures for iodine deficiency disorders. Methods In 2008, five villages(offices) were selected according to the east, west, south,north and center position in each county in 6 counties(cities, districts) of Lingbao, Shan, Hubin, Yima, Mianchi,and Lushi in Sanmenxia city. One primary school was selected in each village(office), fifty students aged 8 - 10 in each primary school were randomly selected to check thyroid and their urine and edible salt were collected to detect iodine. Thirty 5th-grade students in each school were selected to carry out the questionnaire survey of health education about iodine deficiency disorders. According to the east, west, south, north and center position, five potable water samples were selected to detect iodine in the village where the primary school was in. Questionnaire survey was also carried out among ten housewives in the village. Thyroid was examined by palpation;ammonium persulfate digestion-arsenic cerium catalytic spectrophotometry(WS/T 107-2006) was used to detect urinary iodine;direct titration(GB/T 13025-1999 ) was used to detect salt iodine;arsenic cerium catalytic spectrophotometry (GB/T 5750.5-2006) was used to detect water iodine. Results One thousand and five hundreds children aged 8 - 10 were analyzed in 6 counties(cities, districts), the rate of goiter was 3.30%(50/1500), the median of urinary iodine was 273.15 μg/L. One thousand and five hundreds salt samples were detected, the coverage rate of iodized salt was 99.93% (1499/1500), the qualified rate of iodized salt was 98.00% (1469/1499), the edible rate of qualified iodized salt was 97.93% (1469/1500), the coverage rate of non-iodized salt was 0.07% (1/1500), the median of salt iodine was 28.9 mg/kg. One hundred and fifty water samples were collected, the median of water iodine was 2.76 μg/L. The pass rate of the questionnaire survey of health education about iodine deficiency disorders was 97.11% (874/900) in students and 98.67% (296/300) in housewives, respectively. Conclusions The prevention and control of iodine deficiency disorders has achieved significant results and the monitoring indicators have reached the national standard for eliminating iodine deficiency disorders in Sanmenxia city.

Objective To establish a mini-column centrifugation-HPLC method to determine the entrapment efficiency of levodopa-loaded PEGylated-solid lipid nanoparticles.Methods A dextran gel(Sephadex G-50) mini-column centrifugation was employed to separate the free drug from solid lipid nanoparticles.The content of levodopa was qualified by HPLC.Results Under the applied chromatographic condition,the excipients had no influence on the determination of levodopa.A calibrated linear of levodopa concentration was within 10.54-527.00 μg·mL-1.The recoveries of high,medium and low concentrations of levodopa were 99.13%,99.51% and 99.04%(RSD were 1.25%,1.91% and 1.71%), respectively.The free levodopa was well separated from solid lipid nanoparticles by using mini-column centrifugation.The addition of blank solid lipid nanoparticles recovery was 98.84% with RSD of 0.80%(n=3).The average adsorption rates of the three concentrations of free levodopa were 100.00%,98.75% and 98.56%(RSD were 0.00%,0.19% and 0.18%,n=3),respectively.The adsorption rate of the physical mixtures of three different concentrations of drugs and empty PEGylated solid lipid nanoparticles were 99.68%,98.46% and 99.21%(RSD were 1.52%,0.23% and 0.21%),respectively.Conclusion The method was simple,accurate and reproducible,which can be used for determination of the entrapment efficiency of levodopa-loaded PEGylated-solid lipid nanoparticles.

AIM:To explore the diagnosis and treatment methods of radioaction-induced optic neuritis ( RION) through the clinical dates of 17 patients.
METHODS: It was a retrospective case series study. From August 2008 to October 2013, 17 cases (24 eyes) of Rion clinical dates from Chinese PLA General Hospital were studied. The diagnosis methods including visual acuity, pupil, fundus, visual field, fundus fluorescein angiography (FFA), visual electrophysiological testing, and head MRI. To analysis the clinical date of patients with diagnosis of RION by statistical description.
RESULTS: The deterioration degree of vision: 13 eyes were classified as Ⅳ, 9 eyes as Ⅲ, 2 eyes as II. Ten eyes RAPD ( + ) , visual electrophysiology is extinguished. The retina of 5 eyes showed flame hemorrhages and cotton wool spots exudation. Optic nerve head edema in one eye. T1 - weighted MRI enhanced in 19 eyes which showed optic nerve of the intracranial and intratubal segments abnormal changed, optic chiasm and pituitary stalk signal abnormalities and enhancement of the optic nerve. Tortuous optic nerves and rough edges were observed in 5 eyes. Treatment effect: 4 eyes of visual acuity improved, 1 eye from blindness to light perception,1 eye from 0. 08 to 0. 2, 1 eye from 0. 4 to 0. 6,1 eye from 0. 04 to 0. 15, the rest of the cases did not see any improvement.
CONCLUSION: The unique clinical manifestation of RION can provide objective basis for clinical diagnosis in time, but there have not been proven any effective treatments.

Phage display technology has been developed as a powerful tool for selecting polypeptides or proteins with desired biological and physicochemical properties from huge random peptide libraries.Fragments of foreign peptides or proteins that are expressed as fused proteins displayed on the phage surface can keep their relatively independent spatial structure and biological activity,so that they can interact with their ligands to mimic selection of specific molecular epitopes,thus providing an efficient high-throughput screening system.Phage display has been used to allow rapid identification of peptide ligands for a variety of target molecules by an in vitro selection process called "panning".Phage display techniques can be widely exploited to construct tumor-associated antibody libraries,select polypeptides tumor-associated antigen,investigate antigen epitope and design vaccines and medicine,and are used especially as a tool for the diagnosis and treatment of tumor,gene therapy and tumor cell signal transduction research.Recent applications and advanced developments of phage display in cancer research are discussed in this article for the further reference to investigators.

Objective To evaluate the biological toxicity of heavy metals by using Caenorhabditis elegans. Methods The C. elegans at L4 stage were exposed to CdCl2 CrCl3 As2O3 PbCl2 HgCl2 with low concentrations and M9 buffer the control group for 72 h respectively and the effects of heavy metals with different concentrations on the survival time and reproduction of C. elegans were evaluated. Results After exposure to 2.5 10μmol/L HgCl2 and PbCl2 10μmol/L CdCl2 and 50μmol/L CrCl3 for 72 h respectively the life spans and survival curves of the C. elegans were different from those in the control group the differences were statistically significant all P<0.05 . After exposure to CdCl2 CrCl3 As2O3 PbCl2 and HgCl2 with the con?centrations of 2.5 50 100μmol/L for 72 h respectively the generational time and brood size of C. elegans were all different from those in the control group all P<0.01 . Among the 5 heavy metals at low concentrations the reproduction toxicity of Hg was bigger than Pb Cd Cr and the toxicity of As was the weakest. Conclusion Heavy metal exposure can affect the life span and reproductive toxicity of C. elegans.

Objective To explore the protective effect of bone marrow mesenchymal stem cells (MSC) on diabetic myocardium and anoxic pre-conditioning (AP).Methods Eight-week-- old male Sprague-Dawley rats were given with a single intraperitoneal injection of streptozotocin (STZ) (60 mg/kg)to induce diabetes mellitus (DM).Donor rats were 8-week-old male Sprague - Dawley rats.Before transplantation,MSC were incubated in CM-DiI at a concentration of 2 μg/mL for 20 min.AP-MSC were exposed to 3 hours of anoxia.At 4 months after STZ injection,diabetic rats were randomly given with an intramyocardial injection of one of the followings:150 μL of Dulbecco's modified Eagle's medium ( DMEM),5 ×106 MSC/150 μL,or 5 × 106 AP - MSC/150 μL (n =10 for each group).Three months after STZ injection and 2 weeks after transplantation,we evaluated the cardiac function by echocardiography,and also evaluated the cardiac conditions by alkaline phosphatase staining,western blot analysis for apoptosis related proteins and signal pathways.Results MSC,especially AP- MSC increased fractional shortening (FS) of diabetic heart (P ＜0.01 vs DMEM respectively).AP-MSC greatly increased the capillary density of diabetic myocardium (P ＜0.01 vs DMEM and MSC group respectively).AP-MSC are anti-apoptotic in the rat DCM model,possibly mediated through cardiac upregulation of Bcl-2/Bax ratio ( P ＜ 0.05 ) and inhibiting the expression and activation of caspase - 3 ( P ＜ 0.01 ).Conclusions Intramyocardial transplantation of APMSC has a protective effect on diabetic cardiomyopathy.

Objective To prepare monoclonal antibody in mice so as to develop an ELISA method for diagnosis of Toxoplasma gondii infection during the initial stage. Methods The mice were immunized by combining routine and intrasplenic immunization with recombinant SGA1 antigen. B lymphocyte hybridization technique was applied to prepare the anti-SAG1 McAbs. Positive clones were screened using ELISA and subcloned to establish cell lines. Ascites was induced to produce the McAbs. Then the McAbs were purified by protein G chromatograph column. The specificity of McAbs was identified by Western blot and sandwich-ELISA. Sensitivity of the McAbs was determined using sandwich-ELISA. Comparasion was carried out between PCR and sandwich-ELISA method. Results Two positive clones were obtained and named as 3B6, 10C4, both could identify the native and recombinant SAG1 antigens. The sensitivity of 3B6, 10C4 was 31.3 ng and 62.5 ng, respectively. There was no cross reaction between the McAbs and positive sera from patients with schistosomiasis, ancylostomiasis or malaria. By using PCR and ELISA, the positive infection rate of T. gondii was 63.2% and 47.4%, respectively. Conclusions Therefore, mouse anti-rSAG1 antigen McAbs have been prepared successfully and primarily applied to early stage diagnosis of T. gondii infection.

This study aimed to investigate the drug susceptibility of wild-type and mutant avian influenza A (H7N9) virus neuraminidase (NA) to oseltamivir and zanamivir. Codon optimized DNA of H7N9 (A/ Hangzhou/1/2013) NA was synthesized and constructed into the pcDNA3.1/His vector (NA(H7N9-WT)). Mutant NA(H7N9-H274Y) and NA(H7N9-R292K) plasmids were constructed by directed mutagenesis PCR using NA(H7N9-WT) plasmid as the template followed by sequencing. NA plasmids were transfected into 293T cells and cell lysates containing NAs were collected 48 h post-transfection. Wild-type and mutant NAs were analyzed by Western blotting and their activities were tested by the 4-MUNANA-based assay. All three NAs were expressed and enzymatic activities were confirmed. The effects of oseltamivir and zanamivir on all three NAs were then tested. It showed that the half maximal inhibitory concentrations (IC50s) of oseltamivir carboxylate on NA(H7N9-WT), NA(H7N9-H274Y) and NA(H7N9-R292K) were 1.6 nM, 15.1 nM, and > 1 000 nM with fold changes of 9 and > 625, respectively. The IC50 values of zanamivir on NA(H7N9-WT), NA(H7N9-H274Y), and NA(H7N9-R292K) were 1.1 nM, 1.4 nM, and 38.0 nM with fold changes of 1.3 and 34, respectively. These results indicated that oseltamivir and zanamivir could significantly inhibit NA(H7N9-WT). NA(H7N9-R292K) showed high-level resistance to both drugs (34-fold and 625-fold) and NA(H7N9-H274Y) was sensitive to both (1.3-fold and 9-fold). These results indicated that both oseltamivir and zanamivir could be used for patients infected with the H7N9 virus. However, when patients carried the H7N9 virus with a NA R292K mutation, other medications would be preferred over oseltamivir or zanamivir.
Antiviral Agents ; pharmacology ; Humans ; Influenza A Virus, H7N9 Subtype ; drug effects ; enzymology ; genetics ; Influenza, Human ; virology ; Microbial Sensitivity Tests ; Mutation ; Neuraminidase ; antagonists & inhibitors ; genetics ; metabolism ; Oseltamivir ; pharmacology ; Viral Proteins ; antagonists & inhibitors ; genetics ; metabolism ; Zanamivir ; pharmacology

Objective To study the relationship of surgical procedures with clinical effect in herniat-ed lumbar disc in order to improve operative methods and obtain a better outcome.Methods Retrospec-tive analysis was carried out in7235patients with herniated lumbar disc,who had been operated by removal of nucleus pulposus using small incision and fenestration in our hospital since1983,313cases of whom re-ceived second operation because of postoperative complications.There were187males and126females aging from27to62years(mean,45.8years),the incidence of morbidity was4.32%.During the same period,552patients,who had less approved operative result primarily treated in other hospitals,were admitted to our hospital.There were317males and235females aging from31to64years(mean,46.0years).Results The postoperative complications could be divided into short term(within1month after operation)and mid-dle to long term(more than1month after operation)groups.The total446cases with short term compli-cation were133cases primarily treated in our hospital and313cases in other hospital,including of re-herni a-tion or incomplete nuclear extirpation in77treated with second operation,discitis in106treated with con-servative therapy for 67and second operative management for 39,canal hematoma compression in76treated surgi cally,multi-level herniation and leakage in21and mislocalization in59re-operated,injury of nerve root or cauda equina in85treated with nerve exploration,release and anastomosis,residual materials in canal in17with removal of foreign body,venous thrombus in2,and arachnoiditis in3.The total424cases with middle to long term complications were185cases primarily treated in our hospital and239cases in other hospitals,including of nerve root adhesion in159,recurred disc herniation in122,segmental instability in81,and ia-trogenic spinal stenosis in62.Conclusion Many factors may influence the outcome of herniated lum bar disc which can be abolished by sufficient preparation,careful operation and proper managememt.