BACKGROUND: Basic fibroblast growth factor (bFGF), a kind of polypeptide growth factor possessing multifunctional biological activities,can protect neurons and promote the growth of nerves. It has been corfirmed that bFGF has therapeutic effects on retina ischemia/reperfusion injury (RIRI).OBJECTIVE: To establish RIRI model and analyze the effects of bFGF on cellular apoptosis of retina and the expression of regulatory gene protein.DESIGN: Randomized grouping and validating trial.SETTING: Department of Ophthalmology, the Affiliated Hospital of Medical College of Qingdao University.MATERIALS: The experiment was conducted at the Research Laboratory of Pathology, Department of Ophthalmology, Medical College of Qingdao University, from April 2002 to December 2003. Twenty-eight healthy Wistar rats were enrolled in this experiment. Four rats were randomly chosen for normal control group, the left eyes of the other 24 rats were set as normal saline control group, and the right eyes were set as bFGF group.METHODS: Normal saline control group and bFGF group adopted the rat RIRI models established by transiently elevating intraocular pressure. Normal saline of 12 μL was injected into the vitreous cavity of the left eyes of the rats in normal control group. 12 μL bFGF was injected into the vitreous cavity of the right eyes of the rats in bFGF group, 4 rats once. No administration was given in normal control group. The expression of apoptotic cells was detected and apoptosis indexes were calculated with the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method and immunohistochemical staining method at the 1st, 6th,12th, 24th,48th and 72nd hours after reperfusion and ischemia for 1 hour.MAIN OUTCOME MEASURES: ① The detection results of apoptotic cells in situ of retina tissuesat different time points after reperfusion. ②The expression of Fas and caspases-2 in retina tissues at different time points after reperfusion.RESULTS ① Comparison of apoptosis indexes of retina tissues at different time points after ischemia reperfusion: There were no apoptotic cells in the retina tissues of the rats in normal control group. As compared with those in normal saline control group, apoptosis indexes in bFGF group were significantly decreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48and 72 hours, especially at the 12th, 24th and 48th hours after reperfusion (t =5.362-5.595, P ＜ 0.05). ② The change of Fas expression at different time points after ischemia reperfusion: There was hardly any Fas expression in normal control group. As compared with that in normal saline control group, Fas expression in bFGF group was significantlydecreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48 and 72 hours, especially at the 6th, 12th and 24th hours after reperfusion (t=3.954-9.327, P ＜ 0.05). ③The changes of caspase-2 expression at different time points after ischemia reperfusion: There was no caspase-2 expression in normal control group.Compared with that in normal saline control group, the number of caspase2 positive cells in bFGF group was significantly decreased at the 6th,12th,24th, 48th and 72nd hours after ischemia for 1 hour and reperfusion (t=4.125-15.641, P ＜ 0.05).CONCLUSION: bFGF can significantly inhibit the expression of apoptosis gene Fas and caspase-2 in the ischemia and reperfusion of retina, thus reducing cellular apoptosis of ganglion cells and exerting therapeutic effects on the ischemia and reperfusion of retina.

Objective:To investigate the role of clinical pharmacists in the anti-infective treatment of one patient with complexity urinary tract infection with septic shock induced by cellulitis. Methods:Clinical pharmacists participated in one case of complexity uri-nary tract infection with septic shock induced by cellulitis, and according to the clinical curative effect and the patient' s condition change, clinical pharmacists adjusted the medication nine times and provided individualized pharmaceutical care and service in the whole process. Results:Physician accepted the suggestions of clinical pharmacists, the infection was controlled after the 38-day treat-ment, and then the patient was discharged from the hospital. Conclusion:Clinical pharmacists should participate in the clinical treat-ment, provide whole process of pharmaceutical care for severe patients and assist physician in drug treatment decisions to promote safe, effective and economical drug use.

Aim This study aimed to assess the protection of recombinant human erythropoietin (rhEPO) in light-induced injuries in human retinal pigment epithelial(RPE)cells by researching the inhibition of rhEPO for apoptosis in human RPE cells by light-induced injuries.Methods Cultured human RPE cells were exposed to light of 8 w (2 000?500) lux for 12hours,then the culture were stopped at 24 hours after 12hours light stimulation. The effect of inhibiting apoptosis of rhEPO was detected by AnnexinV-flunorescein isothiocyanate/Propidium iodium labeling and flow cytometry. The enzyme linked immunosorbant assay(ELISA)and immunocytochemical staining were used to assess the expressions of caspase-3 and Bcl-2 treated by different doses of rhEPO in light-induced injury on human RPE cells and research the protective mechanism of rhEPO by adding AG490(the special inhibitor of Jak2).Results There was a obviously increased effects on inhibiting apoptosis in every rhEPO group, which was the most conspicuous in 40 IU?ml-1 rhEPO group,and the value was (4.93?1.45)?ml-1. The decrease of expression of caspase-3 was most obvious in 40 IU?ml-1 rhEPO group, and the value was (0.125?0.029) ?g?L-1. The increase of expression of Bcl-2 was the most obvious in 40 IU?ml-1 rhEPO group and the value was 168.21?3.87. But these effects on inhibiting apoptosis in rhEPO group were restrained by adding AG490, the value of apoptosis was (11.29?2.11)?ml-1 and the density of caspase-3 increased to (0.362?0.042) ?g?L-1,the expression of Bcl-2 dropped.Conclusion It is suggested that rhEPO can inhibit the apoptosis of human RPE cells in the light-induced injuries and inhibit the expression of caspase-3 and up-regulate the expression of Bcl-2, so rhEPO can protect the light-induced injuries for human RPE cells. Its protective mechanism is accomplished principally by the pathway of combining EPO with EPOR ,then the combination activates Jak2.

AIM To investigate the effect of cisplatin on apoptosis in Scaber cell. METHODS The apoptotic cells were detected by TUNEL,HE,eletronic micrpscopy. RESULTS Treatment of Scaber cells with CDDP resulted in characteristics typical of apoptosis. CDDP induced apoptosis of Scaber cells in time and concentration dependent manner. To further investigate the mechanism of apoptosis induced by CDDP, the expressions and activity of apoptosis associated proteins such as bcl 2, bax and caspase 3 were examined using S P method.The results showed: CDDP caused time and concentration dependent decreases in bcl 2 and increased in bax proteins.CDDP bcl 2 and its translocation to perinuclei and nuclei. The expression of caspase 3 in Scaber cell were determined during apoptosis induced by CDDP. CONCLUSION Our investigetion showed that the apoptosis induced by CDDP is related to the increase of bax protein, and the decrease of bcl 2 protein. and its translocation to perinuclei and nuclei.

Objective To investigate the relationship between Livin and Ki-67 proteins, and the expres-sion and clinical significance of Livin and Ki-67 proteins in cholangiocarcinoma. Methods Fifty-five samples of cholangiocarcinoma tissue were collected in Shengjing Hospital from January 2002 to December 2003. The expres-sion of Livin and Ki-67 proteins in the 55 samples of cholangiocarcinoma tissue and 12 samples of chronic cholan-gitis tissue were detected by immunohistochemical assay. The relationship between the expression of Livin and Ki-67 proteins and the clinicopathological parameters of cholangiocarcinoma was analyzed. The results were analyzed by Spearman rank correlation coefficient, chi-square test and t test. Results The positive expression rate of Livin protein in cholangiocarcinoma was 71% (39/55), which was significantly higher than 0 (0/12)in chronic cholan-girls tissue (χ2=20.361, P<0.01). The expression of Livin protein was influenced by the differentiation of cholangiocarcinoma and the lymph node metastasis (χ2=4.193, 4.245, P <0.05). The positive expression rate of Ki-67 protein was 96% (53/55) in cholangiocarcinoma. The label index of Ki-67 in patients in clinical stage Ⅰ,Ⅱ,Ⅲ,Ⅳ,were 22%±16%, 33%±12%, 43%±15%, and 49%±10%, respectively. There was signifi-cant difference in the label index of Ki-67 between patients in clinical stage Ⅰ and those in clinical stages Ⅱ, Ⅲ,Ⅳ(t=2.307, 2.871, 3.957, P<0.05). The label index of Ki-67 was 43%±13 % in patients with local lymph node metastasis, and 34%±16% in patients without local lymph node metastasis, with statistical difference between the 2 groups (t=2.334, P<0.05). The expression of Livin protein in cholangiocarcinoma was positively correlated with the label index of Ki-67 (r=0.502, P<0.01). Conclusions Livin protein plays an important role in the pathogenesis and development of eholangiocarcinoma, and it also has correlation with the proliferating activity of cholangiocarcinoma cells. Combined detection of the expression of Livin and Ki-67 proteins may be helpful in judging the malignancy of cholangiocarcinoma and determining the prognosis of patients.

The family functioning of stroke patients is lower and related to many factors. This paper reviews the recent study in family functioning and measurement tools; the impact factors in terms of family structure, family economic status, social support and family development stage and life events.

AIM: To investigate the effect of deoxycholic acid (DCA) on the energy metabolism in human normal colon epithelial NCM460 cells.METHODS: NCM460 cells was treated with DCA at 10, 30 and 100 μmol/L for 5 d, or DCA at 100 μmol/L for 3, 5 and 7 d.After treated with DCA at 100 μmol/L for 3 d, the cells were treated with resveratrol, the activator of sirtuin 3 (SIRT3), for the next 4 d.Adenosine triphosphate (ATP) production in the mitochondria and lactate acid level were detected.The protein expression of SIRT3 was determined by Western blot.RESULTS: DCA inhibited the ATP production, increased lactate acid level, and downregulated the protein expression of SIRT3 in a dose-and time-dependent manner.Resveratrol at 10 μmol/L reversed the effects of DCA on the NCM460 cells.CONCLUSION: DCA induces the dysfunction of energy metabolism in NCM460 cells, and the mechanism may be related with SIRT3.

Hepatitis B virus (HBV) is a major cause of chronic liver disease, and frequently results in hepatitis, cirrhosis, and ultimately hepatocellular carcinoma. HBV polymerase (Pol) is an essential viral protein that is important for HBV replication and might be involved in the development of hepatocellular carcinoma. Protein-protein interactions appears to be crucial for its role. The aim of this study was to screen and identify the proteins that interact with Pol using a co-immunoprecipitation-based LC-MS/MS identification technique. The HBV Pol gene was amplified by polymerase chain reaction (PCR) and cloned into pCDNA3.1(+). The recombinant plasmid pCDNA3. 1(+)-Pol-flag was transfected into HeLa cells. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) identified 45 proteins that co-immunoprecipitated with flag-tagged HBV Pol. Eleven of these have previously been reported as proteins that interact with HBV Pol. A proof-of-concept-based Ingenuity Pathway Analysis (IPA, www.ingenuity.com) was used to characterize the functions and pathways of these 45 identified proteins and HBV Pol. Among these proteins, four proteins may play a role in three major molecular cellular networks, and are therefore worthy of further investigation.
Cell Line, Tumor ; Chromatography, Liquid ; methods ; Gene Products, pol ; chemistry ; genetics ; metabolism ; Hepatitis B ; genetics ; metabolism ; virology ; Hepatitis B virus ; chemistry ; enzymology ; genetics ; Humans ; Immunoprecipitation ; methods ; Protein Interaction Maps ; Software ; Tandem Mass Spectrometry ; methods

Objective To investigate the ability of targeting in vitro breast carcinoma and photoacoustic-mediate apoptosis of breast carcinoma of the nanoparticle probe loaded with Fe3O4 and observe its consequence of photoacoustic imaging in vitro.Methods The polymeric multifunctional nanoparticles probe that was loaded with Fe3O4 and connected with Herceptin targeting breast carcinoma was prepared in the use of double emulsion method and carbodiimide method.General physical property,the condition of Herceptin connected with nanoparticles and the ability of targeting of the probe were tested.The different concentration of nanoparticles was imaged by photoacoustic.The inhibiting effect of targeting nanoparticles on breast carcinoma cells was evaluated in vitro.Results The targeting polymeric multifunctional nanoparticles probes loaded with Fe3O,were prepared successfully with average particle diameter of (235.4± 53.75)nm and Zeta potential (-13.4 ± 4.7)mV.Fe3O4 particles dispersed on the shell of the probes.Antibody Hereeptin was successfully connected with the surface of the nanoprobes.There were massive targeting nanoprobes surround the breast carcinoma cell strains SKBR3 in the targeting group in vitro.The photoascoustic signal of the nanoprobes enhanced with the increase of Fe3O4 concentration in photoacoustic experiment in vitro.The apoptotic rate of breast carcinoma increased after the laser irradiation in the cell inhibition experiment in vitro.That proved the obvious inhibition of breast carcinoma cells was caused by photothermal effect of the prepared nanoprobes.Conclusions The prepared polymeric multifunctional nanoparticles probe that was loaded with Fe3O4 and connected with Herceptin targeting breast carcinoma can be used as a photoacoustic contrast agent,which can inhibit the proliferation of breast carcinoma by targeting photoacoustic therapy.

Aim To investigate the effects of taxol on apoptosis in Hela cell and the mechanism of apoptosis. The apoptotic cells were detected by TUNEL, HE, eletronic micrpscopy and flow cytometry method. The expressions and activity of apoptosis associated proteins such as PCNA and caspase-3 were examined using S-P and enzyme histochemistric method.The results followed as: HeLa cells exposed to Taxol undergo cell death, presenting morphological and biochemical characteristics typical of apoptosis and the apoptotic cells increased with time and concentration. In contrast to untreated Hela cells, which express low PCNA, Ones treated with Taxol expressed high amounts of PCNA. Conclusion Taxol may induced apoptosis in Hela cell. The apoptosis induced by taxol is related to the increase of PCNA protein and activity of caspase-3.