Cellulase was produced by growing Penicillium sp. NXP25 in liquid medium consisted of 5% com cob powder, 3% wheat bran, 0.35% nitrogen source No 10 and 0.3% calcium chloride. The optimum culture conditions were initial pH 5.0, 10% mycelial inoculum, temperature 29℃, shaking speed 280r/min and cultivation time 72h. When determining enzyme activity at 50℃, endo-1, 4-?-glucanase activity, extro-1, 4-?-glucanase activity, ?-glucosidase activity and filter paper enzyme activity of the supernatant of the culture were 841u/mL, 13u/mL, 24u/mL and 46u/mL, respectively. The optimum pH and temperature for the action of the above enzymes were pH 4.8 and60℃, pH5.0 and 50℃, pH 4.5 and 70℃, pH 5.0 and 55℃, respectively. Stable pH range of the above enzymes were 3.0-7.0, 4.0~6.0, 4.0~7.0 and 4.0~6.0, respectively. After incubating the enzyme complex at 65C for 30min, 24% of endo-1, 4-?-glucanase activity, 7% of extro-1, 4-?-glucanase activity, 89%of ?-glucosidase activity and 8% of filter paper enzyme activity were remained, respctively.

Adjuvants, Immunologic ; chemistry ; pharmacology ; Anti-Inflammatory Agents, Non-Steroidal ; chemistry ; pharmacology ; Clerodendrum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Phytotherapy ; Urinary Tract Infections ; drug therapy

Objective To evaluate the diagnostic accuracy of ultrasonography (US) and CT for thyroid masses. Methods Seventy-one patients with thyroid masses (13 with malignant and 58 with benign tumors) confirmed by operation and pathology were collected. The apprearances of CT and US before operation were analyzed. The apperanecs of CT and US, including the clean edge of masses, calcification and cystic degeneration necrosis were compared with those of pathologic findings. Results The numbers, cystic changes, configuration, verge, calcify and enlarge cervical lymph nodes of thyroid lesions had statistical difference in CT and US (P＜0.05). There was also difference in the internal echo, ring of halo on US and the sign of halo on plane CT (P＜0.05). The edge of thyroid mass could be displayed more clearly with US than CT (P＜0.05), however, it was similar with CT in the display of calcify and cystic changes (P＜0.05). Conclusion Both CT and US can display thyroid mass clearly. Combing of CT and US could improve the accuracy rate of diagnosis.

Objective To recognize CT signs of parapharyngeal space (PPS) changing in nasopharyngeal carcinoma (NPC) further.Methods 30 patients with proven NPC by biopsy were performed CT and MR scanning before radiotherapy. Results According to CT findings,four types of PPS display in 30 cases were found,each of which was observed and compared with MR findings and were as follows: (1)PPS ,which had no changing on CT, showed fat-filled triangular space continuously on MRI. (2) PPS, which became narrow and linear hypodense on CT,showed that fat intensity was pressed, but continuously on MRI; (3) PPS, which became narrow and linear hyperdense on CT,showed fat intensity,was pressed exceedingly but continuously either on MRI. (4)PPS,which was diminished on CT, showed the mass occupied PPS and lateral pterygoid muscles was pushed, medial fatty space of which was diminished, but intensity of medial or lateral pterygoid muscles was normal or abnormal on MRI. Conclusion (1)When PPS becomes narrow on CT,whether PPS shows hyperdense or hypodense,the mass makes a breakthrough PPS. (2) When PPS is diminished completely on CT,the mass steps PPS to the infratemporal fossa.

Objective:To observe the effect of low-intensity pulsed ultrasound (LIPUS) at different intensities on the expression of adiponectin and its receptors in C2C12 myoblasts, and to explore the potential mechanism by which LIPUS promotes the differentiation of C2C12 myoblasts.Methods:C2C12 myoblasts cultured in vitro were randomly divided into a control group and U 0.1, U 0.3 and U 0.5 groups. The control group received sham LIPUS exposure, while the U 0.1, U 0.3 and U 0.5 groups were exposed to LIPUS at intensities of 0.1W/cm 2, 0.3W/cm 2 or 0.5W/cm 2 respectively, and 1MHz for 5 min daily for 5 days. Cell viability was measured using CCK-8 assays. Fluorescence quantitative reverse transcription-polymerase chain reactions were used to detect the mRNA expression of adiponectin, adiponectin receptor 1 (adipoR1) and T-cadherin in the cells. Western blotting was employed to assess the protein expression of adiponectin, adipoR1, T-cadherin, adenosine monophosphate activated protein kinase (AMPK), activated phosphorylated adenylate-activated protein kinase (P-AMPK), embryonic myosin heavy chain (eMHC) and myogenin (MYOG). The differentiation ability of the 4 groups was measured using cell immunofluorescence chemistry. Results:After the intervention the cell viability in the U 0.1, U 0.3 and U 0.5 groups was significantly higher than in the control group. Compared with the control group, the average mRNA expression of adiponectin and the receptors of adipoR1 and T-cadherin were up-regulated significantly in the U 0.3 and U 0.5 groups. The average adiponectin, adipoR1 and T-cadherin protein expressions, and the AMPK phosphorylation level in the U 0.3 and U 0.5 groups had increased significantly compared with the control group, but all were significantly lower than in the U 0.3 group. The average protein expression of eMHC and MYOG, and the C2C12 myoblast fusion indices of the U 0.3 and U 0.5 groups were significantly higher the control group′s averages. Conclusions:LIPUS can promote the differentiation of C2C12 myoblasts. It is most effective at 0.3W/cm 2, administered for 5min/d at 1MHz with a 20% duty cycle. Its regulatory mechanism may be related to up-regulation of the expression of adiponectin, the adipoR1 and T-cadherin receptors, and the activation of AMPK phosphorylation in C2C12 myoblasts.

Exercise Test ; Female ; Heart Sounds ; physiology ; Humans ; Male ; Phonocardiography ; methods ; Pregnancy ; Stress, Physiological ; physiology ; Young Adult

Objective To study the state of erythrocyte immune function in neonates with hyperbilirubinemia,and analyze the influence of various clinical status on erythrocyte immune function.Methods Fifty-two neonates with hyperbilirubinemia were enrolled and 104 healthy neonates as the control group.The adherence rate of complement 3b-receptor on the surface of red blood cell(RBC-C3bRR) and the immune complex adherence rate of red blood cell(RBC-ICR) were detected with erythrocyte saccha-romycete rosettet test.Results 1.The level of RBC-C3bRR in neonates with hyperbilirubinemia was lower than that in control group,and the level of RBC-ICR in neonates with hyperbilirubinemia was higher than that of control group(Pa0.05).3.Comparing the neonates with unconjugated bilirubin of different concentrations,there were significant difference in RBC-ICR(Pa0.05).4.There were positive correlation between RBC-ICR and bilirubin,unconjugated bilirubin in the neonates(Pa0.05).Conclusion Erythrocyte immune function in neonates with hyperbilirubinemia is obviously lower than that of control group and it is influenced by the concentratron of bilirubin and the time of phototherapy.

Objective To learn the prevalence and trends of Brucella disease in Ningxia, in order to provide scientific basis for effective control of the disease. Methods Data of Brucella cases reported through city network from 2004 to 2009 in Yinchuan city, Shizuishan city, Wuzhong city, Guyuan city and Zhongwei city of Ningxia were reviewed and retrospectively analyzed. The data included demographic characteristics, treatment conditions and medical history so on related information. Analytical indicators include reported incidence;with patients' gender, age, regional distribution, urban and rural distribution;become chronic and associated factors;distribution of the cases reporting unit and so on. Results From 2004 to 2009, Ningxia had reported 349 cases of Brucellosis, no deaths, the annual incidence rates reported were 0.017/10 million, 0.543/10 million, 0.151/101 (295/54);The proportion of 34- to 40-year-old age group was higher than other age groups(27.5%, 96/349);Occupational distribution of patients was mainly farmers and herdsmen(70.2% ,245/349), in regional distribution of the patients, the highest percentage was Wuzhong city(61.9%,216/349), followed by Yinchuan city(22.9%,80/349);The proportion of patients in rural areas(97.4% ,340/349) was higher than urban(2.6% ,9/349);the proportion of patients converted to chronic was 11.2% (39/349). With age, the chance of patients converted to chronic was in a decreasing trend(odds ratio was 0.966);cases reported by Centre for Disease Control and Prevention accounted for 74.8%(261/349), by hospital accounted for 25.2%(88/349). Conclusions The reported incidence of Brucellosis in Ningxia is in a rapid upward trend year by year, the patients is mainly young men, the rate of converted to chronic is higher and the ability of hospital in founding and reporting of the cases is weaker.Comprehensive measures should be taken to increase funding, strengthen monitoring, and continuously improve the level of awareness and diagnosis of medical personnel for further strengthen the prevention and control of Brucellosis.

Background Proliferative vitreoretinopathy (PVR):no blood vessels,fibrous membrane cell proliferation occurs retinal surface.The development of specific mechanisms has not been completely clarified,but the retinal pigment epithelium(RPE) and platelet-derived growth factor(PDGF) for the past few years are the research hotspot.Objective To explore the effects of hypoxia on the production of PDGF-BB and proliferation in cultured human RPE (hRPE) cells.Methods In the experimental group,10,15,20,30,40 μmol/L CoCl2 were used to mimic hypoxia environment of hRPE cells,the control group did not add CoCl2 on cultured hRPE cells.By using reverse transcriptive PCR (RT-PCR) and enzyme linked immunosorbent assay(ELISA),the expression of PDGF-BB mRNA and protein was detected,and detected proliferation of hRPE cells by MTT.The cells were divided into 6 groups,different siRNAs were transfected into PDGF-BB siRNA1 group,PDGF-BB siRNA2 group,PDGF-BB siRNA3 group (because PDGF-BB has three different siRNAs,one of which is a valid siRNA),β-actin siRNA group,unrelated sequence siRNA group,only Lip2000 was plused in the blank control group.After transfection of 4-6 hours,except for blank control group,the other five groups were added to PDGF-BB mRNA and protein and its effect on the proliferation of hRPE cells obviously CoCl2 concentrations (15 μmol/L) simulated hypoxia environment of 24 hours,the detection of PDGF-BB mRNA and protein and hRPE cell proliferation rate.Results Neither PDGF-BB mRNA or protein was found in the blank control group.The production of PDGF-BB mRNA and protein in hRPE cells cultured by different concentrations were different,with significant differences among them (F=43.737,P＜0.01;F=54.612,P＜0.05),and the expressions of PDGF-BB mRNA and protein of 15 μmol/L CoCl2 group were more than other groups,with significant differences between the two groups (all at P＜0.05).MTT showed that PDGF-BB protein and hRPE cell proliferation rate in different concentions of CoCl2 groups were different,with significant differences among them(F=95.379,P＜0.01 ; F =63.375,P＜0.05),the expression of PDGF-BB protein was more and hRPE cell proliferation rate was higher in 15 μmol/L CoCl2group than other groups,with significant differences between the two groups (all at P＜0.05),postive correlation was found between PDGF-BB protein and hRPE cell proliferation rate (r =0.994,P＜0.05).The production of PDGF-BB mRNA and protein in the different siRNA transfected groups were different,with significant differences among them (F =156.330,125.650,both at P＜0.01),and the expressions of PDGF-BB mRNA and protein of PDGF-BB siRNA2 group were more than other groups,with significant differences between them(all at P＜0.05).MTT showed that PDGF-BB protein and hRPE cell proliferation rate in different siRNA transfectedgroups were different,with significant differences among them (F =73.131,98.564,both at P＜ 0.01).The expression of PDGF-BB protein was less and hRPE cell proliferation rate was lower in PDGF-BB siRNA2 group than other groups,with significant differences between them (all at P ＜ 0.05),postive correlation was found between PDGF-BB protein and hRPE cell proliferation rate (r =0.996,P＜0.05).Conclusions Proper hypoxia can induce the expression of PDGF-BB,PDGF-BB expression can significantly promote the proliferation of hRPE cells.In the transfection targeting PDGF-BB siRNA,PDGF-BB expression is suppressed,can effectively reduce the value of hRPE cell hyperplasia.

Objective:To explore the promoting effects of IL-7 and IL-2 on CD4+CD25-T cells proliferation in vitro and construct a stable culture system in vitro for CD 4+CD25+regulatory T cells from human umbilical cord blood.To compare the inhibiting effects between induced proliferated CD 4+CD25+Tregs and naturally isolated CD 4+CD25+Tregs on PBMCs functional activity.Methods:CD4+CD25-T cells and CD4+CD25+T cells were isolated from human umbilical cord blood mononuclear cells by magnetic activated cell sorting ( MACS) system and then expanded in vitro.Four different concentration levels of IL-7 combined with proper concentration of IL-2 were added as inducer and the efficiency and optimal concentration of IL-7 on inducing,CD4+CD25-T cells were analyzed via 4 different methods.Flow cytometry method was used to detect the changes of CD 4+CD25-T cells.The inhibitory effect of expanded CD 4+CD25+T cells on peripheral blood mononuclear cells (PBMCs) was tested by MTS.The expressions of Foxp3,IL-10 and TGF-βgenes in CD4+CD25+T cells were test by RT-PCR.Results:The CD4+CD25+T cells from each groups were expanded significantly after three weeks of culture.The results indicated that use of IL-7 combined with IL-2 resulted in the highest cell expansion comparing to the other groups.It was shown by the inhibitory test that the expanded CD 4+CD25+regulatory T cells could inhibit the proliferation of PBMCs ,but IL-7 induced CD4+CD25+regulatory T cells exerted weaker suppressor activity than natural regulatory T cells .Only IL-7 (4 ng/ml) and IL-2 (2 000 U/ml) induced CD4+CD25+regulatory T cells showed the strongest killing activity.Conclusion:We successfully expand CD4+CD25+regulatory T cells in vitro.The protocol is established in which the use of mAbCD 3/CD28 combined with IL-7 and IL-2 resulted in the highest cell expansion ,and intensely expressed cell phenotype of CD 4 and CD25.