Objective To establish a method for the cultivation of hain follicle outer root sheath (ORS) cells of murine vibrissa and to study their expression of uPA/uPAR. Methods The ORS cells of murine vibrissa were cultured by combination of enzyme digestion and tissue culture with the feeder layer of dermal sheath cells of the hair follicle. The cells and their uPA/uPAR expression were identified by immunocytochemistry (ICC). Results The feeder layer of DS was suitable for the culture of ORS cells. ORS cells expressed uPA/uPAR protein. Conclusion The DS of hair follicle is a better feeder layer for the growth of ORS cells. ORS cells have the ability to migrate and proliferate.

Objective To establish immortalized neural progenitor cell strain and provide stable cell resource for cell-transplantation and gene therapies. Methods Plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigen gene (SV40Tag) were transfected into the primary cultured neural progenitor cells (NPCs) of newborn rat using lipofectin transfection method. Colonies were isolated by puromycin selection and expanded by many passages. Anti-nestin antibodies were used to identify the cultured cells. The specific molecular marker of neurons and astrocytes were detected using immunocytochemistry method to investigate the capability of differentiation of the transfected cells. The expression of SV40Tag in expanded cell lines was identified by RT-PCR, Southern blot and immunocytochemistry method.Results One anti-puromycin cell clone was obtained, which was microtubule-associated protein-2 (MAP-2) positive cells with the capability of proliferation and could differentiate into MAP-2 or glial fibrillary acidic protein positive cells. The existence of SV40Tag cDNA and the expression of mRNA and protein of SV40Tag were confirmed in transfected cells. The transfected cells were expanded to immortalized cell strain maintained for more than 50 passages, named as immortalized neural progenitor cell (INPC) . INPCs were elliptical or triangular cells with two or three short axons. The population doubling time of INPC was (22.9?2.7)h, subculture, freezing and recovering had no effect on cellular shape and proliferation of INPC. Conclusion Immortalized neural progenitor cell strain was established successfully. It may provide stable cell resource for the basic researches and cell-transplantation therapies with NPC.

Objective To explore the roles of urokinase type PA (uPA) and uPA receptor (uPA R) in the course of repair after human embryo corneal alkali burn in vitro . Methods After the alkali burn model in vitro was established successfully, some techniques such as the observation of cell culture and morphology, ICC and image analysis were used. Results No significant difference was found between the cells from corneal limbus and central cornea and almost all of them expressed AE1/AE3. The expressions of uPA and uPA R increased to the maximum at about 24 h after alkali burn. uPA expression distributed mainly in the cytoplasm but uPA R expression distributed mainly in the cell membrane. Conclusion The corneal epithelial cells of comparatively high purity can be acquired by tissue culture. There probably exists difference in development and vitality between embryo and adult cornea. There might be commonness of the roles of uPA and uPA R in epithelial tissue.

Environmental physiological method adopted, the ergonomic experiment is to seek the best parameter of the protective hood structural size, on which the structure design of the protective hood will be based, through the analysis of the organic relationship between the microenvironment of power air-purifying medical protective hood and the body's physiological characteristics. The changes of such indexes due to different air currents are analyzed as the oxygen, carbon dioxide, temperature and humidity in the microenvironment. The changes of testees' physiological parameters and their responses to psychological loads and body strengths are paid attention to. The results show the hood can meet the testees' demands with proper air current.

ATP-binding cassette protein E (ABCE1) has been annotated as an Rnase L inhibitor in eukaryotes. Previous study showed that the overexpression of ABCE1 was related with the occurrence and clinical stage of lung adenocarcinoma. As an initial investigation into the novel functions of ABCE1, siRNA-expressing vectors targeting sites of the ABCE1 gene were constructed from RNAi-Ready pSIREN-DNR-DsRed-Express vector. Cultured 95-D and NCI-H446 lung carcinoma cells were transfected with the siRNA-expressing vectors using FuGENE 6 and transfection efficiency was determined by using fluorescence microscopy. The expression level of ABCE1 protein was determined by Western blot and immunofluorescence staining. Cell viability was determined by MTT, cell cycle was analysed by flow cytometry.The apoptotic rate was observed by ELISA. Fluorescence microscopy showed a satisfactory transfection efficiency which was about 42.70%. Cell viability and the growth fraction were markedly suppressed, whereas the apoptosis was significantly increased in SiRNA-95-D and SiRNA-NCI-H446 cells than controls(P< 0.05). It can be concluded that the siRNA targeting ABCE1 gene shows a dramatic inhibitory effect on RNA transcription and protein expression and a promoting effect on the apoptosis in 95-D/NCI-H446 cells, which offers a reliable base for the further in vivo experiment.

Objective To investigate the surgical procedure selection for chronic pancreatitis.Methods The clinical data of 80 patients with chronic pancreatitis who were admitted to the Peking University First Hospital from January 2000 to August 2013 were retrospectively analyzed.Thirty-eight patients were with or without pancreatic duct stone,and the dilation of the pancreatic duct was above 7 mm,44 patients were with common bile duct dilation,32 patients were with inflammatory mass in the head of the pancreas,and 3 patients were with splenomegaly and esophagogastric varices.Surgical procedures were selected according to the symptoms and results of imaging examination.The remission or recurrence of pain was judged according to the visual analog scales.Patients were followed up via out-patient examination,mail or phone call till December 2013.Results Choledochojejunostomy was done on 27 patients,Partington-Rochelle pancreaticojejunostomy on 24 patients,PartingtonRochelle pancreaticojejunostomy + choledochojejunostomy on 6 patients,pancreaticoduodenectomy on 7 patients,resection of the body and tail of the pancreas on 4 patients,Beger's procedure on 3 patients,splenectomy on 3 patients,Frey's procedure + fenestration of bile duct in the head of the pancreas on 3 patients,Frey's procedure on 2 patients,common bile duct exploration + T tube drainage on 1 patient.The remission rate of abdominal pain was 95.2％ (60/63).One patient died of abdominal infection and multiple organ dysfunction syndrome perioperatively.Three patients were complicated with abdominal infection,2 with pancreatic fistula,1 with biliary fistula and 1 with abdominal bleeding.All the complications were cured by conservative treatment.Seventy-nine patients were followed up,and the mean time of follow-up was 58.6 months (range,4-156 months).Thirty patients had recurrence or new onset of abdominal pain,and the recurrence rate was 38.0％ (30/79).Of the 32 patients with inflammatory mass in the head of the pancreas,17 received choledochojejunostomy,and the recurrence rate of abdominal pain was 9/17 ; the other 15 patients received pancreatoduodenectomy,Beger' procedure or Frey's procedure,and the recurrence rate of abdominal pain was 1/15.Of the 41 patients without inflammatory mass,10 received choledochojejunostomy,and the recurrence rate of abdominal pain was 7/10; 30 received PartingtonRochelle pancreaticojejunostomy,and the recurrence rate of abdominal pain was 33.3％ (10/30).Conclusions Complete drainage could relieve the symptoms for patients with pancreatic duct dilation.Surgical resection or combined surgical procedure is effective for the treatment of patients with inflammatory mass in the head of the pancreas.

Objective To investigate the risks and benefits of different surgical treatments for cystic neoplasms of the pancreas (CNP).Methods The clinical data of 243 CNP patients were reviewed retrospectively.Different surgical treatments were adopted according to the site,size and invasiveness of the tumors.A long term follow-up was carried out for patients with small benign CNP,and a surgical excision is proposed if tumors progressed during the observation.Results 58 outpatients with no evidence of malignancy was followed up and had long-term survival,in which 4 patients received a surgical resection in case of tumor progression,and all of them were confirmed benign tumors.185 cases received surgical treatments,with a resection rate of 97.3％ (180/185),including 127 non-invasive tumors,and 58 cases of invasive tumors.Perioperative mortality was 2/185,and morbidity rate was 41/185.Pancreatic fistula was the most frequent complication,which was significantly associated with tumor site and excision extension.All patients with non-invasive CNP acquired a long term survival after surgical treatments.The postoperative 1,3,5 year survival rates for patients with invasive lesions were 89.6％,52.1％ and 29.2％,respectively.Conclusions Long term follow-up and observation is feasible for asymptomatic patients with benign CNP.A radical resection should be performed for malignancy,and a combined multi-organ resection may improve the prognosis for local advanced tumors.

Objective To determine normal reference ranges for venous blood count among children aging from 1 year old to 12 years old.Methods These normal reference ranges were defined in a population of 526 healthy children who had no blood system diseases,allergic diseases,respiratory system diseases,urinary system diseases,digestive system disease,rheumatoid disease,thyroid disease,parasitic infections,malignancies and genetic disease,etc.Values of white blood cell count (WBC),red blood cell count (RBC),hemoglobin (Hb)concentration,red blood cell specific volume (Hct),mean corpusular volume(MCV),mean cell hemoglo-bin (MCH),mean corpuscular hemoglobin(MCHC),platelet (PLT),percentage of neutrophil (NE%),percentage of lymphocyte (LY%),percentage of mononuclear cells (MO%),percentage of acidophilic granulocyte (EOS%).Statistical analysis was done on various parameters that we recorded,and then for every parameter,we could get the various reference ranges for different age groups.Results The subjects were divided into 4 groups based on age.Besides the parameters of WBC count and classification of WBC,the rest of parameters were proved to be of no statistical difference between 4 groups..After an integration of the values,we could get the results as follows:RBC(4.02-5.2)×10 1 2/L,HGB 108-144 g/L,Hct 35.2%-40.4%,MCV 74.6-89.9 fL,MCH 20.9-34.7 pg,MCHC 332- 340 g/L,PLT(157 - 409 )× 10 9/L.WBC count did not have statistical difference between the age group 6-<9 and 9-12,but did have between the rest groups.After an integration of the values of WBC count,it could be conclu-ded that WBC count of age group1-<3 was(4.88-13.38)×10 9/L,that of age group 3-<6 was(4.26-1 1.6)×10 9/L and that of age group 6-12 was (4.24-10.24)×10 9/L.WBC classification results were various in different age groups.The values showed as follows:age group 1-<3 NE:29%-32%,LY:58%-61%;age group 3 -<6 NE:43%-46%,LY:43%-46%;age group 6-<9NE:49%-52%,LY:38%-40%;age group 9 to 12NE:5 1% - 58%,LY:33% - 39%.Conclusion WBC classification re-sults and WBC count do have statistical difference in different age groups.Besides the parameters of WBC count and classification of WBC,the rest of parameters are proved to be of no statistically difference in different age groups.The values of WBC count decrea-ses as the age increases.From WBC classification results,the most apparent fact is that the percentage of neutrophil increases as the age increases but the percentage of lymphocyte is just the contrary.As mentioned above,we suggest that we should establish a spe-cific whole blood count normal reference range for each age group during our laboratory testing work.

Two hundred and fifty patients with subclinical hypothyroidism ( SCH) and 120 euthyroid volunteers were recruited for the study, SCH patients were stratified into 2 groups according to TSH levels(group A:TSH<10 mIU/ L; group B: TSH>10 mIU/ L). All subjects were examined for clinical characteristics, thyroid profile, lipid profile, and biomarkers of early atherosclerosis. Patients in group B received L-thyroxine replacement treatment to achieve euthyroidism. After 6 months of stable euthyroidism all measurements were repeated. SCH patients had higher levels of total cholesterol(TC), low-density lipoprotein-cholesterol(LDL-C), asymmetric dimethyl arginine (ADMA), carotid artery intima media thickness(CIMT), thromboxane B2(TXB2), and C-reactive protein(CRP) but with lower nitric oxide(NO) level compared with euthyroid subjects. Levels of TC, LDL-C, CIMT, TXB2, and ADMA decreased, and NO level increased significantly after 6 months of euthyroidism. TSH was positively correlated with TC, LDL-C, CIMT, ADMA, and TXB2; and negatively correlated with NO. Based on multivariate regression analysis, TSH was an independent influential factor of CIMT and NO. We conclude that raised TSH is an important risk factor of atherosclerosis in SCH.

Cancer pain is the most important factor affecting the cancer patients' quality of life, and the approach to relieve and control cancer pain is becoming the focus. Pain mechanism research can offer solutions to pain treatment, such as blocking the happening and conduction of analgesia. The earliest μ, κ, σopioid receptors were found in the research of morphine and opioid peptides, especially μ receptor's leading role in pain treatment. Currently, μ opioid agonist is basically used in clinical pain treatment. Morphine, the third level drug, is still the classic pain therapy drugs. Novel drugs such as fentanyl transdermal and controlled-release oxycodone provide new ideas for the pain ease. Opioid combined with non-opioid drugs, the change of opioid drugs delivery way and joint application of controlled release drug and relievers, have dramatically reduced opioid drugs' side effects.