Proteolytic degradation has been a severe problem when Pichia pastoris is employed to express recombinant proteins.One alternative method to circumvent this problem is to construct protease gene disruptant.However,the main study of gene disruption is focused on nonrecombinant Pichia pastoris rather than recombinant strain.In our study,we established two different methods to directly disrupt PRC1 and KEX1 gene in recombinant Pichia pastoris.On the basis of this,we further discussed and compared the application and advantages of both methods.

OBJECTIVE To strengthen occupational safety and health(OSH) management of medical service. METHODS To analyze actuality of OSH management and internal and international laws and regulations for OSH management,and give the proposition of its construction. RESULTS On the basis of the requirement,a set of OSH management scheme was established. CONCLUSIONS It is imperative that construct occupational safety and health management laws and regulations and system in medical service.They are pacing factors for physical and mental health of medical members.

OBJECTIVE To select applicable air disinfectors that can be used at indoor dynamic state to disinfect air in hospital.METHODS Some different air disinfectors were installed different volumetric rooms.To detect indoor air bacterial content before and after running the air disinfectors at quiet state and dynamic state.To evaluate the disinfection effect of these air disinfectors according to the eliminateion rates.RESULTS At quiet state,the indoor bacteria eliminateion rate exceed 90% after the air disinfectors running 0.5h.So the disinfection were qualified.At dynamic state,these air disinfectors showed good lasting disinfection effect.CONCLUSIONS The air disinfectors solve the problem of indoor air disinfection at dynamic state.It is imperative that select and use exactly the air(disinfectors) to get good disinfection effect.

OBJECTIVE To attend to the importance and issues of using the enzyme cleaner for the reprocessing of medical instrument. METHODS The principle, usage, precautions, and the selection of enzyme cleaner were analyzed. RESULTS The cleaning of the medical instrument must use the liquid enzyme detergent that has the following characters: clear solution, no or low foam, free rinsing, flexibilities to the water temperature, and no limitation to the water quality. CONCLUSIONS For successful cleaning of the medical instrument the use of the high-quality enzyme cleaner is required. A complete cleaning of the medical instrument is the first step to assure the quality of disinfection, sterilization, and the infection control.

BackgroundReactive oxygen intermediate products lead to the oxidative injury of cells.Retinal pigment epithelial(RPE) cells produce lots of reactive oxygen intermediate products during the swallow of out disc,but how this procedure cause the persistent oxidative injury of RPE cells is poorly understood.Objective The present study was to evaluate the effect of non-lethal H2 O2 -induced persistent oxidative injury on RPE barrier in vitro.MethodsARPE-19 cell links were inoculated on 96 well plate at the density of 8×104 cells/L and the cell climbing slice of 24 well at the density of 4× 104 cells/L.The cells were cultured in DMEM/F12 medium,and the cells cultured for 24 hours in free-serum medium were used in the experiment.0-0.6 mmol/L of H2O2 were added into the medium.Cellular viability was assessed using 3- ( 4,5-dimethylthiazol-2-yl ) -5- ( 3-carboxymethoxyphenyl ) -2- ( 4-sulfophenyl ) 2H-tetrazolium(MTS) assays.Transepithelial electrical resistance (TER) was used to detect cell monolayer forming time after cultureinTrsnswellchamber.Thepermeabilityof cellmonolayer was examinedbyrhodamine isothiocyanate-dextran transepithelial flux,and immunofluorescence was used to investigate the distribution of the junction protein zonula occludens (ZO-1).ResultsThe total difference was found in the cell vitality(A490) among the different concentrations of H2 O2 ( F =991.501,P =0.000 ).Compared with 0 mmoL/L H2 O2 group,the A490 values was gradually lowed from 0.20 mmol/L H2O2 group to 0.60 mmol/L H2O2 group (P ＜ 0.05 ).H2O2 at the concentrations of ＞0.20 mmol/L lowed the viability of RPE cells.The TER value was ( 24.9 ± 1.3 ) Ω · cm2 in 11 days,( 17.8± 1.4)Ω · cm2 in 7 days after inoculation on transwell chamber,showing a significant difference between them (t=5.228,P=0.014).RPE formed the stable tight junction on day 15 with the TER value (25.9±0.9 ) Ω · cm2.The leakage amount ( relative fluorescence intensity ) of the dextran was 255.39 ± 16.44 in non-H2 O2 control group,exhibiting a significant lowing in comparison with free-cell blank group (433.08±51.53)( t =12.515,P =0.006 ),and that of H2 O2 group was significant increased in comparison with non-H2 O2 control group ( t =14.412,P=0.005).Immunofluyorescence assay showed intact intercellular ZO-1 junction in non-H2O2 control group,but the breakage of ZO-1 junction was seen in H2O2 group.ConclusionsThe results indicate that non-lethal H2O2 can destroy RPE barrier and further lead to the persistent oxidative injury of RPE cells.

OBJECTIVETo observe the effect and safety of plastering Chinese Compound Shenhuang Ointment (CSO) at Shenque (RN8) in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of qi stagnation blood stasis syndrome (QSBSS).

METHODSA prospective, multi-centered, randomized, double-blinded, controlled trial was conducted in 220 postoperative gastrointestinal dysfunction patients of QSBSS. They were randomly assigned to two groups, the CSO group (110 cases) and the placebo group (110 cases). CSO was plastered at Shenque (RN8) for 5 days after operation. The time of exhaustion, defecation, the recovery of intestinal peristalsis, integrals of TCM syndrome, and serum levels of motilin (MOT)and somatostatin (SS) were observed.

RESULTSCompared with the placebo group, the condition of exhaustion and defecation, the recovery of intestinal peristalsis on the 3rd day after operation was all improved (P < 0.05). The integrals of TCM syndrome at day 2, 3, and 4 were more significantly lowered in the CSO group than in the placebo group (P < 0.01, P < 0.05). The total effective rate of TCM syndrome was 95.3% in the CSO group, better than that in the placebo group (91.8%, P < 0.05). Compared with the placebo group, the serum MOT level increased and the serum SS level decreased at day 5 after operation in the CSO group (P < 0.05).

CONCLUSIONSThe plastering of CSO at Shenque (RN8) could advance the time of exhaustion and defecation, and improve patients' clinical symptoms. And patients could tolerate well.

Aged ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gastrointestinal Diseases ; drug therapy ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Ointments ; Postoperative Period ; Prospective Studies

Bacterial disinfectant resistance is the phenomenon that minimal inhibitory concentration or minimal bactericidal concentration of a certain disinfectant increases after a certain bacterium contacts with it many times. It exists widespread. Many species of bacteria are may resistant to a certain disinfectant, and a species of bacterium is may resistant to many disinfectant Disinfectant selectivity pressure is the extrinsic agent of bacterial disinfectant resistance. Resistance mechanisms include bacterial biochemistry structure, genetics pathway and enzymology pathway. There is relationship in disinfectant resistance and drug resistance. We should strengthen study and monitoring, enact unified standard and application specification to reduce bacterial disinfectant resistance.

Objective To report the development of clinical symptoms in a Chinese family with autosomal dominant progressive external ophthalmoplegia(adPEO).Methods Electromyologram and muscle biopsy were performed in the proband and 4 family members with the disease.Results The proband was a 57 year-old woman,who developed bilateral ptosis after the age of 30,external ophthalmoplegia after the age of 35 years old,weakness of extremities at the age of 37 years old and bulb palsy with palmus at the age of 47 years old.In the family there were 20 male and female members from five generations.All of them complained about bilateral ptosis between 26—33 years old,external ophthalmoplegia(12/15)and weakness of all extremities(14/15)between 35—45,facial and masticatory weakness(9/9)as well as dysphagia(8/9)between 44—60,accompanied with heart lesions(4/7)after 50 years old.Some patients died due to cardiac impairment.Electromyologram showed myopathic abnormalities in the examined patients. The main myopathological changes were ragged red fibers,cytochrome c oxidase negative fibers and ragged blue fibers in succinate dehydrogenase staining.Conclusions The adPEO started from extra-ocular muscles to limbs,finally facial and bulbar muscles.Heart lesions were presented in late stage and lead to death in some members.The developing process of symptoms suggested that we should pay more attention to cardiac manifestations in this disease.

Objective To investigate the axonal lesion in chronic inflammatory demyelinating polyneuropathy(CIDP).Methods Eighteen patients had undergone sural nerve biopsy.The clinical and electrophysiological distinction based on the different pathological changes were analyzed.Results Five patients with demyelination predominance which presented myelinated fiber with thin myelin.Three of them showed also mild axonal degeneration.Eight patients with axonal lesion predominance which presented Wallerian degeneration and regeneration of myelinated fibers.Three patients with mixed myelin and axon lesion of myelinated fibers and two with mild lesion.There was no significant difference between CIDP predominantly with axonal lesion and demyelination.Electrophysiological examination shows both axonal lesion and demyelination feature in some of the 2 types patients at the same time.Conclusions Axonal lesion is a common pathological change in CIDP and should not be considered as an exclusive criterion in diagnosis of the disease.Infiltration of macrophages is a common change.

Objective In this paper we analyzed the major risk factors of hemorrhagic fever with renal syndrome(HFRS) as well as its clinical manifestation,so as to provide a basis for clinical diagnosis and effective control of HFRS in Xi'an.Methods The method of retrospective study was used to collect clinical data of hospitalized patients with HFRS between 2005 and 2010 in the Hospitals for Infectious Diseases in Xi'an city,Zhouzhi and Huxian counties and then clinical manifestations of the patients with HFRS were analyzed and classified according to the national standards for clinical symptoms and test indicators of HFRS.HFRS patients matched by the ratio of 1 ∶ 2 healthy human controls were retrospectively investigated in order to obtain risk factors relevant to HFRS incidence using casecontrol study.Odds ratio(OR) method was used for single factor study.While for the multifactor study,we took the conditional Logistic regression approach.We also built models for both studies.Factor with OR ＞ 1 and P ＜ 0.05 was judged to be a risk factor.Results A total of 3090 cases information of patients with HFRS and 6018 healthy controls were collected.Of the 3090 cases of HFRS patients,sixty vaccinated patients showed atypical clinical manifestations and they were mild or moderate cases and no deaths.A total of 3030 nonvaccinated patients had obvious clinical symptoms.Severe or critical cases accounted for 39.07％(1184/3030)and 60 patients died and the fatality rate was 1.98％ (60/3030).The results of single factor Logistic regression analysis showed that of the 16 factors analyzed,the difference of 11 risk factors between the case group and the control group was statistically significant.Multivariate Logistic regression analysis showed that of the 11 factors,the difference of six factors between the case group and the control group was statistically significant.The main risk factors of suffering HFRS in the order were:exposure to rat pollutants,living in the affected areas,sitting or lying on grass fields in the affected areas,working in the affected areas,house rat infestation,and domesticated cats or dogs(OR =6.826,5.764,4.882,4.857,3.126 and 2.875; P ＜ 0.01 or P ＜ 0.05).Conclusions Vaccines are very useful in the sense that vaccinated HFRS patients tend to have mild symptoms and good prognosis.Health education in this area should be focused on the six risk factors in order to prevent the spreading of HFRS.