Objective To construct recombinant lentivirus silence vector aiming at rictor gene in mTORC2 specific protein, and to investigate its regulation on mTORC2/SGK1 signal pathway and the effect on pulmonary alveolar epithelial sodium ion chan-nel,as well as the role in acute respiratory distress syndrome(ARDS)and acute lung injury.Methods The interfering vector plas-mid and empty vector plasmid of target gene rictor were constructed,which and the lentivirus packaging system were co-transfected to 293T cells.The viral supernatant was collected,centrifuged,concentrated and purified for obtaining recombinant lentivirus.The virus titer was detected and the virus was infected to A549 cells.Stable cell lines were screened.RT-PCR was used to confirm the silencing situation of target gene rictor.The expression situation of various signal indexes in this pathway was detected by PCR and Western blot.Results The recombinant lentivirus of silence gene rictor was successfully constructed and transfected to A549 cell for obtaining stable cell lines.Compared with blank and control groups,the mRNA levels of rictor,downstream SGK1 andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased (P <0.05 ).Meanwhile,the protein levels of rictor,downstream SGK1,P-SGK andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased compared with the other two groups(P<0.05).Conclusion Silence rictor gene has the obvious regulation effect on mTORC2/SGK1 signal pathway,meanwhile affects the expression of pulmonary alveolar epithelial cellular α-,β-and γ-ENaC at gene and protein level.It is speculated that mTORC2/SGK1 may be an important signal pathway for regulating the clearance capacity of pulmonary alveolar epithelial cells on pulmonary alveolar fluid and simultaneously affecting the pulmonary edema formation.

It has always been controversial whether a single allergen performs better than multiple allergens in polysensitized patients during the allergen-specific immunotherapy.This study aimed to examine the clinical efficacy of single-allergen sublingual immunotherapy (SLIT) versus multi-allergen subcutaneous immunotherapy (SCIT) and to discover the change of the biomarker IL-4 after 1-year immunotherapy in polysensitized children aged 6-13 years with allergic rhinitis (AR) induced by house dust mites (HDMs).The AR polysensitized children (n=78) were randomly divided into two groups:SLIT group and SCIT group.Patients in the SLIT group sublingually received a single HDM extract and those in the SCIT group were subcutaneously given multiple-allergen extracts (HDM in combination with other clinically relevant allergen extracts).Before and 1 year after the allergen-specific immunotherapy (ASIT),the total nasal symptom scores (TNSS),total medication scores (TMS) and IL-4 levels in peripheral blood mononuclear cells (PBMCs) were compared respectively between the two groups.The results showed that the TNSS were greatly improved,and the TMS and IL-4 levels were significantly decreased after 1-year ASIT in both groups (SLIT group:P＜0.001;SCIT group:P＜0.001).There were no significant differences in any outcome measures between the two groups (for TNSS:P＞0.05;for TMS:P＞0.05;for IL-4 levels:P＞0.05).It was concluded that the clinical efficacy of single-allergen SLIT is comparable with that of multi-allergen SCIT in 6-13-year-old children with HDM-induced AR.

0.05].In 48 patients without lymph node metastasis,there was no significant difference between the OPN-positive and OPN-negative patients in survival times.Multivariate analysis revealed that survival times was associated with patients′sex(P=0.032)and TNM stage(P

Objective To investigate the clinical treatment efficacy of mutabilis coated nasal ointment to postoperative nasal septal reconstruction.Methods After surgical reconstruction,116 cases were divided into the treatment group and control group ac-cording to random method,58 cases in each group.The treatment group was given mutabilis coated nasal ointment combine with physiological seawater nasal care device;the control group was only given physiological seawater nasal care device in the same time. The effects were observed for 6 months.Results The total effective rate of treatment group was higher than the control group (98.28%),the two groups had significant difference(P <0.05).Postoperative follow-up,the bleeding rate in the treatment group have no significant difference compared with the control group(P >0.05);the incidence of infection and cavity stricture in the treat-ment group were significantly lower than the control group(P <0.05).Conclusion The application of mutabilis coated nasal oint-ment can promote the wound healing of nasal cavity,but also can prevent the operation adhesion,and shorten repair time,so as to improve the clinical effect of operation.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Humans ; Occupational Exposure ; Phthalic Acids ; analysis ; Spectrophotometry ; methods ; Workplace

Objective To investigate the effect of endoplasmic reticulum stress in hyperlipidemia rats with acute pancreatitis(AP).Methods Forty-eight SD rats were divided into AP group(n=18,in- duced by injection of 40?tg/kg caerulein in normal rats twice with 2 hrs interval),normal control(n=6), byperlipidemia group(n=6,fed with a high-fat diet for 8 weeks)and hyperlipidemia AP group(n=18,in- duced by injection of 40?g/kg caerulein in hyperlipidemia rats twice with 2 hrs interval).The rats were sacrificed at 9,12,24 hrs(6 of each),respectively.Serum level of amylase was tested and the pathologi- cal changes of the pancreatic tissues were observed.The index of pancreatic apoptosis was assessed by TUNEL method.The expression of GRP78/Bip(glucose regulated protein)protein was determined by immunochemistry,the endoplasmic reticulum stress related molecules of XBP-1 splicing(X box binding protein),CHOP/GADD153(C/EBP-homologous protein or growth arrest and DNA-damage-inducible gene 153),caspase-12 were analyzed by RT-PCR.The dynamic expressions of GRP78/Bip and caspase- 12 were determine,d by Western blot.Results Artier 8 weeks of fat diet in hyperlipidemia rats,the ser- um levels of triglyceride[(0.99?0.38)mmol/L]and cholesterol[(3.17?0.18)mmol/L] were signifi- cantly increased(P

Objective To investigate the clinical and molecular epidemiologic features of human rhinovirus (HRV) in children with acute lower respiratory tract infection (ALRTI) in Shanghai by detecting HRV from bronchoalveolar lavage fluid (BALF) .Methods One hundred and thirty-five BALF samples were collected from children with ALRTI at Children′s Hospital of Fudan University from January to June in 2014 . The specimens were tested by one-step real-time reversible transcription (RT )-polymerase chain reaction (PCR) for HRV .Gene fragments of VP4/VP2 capsid protein of the HRV genome were amplified and sequenced .The phylogenetic trees of HRV strains were drawn .Chi-square test was used for statistical analysis .Result Among these 135 samples ,6(4 .4% ) were HRV positive , including 4 positive for HRV-A and 2 positive for unknown genotype .The age of children with HRV positive was 8 months to 8 years ,including 5 cases ≤ 3 years old .The nucleotide homology within 4 HRV-A samples varied 76 .5% -100 .0% .These 4 HRV-A sequences were respectively compared with reference strains of HRV-A ,HRV-B and HRV-C in GenBank .The nucleotide homology varied 73 .1% -91 .7% ,57 .8% -66 .0% and 59 .0% -65 .0% ,respectively .Among 135 children with ALRTI ,124 were diagnosed with ordinary pneumonia including 4 (3 .2% ,4/124) HRV positive and 11 were diagnosed with severe pneumonia (with respiratory failure and/or heart failure) including 2 HRV positive .All children with HRV presented with fever and cough .Most of them (5/6) were co-infected with other pathogens .In 5 children with HRV positive ,the peripheral blood leukocyte counts were less than 10 × 109/L , neutrophil percentages were less than 0 .50 ,C-reactive protein was less than 8 mg/L and procalcitonin was less than 0 .05 μg/L .All the children were clinically cured .Conclusions The detection of HRV in BALF samples indicates that HRV is one of the pathogens in children with ALRTI in Shanghai and HRV-A is more common .Gene fragments of VP4/VP2 capsid protein of HRV pathogenic strains display a high degree of genetic variability .

?AIM: To discuss the application of anterior segment optic coherence tomography ( AS-OCT ) in the diagnosis of corneal ulcer.?METHODS: The cross linear scan was used in 88 patients ( 88 eyes ) with corneal lesion by AS-OCT to gather the image data, observe the pathological changed tissue by measuring all layers for patients with initial inspection, providing important visual images and data for treatments. All the patients were followed up for 2mo.?RESULTS:Clear images with structure of all layers were obtained. It can provide the intuitive image data and scan the same position and show the changes during the treatment.?CONCLUSION: AS-OCT can discover the important condition immediately. It also can monitor course of disease dynamically, provide the intuitive image data for clinical treatment.

Objective To express the plasminogen activator(Pla) of Yersinia pestis and one of its gene fragments,and to detect their immunological reactivity.Methods The pla gene and its specific gene fragment pla-c were amplified by PCR using the EV76 strain as a template.PCR products were then ligated with the plasmid pET32a (+).The recombinant plasmids pET32a (+)-pla and pET32a (+)-pla-c were subsequently trausformed into E.coli BL21 (DE3).The expressed products were purified by HIS affinity chromatography,and their immunological reactivity was detected by Western blotting.Results The recombinant Pla(52.8 × 103) was expressed as inclusion bodies,and the recombinant Pla-c protein (24.0 × 103) was expressed in the soluble form.These two recombinant proteins reacted with anti-Yersinia pestis EV76 rabbit sera.Conclusions The recombinant Pla and its specific fragments have displayed immunological reactivity,and can be served as an alternative diagnosis method for Yersinia pestis.

Objective To collect the data of present basic constructive situation of intensive care units (ICUs) in second and tertiary grade hospitals in Xinjiang Uygur Autonomous Region and to provide a data base beneficial to the construction of ICUs in the whole region.Methods The information including basic situation of ICUs in a total of 49 second and tertiary grade hospitals as samples in Xinjiang Uygur Autonomous Region were investigated by questionnaire survey from December 2012 to April 2013. The contents of questionnaire chiefly included the basic situations of hospitals, ICU constructions, ICU human resources and allocation of equipments.Results Forty-nine questionnaires were sent out, and all of them gave answers, the recovery rate being 100%. In the whole region of Xinjiang, there were 49 second and tertiary grade hospitals with establishment of 66 various types of ICUs. ① Medical unit basic situation: the second grade A hospitals accounted for 59.2%, and the tertiary grade A hospitals, 34.7%. There was a larger proportion of local hospitals (67.3%); most of the hospitals had beds over 500 (57.14%), the designed hospital bed number was 41 403, and 35 442 beds were open up for service (85.60%). ② The ICU basic construction situation: the construction of ICUs concentrated mainly after the year 2000, from 2000 to 2010, a total of 37 units, and from 2010 to 2012, 12 units were constructed; when they opened to the public, the beds were relatively few, and the ICUs containing less than 10 beds occupied 79.00%. In the 66 ICUs surveyed, there were 43 general ICUs, accounting the largest proportion, followed by 7 emergency ICUs, and the least were 1 neurosurgery and 1 cardiac surgery ICU. In the 66 ICUs, all together 765 beds were prepared, but actually 808 beds were opening up to public; most of ICUs had 5 - 20 beds, accounting for 71% in all the ICUs. At present, in the whole Xinjiang region, the rate of beds in ICUs in various grades of hospitals opening to the outside for use accounted for more than 80% of which 56% ICUs were almost turning round fully. ③ ICU human resources situation: totally, there were 484 doctors in ICUs in the whole region, the ratio of full-time doctor to nurses was 1:3.50, the ratio for bed to physician 1:0.55 and the ratio for bed to nurse 1:0.92. The title of doctor was primarily resident, and the various title gradient levels were good. Most doctors graduated from a regular medical college, and the doctors having obtained a master degree were few. The physician professional backgrounds were mainly critical disease medicine, emergency medicine and respiratory medicine, the professional relevance being good; the chief way of ICU physicians to engage in advanced study of their professional training was in China, and generally they joined professional continuing education programs poorly, mostly joining once a year or non at all. The physicians who obtained the identification of Chinese critical care medicine specialist accounted only for 23.56%. ④ ICU equipment allocation situation: according to the requirements of ICU construction guidelines, the equipments must be allocated including bedside monitors (703 units), ventilators (516 units) and infusion pumps (702 units), occupying the highest proportion. In the optional equipments, there were enteral nutritional pump (89 stations), blood glucose monitoring device (57 units) and anti-decubitus mattresses (54 units), the proportion being the largest, bispectral index monitor (2 units), extracorporeal membrane oxygenation (ECMO, 1 table) and gastric mucosal carbon dioxide tension pHi meter (0 unit), the proportion being the least or non at all.Conclusions In the whole Xinjiang region, the construction of ICUs is still at the developing stage, and needs to further strengthen the standardized construction. The human resources, equipments, etc. are not distributed in balance, and it is necessary to move forward the adjustment to benefit the development of ICUs in the whole region.