The bioreactor production of recombinant Lateolabrax japonicus growth hormone (rljGH) expressed intracellularly by Pichia pastoris was investigated. A strategy of feeding methanol at the exponential rate was established and the effect of specific growth rate on the rljGH production was examined. The results indicated that the average specific production rate increased and the rljGH production duration decreased as the specific growth rate increased. The maximum specific rljGH production (0.58 mg/g WCW) was achieved at a specific growth rate of 0.029/h. The effect of supplementing ammonium sulfate, peptone and yeast ex- tract on the rljGH production was further investigated. The results indicated that the effects of ammonium sulfate and peptone were not significant. Supplementing yeast extract of 2.5 g/L was advantageous for the rljGH production. The duration of the rljGH production was increased to 23 h from 17 h and the fermenta- tion stability of run-to-run could be improved.

Objective To investigate the application effect of homemade multifunctional prone bed in position nursing for patients with retinal detachment who undergoing vitrectomy with intraocular tamponade .Methods One hundred and sixty-five patients who needed to take the prone position after vitrectomy with silicone oil or gas tamponade were selected and randomly divided into 3 groups:55 patients in the control group1 with the traditional U-shaped soft pillow,55 patients in the control group 2 with the hole bed and U-shaped pillow,55 patients in the experimental group with the homemade multifunctional prone bed .The daily average duration of position, the daily average sleep time,the incidence of head-bit offset, the occurrence of adverse reactions , and the recurrence of retinal detachment were observed and compared among the three groups .Results In the experimental group, the daily average time of maintaining prone position and sleeping was (19.23 ±2.36)h and (8.42 ±0.32)h, respectively, both of which were longer than those in the two control groups , the differences were statistically significant (F=69.806,75.332,respectively;P<0.01).The occurrence of adverse reactions in the experimental group were less than that in the two control groups .The recurrence rate of retinal detachment within 3 months after operation in the experimental group was 1.82%, and lower than that in the two control groups, the difference was statistically significant (χ2 =10.573,P<0.05).Conclusions The application of homemade multifunctional prone bed in patients undergoing vitrectomy with intraocular tamponade can significantly improve the patients'adherence to the prone position, effectively reduce the incidence of postoperative adverse reactions and complications , and more accurately adjust the head position , which can help to promote retinal reattachment and improve success rate of operate .

Objective To report 6 cases of hydroa vacciniforme-like cutaneous lymphoma,and to inves- tigate the relationship between this disorder and Epstein-Barr virus(EBV)infection.Methods Pathological and immunohistochemical examinations were performed in the biopsy specimens obtained from all 6 patients. Skin lesions were subjected to EBV encoded RNA(EBER)detection by in situ hybridization.Serological assay and quantification of EBV DNA were performed.Results All the 6 patients had recurrent papules, papulovesicles,necrosis and variola-like scar with chronic intermittent fever;four of the patients also presented with edema of the face,hands and feet.Pathologically,there were multilocular vesicles in the epidermis,and large numbers of infiltrating lymphocytes through the dermis.The cells were atypical with mitotic figures. Immunohistochemical staining of the lesions of 4 patients showed large quantities of cells expressing CD56, scattered cells expressing CD3 and CD45RO,and cells expressing grazyme B and T cell intracellular antigen-1 (TIA-1);a diagnosis of hydroa vacciniforme-like cutaneous NK/T lymphoma was made in these 4 cases. In the lesions of another 2 patients,the cells expressing CD3 and CD45RO,but not CD56,were observed; the diagnosis of hydroa vacciniforme-like cutaneous T-cell lymphoma was made in them.EBER was detected in the tumor cells of all the 6 patients.The IgG titers of anti-Epstein-Barr viral capsid antigen increased in all patients(1:5120 in 2 cases,1:2560 in 2 cases,1:1280 in 2 cases).The copies of EBV DNA were increased in the peripheral blood of both the two detected cases.A chronic active EBV infection was confirmed in all patients.Conclusions Hydroa vacciniforme-like cutaneous lymphoma is clinically characterized by edema of face,hands and feet,vesicular eruptions and variola like scars;histologically,it is characterized by infiltrates of atypical cells consistent with lymphoma,and necrosis in the center of vessels.NK/T is the primary immunophenotype of this disease.There is a close association between chronic active EBV infection and hydroa vacciniforme-like cutaneous lymphoma.

Objective To explore the relationship of phosphatase and tensin homology deleted on chromosome ten ( PTEN ) expression with the positioning of AMPA recepter subunit GluR2 in hippocampal neuron after stretch injury. Methods The primary rat hippocampal neuron from neonate rats were cultured and the model of stretch injury established. Western blot and TUNEL staining were used to detect PTEN expression and the positioning of AMPA receptor subunit GluR2 in rat hippocampal neuron at different time intervals after stretch injury. Results The PTEN expression was increased obviously after stretch injury, while the expressions of AMPA receptors GluR1, GluR2 and GluR3 showed no evident differences at every time point. However, AMPA receptor subunit GluR2 located at cytomembrane was down-regulated obviously after stretch injury. Conclusion The intracellular PTEN expression and the expression of AMPA receptor subunit GluR2 located at cytomembrane are on the contrary, indicating that PTEN phosphatase may participate in the transport and reintegration process of AMPA receptor subunit GluR2 located at cytomembrane.

[ ABSTRACT] AIM:To investigate the effect of silencing cell division cycle 25a ( CDC25a) gene on the prolifera-tion of human hepatoma HepG2 cells.METHODS:CDC25a gene in human hepatoma HepG2 cells was silenced by RNA interference.Real-time PCR was applied to detect the expression of CDC25a, cyclin E and CDK2 at mRNA levels in the HepG2 cells.Western blotting was applied to detect the expression of CDC25a at protein level.In addition, MTT assay, Giemsa staining and flow cytometry were used to measure the proliferation of human hepatoma HepG2 cells.RESULTS:The expression of CDC25a at mRNA and protein levels in RNA silence group was lower than those in negative control group and normal control group (P<0.05).The mRNA expression of cyclin E and CDK2 in silence group was lower than that in negative control group and normal control group (P<0.05).The cell proliferation in silence group was lower than that in negative control group and normal control group ( P<0.05) .The results of flow cytometry revealed that the cells in silence group were blocked in G1 phase.CONCLUSION:Infection of LV-CDC25a-RNAi recombinant to the HepG2 cells effec-tively inhibits the CDC25a gene expression and the proliferation of human hepatoma cells, and arrests the cells in G1 phase, suggesting that CDC25a gene may be a key target for the treatment of liver cancer.

Objective To observe clinical effects of Baixuan xiatare capsules combined with Baskin and compound Flumetasone on elderly patients with chronic eczema .Methods 160 elderly patients with chronic eczema were randomly divided into observation group and control group ,80 cases in each group , the observation group with XTR combined with Baskin , compound Flumetasone treatment , control group using Baskin combined with compound Flumetasone treatment for 4 weeks,the clinical efficacy was compared between the two groups .Results In the obser-vation group,the effective rate was 94.7%,the effective rate of control group was 82.1%,two groups have statistical significance with the efficiency difference (χ2 =5.862,P<0.05).Conclusion The XTR combined with Baskin and compound Flumetasone have better clinical effects than Baskin combined with compound flumetasone treatment with elderly patients with chronic eczema .

Objective To investigate the absorbability of the packaged infusion sets of butylphthalide,polyvinyl chloride (PVC) or polyolefin thermoplastic elastomer (TPE) infusion sets on butylphthalide for providing clinical reference in the choice of infusion sets.Methods Solution of butylphthalide was prepared as peractual concentration needed by the clinic,and dripped through PVC,TPE and packaged infusion sets respectively.The dripped solutions at different time points were collected and determined by HPLC,the contents were compared with that at 0 min.Results In the end of infusion,the adsorption rate of PVC,TPE and packaged infusion sets on butylphthalide were 0.47％,32.48％,9.62％,respectively.In addition,the absorbability of the infusion sets frown different manfacturers on butylphthalide had no significant differece.Conclusion The infusion sets of PVC had a serious absorption on butylphthalide.

OBJECTIVETo supply basis for the establishment of quality standard of Cryptolepis buchanaii.

METHODCharacters of crude drugs, microscopic characteristic as well as UV spectrum of the herb were studied.

RESULTLaticifers were found in the cortex and pith of the stem; much papillary non-glandular hair was found covering the stomata in the sub-cuticle of the leaf.

CONCLUSIONThe results can be employed as the basis for identifying the herb.

Cryptolepis ; anatomy & histology ; cytology ; Drug Contamination ; Pharmacognosy ; Plant Leaves ; anatomy & histology ; cytology ; Plant Stems ; anatomy & histology ; cytology ; Plants, Medicinal ; anatomy & histology ; cytology ; Spectrophotometry, Ultraviolet

AIM: To explore the relationship between the expression of caspase-2 and caspase-3 and the apoptosis in retinal ischemia/reperfusion (I/R) injury of rats, as well as the therapeutic effects of brain derived neurotrophic factor (BDNF)on the ischemic and reperfused retina.METHODS: This experiment was conducted at the laboratory of Affiliated Hospital of Qingdao University Medical College from February 2007 to July 2007. The models of retinal ischemia/reperfusion injury were made by transiently elevating intraocular pressure. A total of 28 rats were divided into Normal and Operative Groups. Operative group was divided into six subgroups. In each subgroup there were four rats. The left eyes of rats were used for I/R and the right eyes were used for intravitreal injection of brain-derived neurotrophic factor (BDNF) as treatment group. After reperfusion we divided our subgroups according to the reperfusion time as 1, 6, 12, 24, 48, 72 hours. The retinal ganglion cell number was counted by using optic microscope(BX-51,Olympus). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and the expression of caspase-2,caspase-3 was studied by enzyme linked immunosorbent assay (ELISA) and strept avidin-biotin complex (SABC)immunohistochemistry.RESULTS: No positive apoptotic cells were observed in the normal rats' retinae, but there were a significant number of positive apoptosis cells in 6-24 hours after transient ischemia followed by a decrease at 48 hours. The number of apoptotic cells reached a maximum at 24 hours after ischemia .The expression of caspase-2 gradually increased as early as at 6 hours, reached a peak at 24 hours, then decreased between 48 and 72 hours. Similarly, caspase-3 has the same rule with caspsae-2 in the time courses of expression in retinal tissues.BDNF administered before reperfusion inhibited the expression of apoptosis and ameliorated the retinal tissue damage. It also decreased caspase-2 and caspase-3 expression in ischemic/reperfused retina.CONCLUSION: Retinal ischemia-reperfusion can induce apoptosis of cells in the retina. BDNF rescues retinal ganglion cells (RGCs) from retinal ischemia/reperfusion injury through down-regulation of cell apoptosis and caspase-2 and caspase-3 expression. BDNF have a neuroprotective effect on retina.

AIM:To investigate the effect of recombinant lentiviral vector for RNA interference (RNAi) on the expression of fatty acid-binding protein 5 (FABP5) gene in hepatocellular carcinoma HepG2 cells and tumor formation in nude mice.METHODS:RNAi lentiviral vector was used in the experiment.Human hepatocellular carcinoma HepG2 cells were divided into 3 groups:the HepG2 cells in experimental group were transfected with the recombinant lentivirirus vector LV-shRNA-FABP5, the cells in negative control group were transfected with a control lentiviral vector LV-shRNA-NC, and the cells in normal control group were without any treatment.The nude mice were randomly divided into 3 groups.The growth of the transplanted tumor cells in the nude mice was observed.The tumor growth curve, volume and weight were de-termined 4 weeks after the cell inoculation.The expression of FABP5 was detected by real-time PCR, Western blot and im-munohistochemical staining.RESULTS:Transfection of the lentiviral vector FABP5-shRNA obviously reduced FABP5 ex-pression in the HepG2 cells.Tumor formation was all positive in the 3 groups of the nude mice inoculated with the tumor cells.Compared with normal control group and negative control group, the tumor growth slowed significantly in experimental group with smaller volume and weight.FABP5 expression in the transplanted tumor tissues was significantly down-regulated at mRNA and protein levels in experimental group as compared with normal control group and negative control group. CONCLUSION:RNAi-induced down-regulation of FABP5 effectively inhibits the growth of transplanted hepatocellular carcinoma, suggesting that FABP5 gene may be an effective target for gene therapy in treating liver cancer.