Objective To analyze the reasons of the TB outbreak in that school and to explore strategy of warning and preventing TB in schools.Methods epidemical investigation,screening close-contactors,analyzing and dealing with epidemic situation.Results 51 TB cases were detected from Oct 2003 to Mar 2005,among them,there were 42 acquired TB cases (6 sputum smear positive cases),9 tuberculosis pleurisy cases.30 of them didn't experience any health education about TB control and prevention,and know nothing about TB.Rest of them is not much better.Only tow of them is resisted by school infirmary,7 of 16 students who were diagnosed with TB before screening and took treatment were reported to local CDC.Conclusions Students lack of self-protective ability; administration of school infirmary was not well-organized,so that so many TB cases in a short period couldn't be noticed and the weakness of the report system is the major reason causing the outbreak to be long term and involving so many victims.it is emergency to enforce TB control in schools,and it is crucial to establish TB warning system in schools.

Objective:To construct I-Ad/IgG2b Fc baculovirus expression vector and express I-Ad/IgG2b Fc dimer fusion protein in Sf9 insect cells.Methods:I-Ad α,I-Ad β and IgG2b Fc gene sequences were amplified from BALB/c mouse lymphocytes by RT-PCR.I-Ad α and I-Ad β were connected with the leucine zipper sequence Fos and Jun respectively by overlapping PCR to form I-Ad α-Fos and I-Ad β-Jun.I-Ad α-Fos and IgG2b Fc fragments were ligated by restriction sites Xba I to form I-Ad α-Fos-IgG2b Fc recombination sequence.I-Ad α-Fos-IgG2b Fc and I-Ad β-Jun fragments were inserted to PPH and PP10,which were the downstream of the promoters in the plasmid pFastBacTMDual,to form pFastBacTMDual+[I-Ad/IgG2b Fc] recombinant plasmids.The constructed vector was identified by PCR,restriction endonuclease and sequencing.The recombinant plasmids pFastBacTMDual+[I-Ad/IgG2b Fc] was transferred into the DH10Bac competent cell to form recombinant baculovirus Bacmid+[I-Ad/IgG2b Fc].The recombinant baculovirus was transfected into Sf9 insect cells by liposome transfection reagent.After infected with Sf9 insect cells,the supernatant was collected and concentrated by PEG20000 to obtain I-Ad/IgG2b Fc dimer fusion protein.The fusion protein was detected by double-antibody sandwich ELISA and Western blot.Results:PCR,restriction enzyme digestion and sequencing confirmed that the recombinant vector pFastBacTMDual+[I-Ad/IgG2b Fc] had the correct sequence.The double antibody sandwich ELISA and Western blot showed that recombinant bacmid could successfully infect Sf9 insect cells,and the expressed fusion protein had the correct conformation.Conclusion:The pFastBacTMDual+[I-Ad/IgG2b Fc] baculovirus expression vector was successfully constructed and expressed in Sf9 insect cells,laying a foundation for the study of I-Ad-restricted T cells.

The therapeutic efficacy of large bone defects with traditional methods, such as autograft or allograft and biomaterial filling, is not favorable and hinders the widely clinical application. Bone tissue engineering has provided a promising way to solve this problem, and has become one of the hotspots nowadays. However, the key technology of vascularization has not been solved ideally yet. Obviously, the routine treatment has become a challenge in repairing large bone defect. In order to find a clinically applicable bone tissue engineering approach, we put forward and established a microcirculation system of osteogenesis inducted by perfusion in vivo. The "regulable osteoinduction microcirculation/microenvironment system in vivo (ROIMS)" has characteristics as follows: the culture conditions are controlled. For example, the release of a variety of growth factors can be controlled stably and quantitatively at bone defect site. Early nutrition and revascularization in the center of tissue-engineered bone can be effectively solved; some shortcomings resulted from construction of tissue-engineered bone in vivo, including secondary injury, high level technical requirements, and complex operation, can be avoided; vascularization and osteogenesis can be constructed simultaneously; simple safe and efficient operation. ROIMS perfectly combines in vitro construction and in vivo construction of tissue-engineered bone. Thereby, we can expect that this new type of technology of tissue engineered bone will be used in clinic in a near future and finally repair large segmental bone defects in clinic.

Otolaryngology

10.3969/j.issn.2095-4344.2013.25.019

Objective To study the feature of clinical and pathological type of Henoch-Sch(o)nlein purpura nephritis (HSPN) in children.Methods Clinical and pathological data of HSPN in 279 children were collected and retrospectively analyzed.Results Clinical manifestation of HSPN in 279 children with haematuria and proteinuria (107 cases,38.4％) was the most common type,followed by nephritic syndrome type (69 cases,24.7％),isolated proteinuria type (40 cases,14.3％),isolated hematuria type (29 cases,10.4 ％),acute glomerulonephritis type (21 cases,9.3 ％),rapidly progressive glomerulonephritis type (8 cases,2.9％),chronic glomerulonephritis type (5 cases,1.8％).According to the International Study of Kidney Disease in Children,the majority renal pathological type of HSPN were grade Ⅱ (133 cases,47.7％)and grade Ⅲ (109 cases,39.1％).The pathological changes of hematuria and proteinuria type were mainly grade Ⅱ (61 cases,57.0％) and grade Ⅲ (35 cases,32.7％),and the pathological change of nephritic syndrome type was grade Ⅲ (41 cases,59.4％).All of renal pathological changes,38.7％ (108/279) had codeposition of immunoglobulins A and M,30.8％ (86/279) had co-deposition of immunoglobulins A,G and M.The pathological change of nephritic syndrome type was more serious (x2 =35.989,P ＜ 0.05).Immune complex deposition was not correlated with renal pathologic classification (P ＞ 0.05).Conclusion The HSPN patients mainly show the type of hematuria and proteinuria and the type of nephritic syndrome.The majority renal pathological type of HSPN are grade Ⅱ and grade Ⅲ.In children with HSPN,the severity of the clinic symptoms is not completely consistent with the pathological changes.The pathological changes of nephritic syndrome type are more serious.To improve theprognosis of HSPN,we should make the treatment planning according to the clinical types and pathologic classification.

Objective To investigate the role of autophagy and synaptophysin (SYP) in cognitive impairment in de?pressed rats receiving electroconvulsive shock (ECS). Methods Clean and healthy adult male Sprague-Dawley rats were acclimatized to a standard laboratory environment for 7 days. The chronic unpredictable mild stress (CUMS) was used to establish the rat model of depression. Behavior tests were conducted before and after CUMS to evaluate the depression and cognition level of rats. After establishment of the model, 24 rats were randomly divided into ESC group (group E) and depression group (group D) with 12 rats in each group. The rats in group E were administered 80 mg/kg of propofol (10 mg/mL) by intraperitoneal injection, followed by ECS treatment. The rats in group D were administered propofol by intra?peritoneal injection, followed by sham-ECS treatments. The above interventions were conducted daily for 7 consecutive days. After the interventions, rats underwent behavior tests as before. Subsequently, rats were killed and specimens were collected for measurements. Immunohistochemistry was performed to examine autophagy markers such as Beclin 1 and LC3Ⅱand ELISA was used to detect SYP in the hippocampus. Results Group E after ECS significantly increased the percentage of sucrose preference (68.2%±8.7%), rearing times (7.0±1.9), total horizontal distance [(569.5±70.0) cm], es? cape latency [(21.9±5.3)s] and space exploration time [(20.5±3.9)s] compared with group D or group E before ECS. There was no significant difference in these index between groups before ECS or in group E between before and after ECS(P>0.05). Compared with group D, group E had upregulated protein expression levels of Beclin 1 and LC3Ⅱin CA1, CA3, DG as well as the area near the hippocampus and increased SYP contents (P<0.05). Conclusions Cognitive impairment in depression rats following ECS correlates with activated autophagy and increased SYP by ECS.

Objective To evaluate the changes in the expression of CXCR3 in regulatory T cells (Tregs) in the renal tissues of mice with renal ischemia-reperfusion (I/R) injury .Methods Forty-eight SPF male C57BL/6J mice ,aged 8-12 yr ,weighing 20-25 g ,were randomly divided into 3 groups ( n=16 each ) using a random number table:sham operation group (group S) ,group I/R and CD25 monoclonal antibody PC61 group (group P) . Bilateral kidneys were exposed and their pedicles were occluded for 45 min with atraumatic mini-clamp followed by 72 h reperfusion .PC61 250 μg was injected intraperitoneally at 24 h before the model was established .Blood samples were collected from the inferior vena cava at 24 and 72 h of reperfusion (T1 ,2 ) for determination of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations .Bilateral kidneys were obtained for determination of CD4+ CD25+ Foxp3+ Treg count and CXCR3+ CD4+ CD25+ Foxp3+ Treg count in renal tissues and the pathological changes of the kidney were scored .Results Compared with group S , the serum BUN and Cr concentrations and pathological scores were significantly increased at T1 ,2 in I/R and P groups ,and the number of CD4+ CD25+ Foxp3+ Treg and CXCR3+ CD4+ CD25+ Foxp3+ Treg was increased at T2 in I/R group ( P<0.05) .Compared with group I/R ,the serum BUN and Cr concentrations and pathological scores were significantly increased at T2 ,and the number of CD4+ CD25+ Foxp3+ Treg and CXCR3+ CD4+ CD25+ Foxp3+ Treg was decreased at T2 in P group ( P<0.05 ) .Conclusion Up-regulation of CXCR3 is helpful in migration of Tregs into the renal tissues of mice with renal I/R injury .

OBJECTIVE To investigate the expression of SerpinB2 and SerpinB4 genes in cells from the nasal brushings in allergic rhinitis(AR) patients, and their relationships with eosinophil numbers in nasal brushings, serum total IgE level and severity of AR. METHODS Twenty nine control subjects and 59 AR patients[29 specific IgE positive AR(sIgE-P-AR) patients, 30 specific IgE negative self reported AR(sIgE-N-SR-AR) patients] were recruited. The samples of nasal brushings and peripheral blood were collected to detect SerpinB2 and SerpinB4 gene expression and eosinophil numbers in nasal brushings, and total IgE and allergen-specific IgE level in peripheral blood. RESULTS Expression of SerpinB2 and SerpinB4 genes in cells from nasal brushings were significantly higher in sIgE-P-AR [5.17(2.33-18.96), 0.6(0.355-1.08), respectively] and sIgE-N-SR-AR [3.27(1.59-13.4), 0.75(0.42-1.64), respectively] than that in control subjects[1.21(0.1-3.285), 0.29(0.165-0.505)] (sIgE-P-AR:P=0.013; sIgE-N-SR-AR:P=0.002). Expression level of SerpinB2 and SerpinB4 had no relationships with eosinophil numbers in nasal brushings and serum total IgE level. Expression of SerpinB2 in moderate/severe AR[4.74(2.68-47.5)] was significantly higher than that in mild AR[(1.333-5.603)](P=0.025); while expression of SerpinB4 in mild AR[3.95(2.6-7.59)] was significantly higher than that in moderate/ severe AR[2.83(0.715-5)](P=0.042). CONCLUSION SerpinB2 and SerpinB4 genes might be involved in the pathogenesis of AR, and their diagnostic values in AR deserve to be evaluated with larger samples.

Objective To explore the feasibility of coelocentesis in prenatal diagnosis.Methods Coelocentesis were applied on 58 women scheduled for at 6～12 weeks of gestation,and fetal heart rate before the procedure and 1,5,30 minutes afterwards and hemorrhage of amnionic cyst was observed.Y gene of sex-determining region was detected by polymerase chain reaction,and compared with chorionic villi sample.Results The total achievement ratio of coelocentesis was 96.6% of all cases in five minutes.The coelomic fluid was successfully aspirated at first attempt in 93.2% at 7～10 week's gestation,25.0% at 6～7weeks and 50.0% at 10 ~ 12 weeks.There was no significant difference in FHR between before coelocentesis and afterwards (P＞0.05).The detection accurate rate of SRY was 91.1%,all female fetus were matched with those observation by chorionic villi sampling,and only 5 male fetus were not matched,the accurate rate was 80.8%.Conclusion Coclocentesis has the advantage of simple procedure,safe and high feasibility.The optimal time of coclocentesis is in the 7th to 10th week of gestation.