Objective To investigate the role of oxidative stress in the epithelial-to-mesenchymal transdifferentiation (EMT) of peritoneal mesothelial cells in rat model of peritoneal fibrosis and the effect of probucol on peritoneal fibrosis. Methods The rat model of peritoneal fibrosis was induced by 4.25% high glucose peritoneal dialysis fluid (PDF). The rats were randomly divided into 4 groups:the control group, the saline group, the peritoneal fibrosis group, and the probucol group. A 4 hour peritoneal equilibration test (PET) was performed 4 weeks later. The peritoneal function and net ultrafiltration (UF) volume were determined. The level of malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in peritoneal tissue were examined. The histology of peritoneal membrane was evaluated by light microscopy. E-cadherin and α-smooth muscle actin (α-SMA) protein expression was evaluated by immunohistochemical method and Western blot.Results The mesothelial cells were detached from peritoneal membrane in peritoneal firbosis rats. Comparing with the control rats, the thickness of visceral peritoneum, the level of MDA, and the-SMA protein expression were increased while the net ultrafiltration volume, the level of GSH-Px and E-cadherin protein expression were decreased in peritoneal firbosis rats. All these changes were reversed in the rats treated with probucol.Conclusion Oxidative stress plays an important role in transdifferentiation of peritoneal mesothelial cell in the peritoneal fibrosis rats. Probucol can improve structure and function of peritoneum, and partially reverse the EMT by reducing the oxidative stress.

Objective To explore effects of fosinopril and losartan on renal Klotho expression and oxidative stress in spontaneously hypertensive rats (SHR) and the mechanisms underlying the protection against renal damage. Methods Fifteen male SHRs (22 weeks old) were randomly divided into 3 groups (n=5 in each group): a SHR group, a fosinopril group ［10 mg/(kg?d)］, and a losartan group ［50 mg/(kg?d)］. Age-matched Wistar-Kyoto (WKY) rats were chosen for a control group. Eight weeks later, tail arterial pressure, 24 hours urinary protein (Upro)，urinary N-acetyl-β-D-glucosaminidase (NAGase) were measured. Renal pathological changes were examined under light microscopy by HE staining. The renal mRNA and protein expression of Klotho were determined by RT-PCR, immunohistochemical staining or Western blot. The levels of total antioxidant capacity (TAOC), malondialdehyde (MDA), Cu/Zn superoxide dismutase (Cu/Zn-SOD), Mn superoxide dismutase (Mn-SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) were determined.Results The typical pathological characteristics of hypertensive renal damage were observed in the kidney of the SHR group．Compared with the SHR group, the systolic pressure, Upro, and urinary NAGase, the content of MDA and renal pathological damage was reduced while the renal Klotho expression and activities of TAOC, Cu/Zn-SOD, CAT, and GSH-Px were increased (P<0.05 or P<0.01) in the fosinopril or losartan group. There was no significant difference in renal Mn-SOD level among the 4 groups (P>0.05). Conclusion Fosinopril and losartan can exert protection against hypertensive renal damage through upregulating Klotho expression as well as reducing oxidative stress.

Objective To investigate the protective effects of insulin like growth factor 1(IGF-1) on cortical neurons under condition of hypoxia and the possible mechanism. Methods Cerebral cortical neurons from newborn rats were cultured under the condition of oxygen and glucose deprivation (OGD) . On day 7, neurons were treated with IGF-1 or IGF-1 plus LY294002 or PD98059 under condition of OGD or normal condition. MTT assay was used to analyze the viability of neurons in each group. The expression of total Akt and p-Akt were analyzed by Western blot. Results Compared with the control, the neuron viability was significantly higher in IGF-1 treated group under normal or OGD condition (P＜0.05). The protective effects of IGF-1 were attenuated in the presence of LY294002 but not PD98059. The result of Western blot showed IGF-1 upregulated the expression of p-Akt, which was inhibited by LY294002. Conclusion PI3K pathway may play an important role in neuroprotection afforded by IGF-1.

Objective To explore the effect of total flavonoids of Epimedium sagittatum (TFE) on the mRNA expressions of the estrogen receptor alpha (ERα) and estrogen receptor beta(ERβ) in hippocampus and hypothalamus in ovariectomized (OVX) Sprague-Dawley(SD) rats, and the mechanism of TFE against postmenopausal osteoporosis. Methods Forty-eight female SD rats,aged 10-11 months old, were randomly divided into 4 groups: a sham group, an ovariectomy group (rats were bilaterally ovariectomized), a TFE group, and a 17β-estradiol group (rats were fed with TFE and 17β-estradiol for 4 months, respectively). The RT-PCR was performed to determine the mRNA expression of ERα and ERβ in hypothalamus and hippocampus. Results Serum estradiol level, bone mineral density (BMD) of vertebra,wet weight of uterus, and the mRNA expressions of ERα and ERβ in hypothalamus and hippocampus were markedly decreased in OVX rats, all of which were reversed by 17β-estradiol treatment except the mRNA expression of ERβ. Similar results were achieved by TFE treatment except the wet weight of uterus. Conclusion TFE can improve BMD of vertebra in the OVX rats without side effects on the uterus. The mechanism may be related to increasing the mRNA expressions of ERα and ERβ in hypothalamus and hippocampus.

Objective To explore the relationship between dysfunctional beliefs and attitudes on sleep and sleep disturbances before and after a short-term pharmacotherapy in patients with major depression. Methods Sixty-six (29 male, 37 female, 34.0±9.5 years old) patients with major depression were recruited before the treatment. Dysfunctional Beliefs and Attitudes about Sleep (DBAS) questionnaire, Pittsburgh Sleep Quality Index (PSQI), Hamilton Depression Rating Scale (HDRS), and Hamilton Anxiety Scale (HAMA) were assessed before and after a 4-week antidepressant treatment. Results After the 4-week antidepressant treatment, depressive and anxiety symptoms were alleviated significantly, whereas sleep disturbance still persisted. And sleep quality was closely related with problematic sleep beliefs after the treatment.Conclusion Some evidences are provided for cognitive behavioral therapy during antidepressant therapy, and the therapy should be matched with problematic beliefs and attitudes.

Objective To prepare the infected Oncomelania hupensis by artificial method for the research on the activity, vaccine, and genetic variation of Schistosoma Japonicum (S. Japonicum).Methods The mature eggs of S. Japonicum were collected by Nylon silk method and the miracidia were incubated under appropriate conditions. Negative snails were infected with miracidia in different proportion by means of individual or collective infection to seek the best method and proportion of infection between miracidia and snails. Infected snails were divided into 12 groups in total. Ⅰ-Ⅵ groups were for individual infection and Ⅶ-Ⅻ groups were for collective infection. There were 200 snails in each group. The infection ratios between snails and miracidia in Group Ⅰ-Ⅵ or screened, numbered, and reared singly. The amount of cercariae was calculated once every 10 days until the infected snails died. Then cercariae shedding quantity, infection quantity, and mortality of infected snails in every group were compared to find the best infection method and the best infection proportion between miracidia and snails. The cercariae were collected from the first generation of infected snails and were used to infect experimental animals. The mature eggs of S. Japonicum were saved from the infected experimental animals and incubated to get miracidia. The snails were artificially infected by miracidium to get the second generation of infected snails. The developmental rates of adult worms, the egg density in fecal and liver were compared between artificially and naturally infected snails. Results In individual infection GroupⅠ-Ⅵ,the average infection value of snails were 0±0,22.7±4.2,31.7±4.5,53.0±5.3,39.3±5.9,32.7±4.7,the average fatality of snails were 21.7±3.1,25.0±3.6,31.3±4.9,44.7±6.5,78.3±9.5,89.7±13.6, and the average value of cercariae shedding from infected snails were 0.0±0.0,308.0±96.6,428.1±146.2,527.0±171.1,571.4±148.9,602.9±356.3, respectively. In collective infection Group Ⅶ-Ⅻ,the average infection value of snails were 0±0,12.3±2.5,18.7±4.7,28.3±4.2,33.3±4.7,29.3±5.5,and the average fatality of snails were 22.7±3.8,23.7±4.5,28.3±5.5,47.0±9.5,75.7±8.5,86.3±12.2, and the average value of cercariae shedding from infected snails were 0±0,244.5±57.3,292.3±74.8,347.1±100.8,477.2±142.1,447.3±161.4, respectively. The second generation of artificially infected snails was obtained successfully. The average infection rate and fatality rate for the second generation of artificially infected snails were 24.65% and 24.50%, both of which were not obviously different from that of the first generation of artificially infected snails (P>0.05). In the animal experiment, the worm growth rate for the naturally infected snails, the first or second generation of artificially infected snails were 68.50%,73.50% or 71.00%. There was no obvious difference among them (P>0.05). The fecal (or liver) eggs per gram for the naturally infected snails, the first or the second generation of artificially infected snails were 1 503±269,1 683±233, or 1 541±117 (or 6 641±1 819,6 272±1 419, or 7 263±1 643). There was no significant difference among the 3 groups (P>0.05). Conclusion Infected snails can be obtained through the artificial method by using S. Japonicum miracidia to infect snails. Individual infection has the advantage over collective infection. The optimal proportion of infection between first and the second generation of artificially infected snails in the average of cercariae shedding, infection, and fatality average of snails. There was no significant difference between artificially and naturally infected snails in the developmental rate of adult worms, fecal and liver eggs per gram.

Adeno-associated virus is a kind of DNA defective parvovirus which is non-pathogenic. Recombinant-adeno-associated virus vector comes from wild-type non-pathogenic adeno-associated virus and is highly secure, and it also has the advantages of broad host range. Recombinant-adeno-associated virus vector has become a hot spot for gene therapy and is widely used in gene therapy for cardiovascular diseases, especially for hypertension, heart failure, arteriosclerosis, and myocardial infarction.

Objective To investigate the current knowledge and attitudes towards people living with HIV/AIDS (PWHAs) in Chinese registered nurses (RNs) and describe the relationships between the nurses' HIV/AIDS related knowledge and attitudes towards PWHAs. Methods A cohort of 1350 RNs from 51 comprehensive hospitals in Hunan, China were studied over a 4-month period. A 3-stage random sampling method was used. Results The total correct rate in AIDS Knowledge Scale was 63.2%. Most nurses were good at conceptions of routes of AIDS infection and some basic characteristics, with more than 80% of the correct responses rate of relevant items. Their weakness was in the knowledge of some activities which would not transfer AIDS, such as "eating in a restaurant where the cook has AIDS may infect HIV", with less than 50% of the correct response rate of relevant items. As for attitudes, 94% of the nurses sympathized with HIV patients. About 82.7% of the nurses showed little sympathy to patients getting HIV by sexual promiscuity. Among all the AIDS related knowledge, nurses' conception of non-infectious activities was significantly related to their attitudes to HIV/AIDS.Conclusion Chinese nurses waster well about HIV/AIDS basic characteristics and the routes of infection, and most nurses sympathize with PWHAs. Their weakness is in the knowledge of non-HIV-infectious activities and they hold different attitudes to those patients getting HIV/AIDS in different ways. There are some barriers for Chinese nurses to take care of all patients equally. Professional development programs are urgently needed to remedy this situation including clarifying the nurses' misconceptions on AIDS related knowledge, developing non-judgmental professional attitudes, and using universal prevention measures when they take care of all patients.

Objective To construct human apolipoprotein O (apolipoprotein O, ApoO) expression vector and obtain recombinant fusion protein thioredoxin (Trx)-ApoO by pET prokaryotic expression system. Methods The ApoO gene fragment from the human liver cDNA library was amplified by PCR. The resulting product was cloned into pET-32a(+) vector and sequenced. The confirmed cDNA was cloned into plasmid E.coli DH10B and then transformed into E.coli BL 21 (DE3) where it was induced to express protein by isopropyl β-D-1-thiogalactopyranoside (IPTG).The fusion protein was purified by Ni-NTA resin. Results The ApoO gene was cloned by PCR and a 519 bp DNA fragment was shown on the agarose electrophoresis. The cloned gene was sequenced and demonstrated to have the same sequence as that of human ApoO gene in GenBank which justified a successful construction of recombinant plasmid. ApoO cDNA gene fragment was induced by IPTG, and a 34 kD recombinant fusion protein Trx-ApoO was tested on sodium dodecyl sulfate polyacrylamide (SDS-PAGE). Conclusion Human ApoO gene is successfully cloned and its recombinant fusion protein Trx-ApoO is expressed.

Objective To study the activation of Janus protein tyrosine kinase (JAK)/signal transducer and activator of transcription 1 (STAT1) signaling pathway and its inhibitor-signal transducer and activator of transcription-1(SOCS-1) in patients with systemic lupus erythematosus. Methods A total of 45 patients with active systemic lupus erythematosus (SLE) and 30 healthy controls were randomly selected. Western blot was performed to measure the expression of Stat1 protein and phospho-Stat1 protein (an activated form of Stat1 protein) in the monocytes after stimulation with recombinant high mobility group box1 (rHMGB1) at various time points. Expression of Stat1 protein in the skin or lesional skin was also detected. Phasic expressions of SOCS-1 mRNA in the monocytes after rHMGB1 stimulation were detected by real-time reverse transcription-polymerase chain reaction. SOCS-1 gene expression in the skin or lesional skin was also detected. Results The expression level of Stat1 proteins in the monocytes from patients with SLE was higher than that from healthy controls (t=9.16,P＜0.01) and positively correlated with SLE disease activity index (SLEDAI) (r=0.59,P＜0.01). Expression of phospho-Stat1 in the monocytes from SLE patients was time-dependently upregulated after stimulation with rHMGB1 at various time points, while expression of SOCS-1 mRNA remained unchanged(all P＞0.05). Expressions of phospho-Stat1 protein and SOCS-1 mRNA in the monocytes from healthy controls were increased transiently after stimulation with rHMGB1(all P＜0.05). Both expressions of phospho-Stat1 protein and SOCS-1 gene in the lesional skin from patients with SLE were upregulated compared with those in normal skin from healthy controls (all P＜0.01). Conclusion There are hyperactivation of JAK-STAT1 signaling pathway and negative feedback down-regulation of SOCS-1 in patients with systemic lupus erythematosus. HMGB-1 may be partly involved in the pathogenesis of SLE by the abnormal mediating function of JAK-STAT1 signal transduction pathway.