This article reviews the resultsof the basic research about epidermal growth factor and its receptor,and the development of the novel drug, EGF eyedrop, that containing chemically synthesised EGF gene,the construction of EGF expression vector, the transformation of the host cells , the purification of the recombinant protein EGF, the preparation of three batches of the EGF product and identification, the preclinical and clinical trials. Relevant studies show that recombinant EGF consisting of 51 amino acids can be secreted into the medium under the control of the α factor leading sequence in the yeast cells.The EGF can accelerate the growth of corneal-limbal epithelial cells and the healing of an alkali burned corneal. The EGF can be used in curing oral cavity ulcer and skin burned wound. And it has the preventive effects on experimental duodenal ulcer of rat. The antiserum was made for test of the con- centration of blood EGF and urine EGF by RIA. Data from studies demonstrate the inhibition effect of EGF on the growth of tumor cells, such as A431 and BT325 cells in the presence of high EGF concentration (＞10 ng/ml). The expression of EGFR and DNA ploidy in renal carcinoma has clinical significance. Crystallization and preliminary x-ray diffraction studies of the EGF has been made. The MW of the EGF product is 6000, and the pI is about 4. 6 and it has correct N-terminal amino acids sequences , immunogenicity and biological activity. There is no vestige of the DNA of the yeast cells. Animal experiments reveal that there is no cumulation of the EGF in the body, and EGF can promote corneal epithelial healing. There is no toxicological effect during cornea wound healing of rabbit. A randomized, double-blind, placebo-controlled, multi-center clinical trial was conducted in four hospitals to assess safety,ocular tolerance and efficacy of an ophthalmic solution of EGF for 200 cases of cornea transplantation and 247 cases of nebulae. Unequivocal results were obtained as the eyedrop really accelerate the wounded cornea healing. So, the EGF eyedrop as a novel drug of class I is approved by the National Drug Administration, and this is the first gene engineering drug that come from yeast expression system in China.

Aim To investigate the effect of DHA on NGF-induced neuronal differentiation of PC12 cells and explore the possible mechanism via regulating BMP pathway. Methods PC12 cells were treated with 100μg·L-1 NGF and 100 μg·L-1 NGF + 10 μmol· L-1 DHA for 3, 6 and 9 days respectively. The length and number of neurite were detected by immunofluores-cenc. DHA content was analyzed by gas chromatogra-phy in all groups. The protein expression of BMP4, BMP7 , BMPR-II and p-Smad 1/5/8 was determined by Western blot. Results The length of total primary neurite in NGF+DHA groups was obviously increased, longer than that in NGF group; DHA content in 10μmol · L-1 DHA group was higher than that in the control group;NGF+DHA groups also unregulated the protein expression of BMP4 , BMP7 , BMPR-II and p-Smad 1/5/8 . Conclusion DHA promotes NGF-in-duced neuronal differentiation in PC12 cells, which may be associated with the upregulation of BMP path-way protein.

To explore the optimized dispatch of the rescue resources based on the mastered resources informa-tion. The objective function was set as the shortest rescue time, and 0-1 programming models were established and solved for the conditions that the number of medical rescue units was more or less than needed by the missions. In the two conditions, the model's calculated results were both optimal, and could be used to obtain the optimal dispatch solutions while the medical rescue time was the shortest. The 0-1 programming model can be effectively used to optimize dispatch strategy for medical rescue team.

In this study, five rhesus macaques were inoculated intravenously with SIVmac251 to establish a model of simian autoimmune deficiency syndrome (SAIDS). Peripheral blood samples were collected at different time points to monitor changes in the total T cell number and T lymphocyte subset. Plasma viral loads, cytokine expression levels and anti-SIV antibody levels were also assayed to acquire certain basic indexes to evaluate disease progression in the rhesus macaque SAIDS model. During the acute stage of infection, plasma viral loads reached a peak at week 1 post-inoculation and lasted for approximately 3 to 44 weeks. The CD3+ CD4+ T lymphocyte count in peripheral blood also transitorily decreased. During the same period, the level of interferon-gamma show an increasing trend, whereas IL-12 levels decreased; IL-2, IL-4, IL-10 and TNF-alpha were maintained at normal levels or could not be detected. During the asymptomatic and ARC phases, plasma viral loads persisted above 10(4) RNA copies/mL and either increased or declined during the later stages of disease; CD3+ CD4+ counts showed a steadily declining trend and the ratio of CD4 to CD8 decreased during late-stage disease. Moreover, antibodies against viral proteins were detected in the plasma and showed a significant increasing trend, while there were no apparently changes in the levels of IFN-gamma, IL-12, IL-2, IL-4, IL-10 and TNF-alpha. In conclusion, the characteristics of the SIV animal models in our study are similar to those of patients with AIDS. Therefore, the rhesus macaque SIVmac251 infection models can be applied for further studies into AIDS.
Animals ; Antibodies, Viral ; blood ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; virology ; Cytokines ; genetics ; immunology ; Disease Models, Animal ; HIV Infections ; genetics ; immunology ; virology ; HIV-1 ; physiology ; Humans ; Macaca mulatta ; Male ; Simian Acquired Immunodeficiency Syndrome ; genetics ; immunology ; virology ; Simian Immunodeficiency Virus ; physiology ; Viral Load

Murine norovirus (MNV) was first discovered in mice in 2003. MNV is a member of the genus Norovirus in the family Caliciviridae. It is one of the most important and prevalent pathogens of laboratory mice, and almost all mouse strains are susceptible to MNV infection. In this study, a MNV strain was isolated from the cecal contents of infected mice and identified by the cytopathic effect (CPE) assay, virus plaque assay, 50% tissue culture infectious dose (TCID50) assay, electron microscopy, indirect immunofluorescence assay (IFA) and nucleotide sequencing. On infection, the RAW264.7 cell line showed obvious cytopathic effects within 24 to 48 hours post-inoculation, as infected cells became rounded, bright and shrunken, with ultimate disintegration of the cell sheet. After the isolation of the MNV virus, the virus was plaque-purified in RAW264.7 cells. The TCID50 of the virus was 10(5.25/0.1 mL. Electron microscopic observations of the purified virus showed the presence of spherical and non-enveloped viral particles that were 30 to 35 nm in diameter. According to the identification results, the isolate was named as MNV Guangzhou/K162/09/CHN. Thereafter, five overlapping gene fragments that covered the entire open reading frame (ORF) were amplified by RT-PCR, and the 3'-untranslated region (UTR) and 5'-UTR were amplified using the 3'-rapid amplification of cDNA ends (RACE) and the 5'-RACE method, respectively. Each of the gene fragments were cloned and sequenced, and whole genome sequences of the strain were obtained by assembling the cDNA fragment sequences. The results showed that the length of the complete genome was 7 380 nucleotides (GenBank accession number: HQ317203). The comparison of nucleotide and deduced amino acid sequences of the isolate was performed against other MNV strains in the GenBank database. A phylogenetic tree based on VP1 nucleotide sequences was constructed using MEGA5.0 software. The homology of nucleotides between the MNV Guangzhou/K162/09/CHN strain and other MNV isolates ranged from 87.4% to 89.7%. Phylogenetic analysis showed that there was a close genetic relationship between the Guangzhou/K162/09/CHN strain and MNV strains isolated from Japan (S7-P2 and S7-PP3 isolates), Korea (K4 isolate), and Germany (Berlin/04/06/DE and Berlin/05/06/DE isolates). This is the first report of the isolation and identification of MNV in China, and the first report of the genetic analysis of its complete genome.
Animals ; Caliciviridae Infections ; veterinary ; virology ; Mice ; Molecular Sequence Data ; Norovirus ; classification ; genetics ; isolation & purification ; physiology ; Open Reading Frames ; Phylogeny ; Rodent Diseases ; virology ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics

Objective To investigate the role of pre-chemotherapy hemoglobin and platelet levels in the effect of chemotherapy and prognostic outcome in patients with International Federation of Gynecology and Obstetrics(FIGO) stage Ⅰ b2 - Ⅱb cervical cancer treated with neoadjuvant chemotherapy followed by radical hysterectomy.Methods From January 1999 to December 2010,111 patients with FIGO stage Ⅰ b2 - Ⅱ b who underwent chemosurgical treatment at the department of obstetrics and gynecology in Peking Union Medical College Hospital were reviewed.The median age of patients was 42 years (range:21 -68 years).The median level of prechemotherapy hemoglobin and platelet levels was 127 g/L and 266 ×109/L,respectively.Chemotherapy response was evaluated according to the WHO criteria,including complete response (CR),partial response (PR),arable disease (SD) and progressive disease (PD).Patients who achieved CR or PR were defined as responder.Rates of clinical response were compared with the clinicalpathological variables using chi-square test.Multiple logistic regression was carried out to evaluate the relationship among the probability of achieving an optimal clinical response and the variables.The log-rank test was used to compare the homogeneity of progression-free survival and overall survival functions across strata defined by categories of prognostic variables.The Cox proportional hazard model was used to assess the significance of potential prognostic factors for progression-free survival and overall survival.Results All patients received one to three cycles of chemotherapy.After the neoadjuvant chemotherapy,9 patients achieved CR,77 patients PR,23 patients SD, 2 patients PD.The overall response rate was 77.5％(86/111).By univariate analysis,the clinical response rate was associated with tumor grade( P =0.026),deep cervical stromal invasion ( P =0.029 ) and positive lymph nodes ( P =0.048 ).By multiple logistic regression,deep cervical stromal invasion ( P =0.015 ) and positive lymph nodes ( P =0.031 ) were independent predictors of optimal clinical response.By log-rank test,5-year overall survival rate and 5-year progression-free survival rate were associated with lymph nodes metastases status and lymphovascular invasion ( P =0.000),but not with hemoglobin and platelet levels( P ＞ 0.05 ).By Cox regression model,lymph nodes metastases status and lymph-vascular space involvement ( P ＜ 0.01 ) were independently prognostic factors of 5-year overall survival rate and 5-year progression-free survival rate.Conclusion Pretreatment hemoglobin and platelet levels were neither predictors of clinical response to chemotherapy nor prognostic factors.

Objective To investigate the effects of in situ venous arterialization on extensive artery obliterans occlusion of the lower extremity. Methods Lumbar sympathetic ganglionectomy and one stage in situ arterialization of the great saphemous vein were performed in 104 ischemic limbs of 88 patients with extensive arterial occlusion. Results Eighty-two of 104 limbs were followed-up from 6 months to over 6 years. The intermittent claudication, night pain improved in all cases, with satisfactory wound healing and no swelling of the lower limbs. Conclusions Arterial blood flow through venous conduit improves and reconstructs the blood circulation of the ischemic limbs.

To analyze the reasons of impaired vision after LASlK and explore its preventive treatment measures preliminarily.METHODS: ln this retrospective study, 175 eyes of 134 patients whose vision was decreased after LASlK were included. The constituent ratio of every reason was counted and uncorrected visual acuity ( UCVA ) between pre-treatment and post-treatment were compared by paired t-test respectively.RESULTS:The overall incidence of impaired vision after LASlK was 1. 86%. The constituent ratio of regression was 51. 43% and UCVA increased from 0. 61±0. 22 to 0. 90±0. 38 (t=8. 00, P<0. 001) after treatment. The constituent ratio of punctate corneal epithelial defect was 32. 57% and UCVA increased from 0. 60±0. 19 to 1. 20±0. 24 (t=20. 00, P<0. 001 ) after treatment. The constituent ratio of accommodative spasm was 5. 14% and UCVA increased from 0.76±0. 21 to 1. 32±0. 22 (t=8. 14, P<0. 001) after treatment. The constituent ratio of corneal flap shift and gauffer was 4% and UCVA increased from 0. 29 ± 0. 26 to 1. 24 ± 0. 28 ( t = 6. 33, P<0. 001 ) after treatment. The constituent ratio of corticosteroid - induced ocular hypertension was 4% and UCVA increased from 0. 57±0. 05 to 1. 0 ± 0. 16 ( t= 2. 53, P<0. 05 ) after treatment. The constituent ratio of fundus lesions and diffuse lamellar keratitis ( DLK) was 2. 86% and UCVA all increased by different degrees after treatment.CONCLUSlON: The reasons of impaired vision after LASlK are many and varied. These cases could recover their vision by discovery and treatment in time, and the appropriate preventive measures were essential.

Objective To assess the liver histopathological characteristics of chronic hepatitis B (CHB) patients with alanine aminotransferase(ALT)lower than 2 times the upper limit of normal (ULN) through liver biopsy, and try to provide subjective evidence for clinical anti-viral treatment.Methods From October 2005 to August 2010, patients accepted liver biopsy in department of infectious disease, Sichuan provincial people's hospital were enrolled. The criteria for liver biopsy was as follow, (1) HBsAg-positive for more than 6 months, (2) HBeAg-positive patients with HBV DNA ≥103 copies/ml or HBeAg-negative patients with HBV DNA≥ 104copies/ml, (3) ALT was lower than 2 times ULN for more than 6 months,and without any hepatic protectants, (4) never accepted any antiviral treatment before, including IFN or nucleoside analogues, (5) willing to accept liver biopsy. Before liver biopsy, routine blood test, prothrombin time, liver function test, hepatitis B antigen and antibody test, HBV DNA quantification were examined. The biopsy position was located under routine ultrasound, liver biopsy were performed to assess the grading of inflammation and necrosis and the degree of fibrosis. The correlation between all the factors and liver inflammation and fibrosis were analyzed. Results Totally 383 cases (240 males and 143 females) met the diagnostic criteria, aged from 16 to 59 years old and the mean age was 28.0 years old. Cases of liver inflammation in G0, G1, G2, G3andG4 grade was 2 cases (0.5％), 165 cases (43.1％), 191 cases (49.9％), 25 cases (6.5 ％ ) and 0 cases (0 ％ ) respectively, cases≥G2 grade accounted 56.4 ％ of total. Meanwhile,stage of fibrosis in S0, S1, S2, S3 and S4 was 103 cases (26.9％), 265 cases (69. 2％), 13 cases (3.4％), 2 cases (0.5％) and 0 cases (0％) respectively, percentage of liver fibrosis in S2stage and over was only 3.9％. The occurrence of serious liver inflammation was associated with age, ALT levels, HBV DNA levels and HBeAg status (P＜0.05). There was no obvious association between HBV DNA level and liver fibrosis (P＞0.05). Conclusions There were obvious liver inflammation and different degree of liver fibrosis in CHB patients with alanine aminotransferase(ALT)lower than 2 times ULN. The degree of liver injury assessed by liver biopsys is recommended as an evaluation for the necessary of anti-viral therapy.

Objective To establish a Juema minipig model of myocardial infarction, to evaluate the clinical indi?ces in the model pigs, and to explore the relationship between gene expression and metabolic decompensation. Methods 13 male Juema minipigs were randomly divided into control (Sham, n=5), myocardial infarction (MI, n=5) and normal control (for evaluating the recovery condition after surgery, n=3) groups. In the MI group, the ligation was done at the left descending coronary artery around the 1/3 distance to heart apex. Four weeks after the surgery, the cardiac function and serum biochemistry was analyzed. The histological changes and gene expression profiles in the myocardium in the peri?infarct area were exanimated. Results Ultrasonic images showed that the infarction was formed, the ejection fraction and fraction shortening were significantly reduced in the MI group ( ~32% and ~40% less than those of the sham group). Histological examination showed that myocardial fibers at the peri?infarct area were broken, dissolved, and there was con?nective tissue hyperplasia with increased neutrophil and lymphocyte infiltration. Microarray analysis revealed that two myo?cardial remodeling and pathology mediating pathways, three inflammation?related pathways, and 8 metabolic pathways ( in?cluding fatty acid, amino acid, and glucose metabolic pathways) were significantly changed. Conclusions We have suc?cessfully established a Juema minipig model of myocardial infarction. The less branches of the left descending coronary ar?tery allow us to establish a stable model by surgery with comparable characteristics in the clinic indices. The results of this study provides useful reference characteristics of an animal model with characteristic changes in the peri?infarct area.