Objective To develop a quick quantitative detecting method for point of care testing (POCT) of human serum procalcitonin (PCT) by fluorescence immunochromatographic technology.Methods Applying a double-antibody sandwich immunofluorescent assay (one antibody coated on the nitrocellulose membrane and the other antibody labeled with fluorescent micropaticles) to develop a PCT quantitative detecting kit by immunochromatography technology.The kit was used to test PCT in 472 serum samples from suspected bacterial infection patients of Guangzhou Red Cross Hospital,including 240 male and 232 female patients.The methodology and diagnostic performance were evaluated in the aspects of linearity,precision,accuracy,specificity,stability experiments and comparison with foreign PCT detecting kits.Results The report range of the PCT quantitative diagnostic kit was 0.1-125.0 μg/L The coefficient of variation (CV)values of repeat 20 tests for low,median,and high concentration control samples respectively were all less than 15％ and bias can be acceptable (P ＞ 0.05).Common interfering substances in human serum specimens such as bilirubin (2.0 g/L),triglyceride (30.0 g/L) and cholesterol (15.0 g/L) were found no significant affect on quantitative detection of PCT.The shelf time of the PCT diagnostic kit should be longer than 12 months as the relative deviation of detected concentrations of 0.5,1.0,22.0,65.0 μg/L PCTcontrol sample can be controlled less than 20％ within 14 months.Considering VIDAS BRAHMS PCT to be the standard quantitative test for PCT,472 serum samples were detected by both our kit and the control VIDAS BRAHMS PCT kit simultaneously,which showed high correlation (YVIDAS =0.180 + 1.006Xwondfo,R2 =0.988,P ＜ 0.01) and low deviation (Z =-1.6,P ＞ 0.05) without statistic significance between two methods.And the results of these two diagnostic kits showed good consistency as the area under curve of the receiver operating characteristic (ROC) of Wondfo-PCT at the three cut-off values (0.5,2.0,10.0 μg/L)were 0.997,0.994,0.998 respectively,P ＜ 0.01,using diagnostic result of the control product as standard.Kappa values were 0.899,0.905,0.973 respectively.Conclusions The method of quantitative detection of PCT by fluorescence immunochromatography for POCT was established in this study.All the observed indicators reached the clinical diagnostic requirements and can be applied for the quick detection of clinical human serum PCT.

Objective To explore the efficacy and safety of the bronchoscopic high frequency electrocoagulation combined with balloon dilatation in treating tuberculosis inflammatory airway constriction. Methods According to the different methods of treatment, 55 patients with tuberculosis airway constriction were randomly divided into two groups, the balloon dilatation group (26 cases) and combination group (29 cases). The patients in balloon dilatation group underwent bronchoscopic balloon dilatation and the patients in combination group underwent bronchoscopic balloon dilatation combined with high frequency electrocoagulation. The patients of the two groups accepted endoscopic therapy once a week. Effective rate of recanalization for the narrow airway, frequency of effective treatment and the time of tuberculosis bacterium vanishing was recorded. Intraoperative and postoperative complications were also observed. Three months after the treatment, all patients accepted bronchoscopic to observe and assess the airway restenosis rate. Results After treatment, the effective rate in balloon dilatation group and combination group had no significant difference[69.2%(18/26) vs. 89.7% (26/29 )](P＞ 0.05 ),but frequency of effective treatment and time of tuberculosis bacterium vanishing had significant difference[(3.5 ±1.3) times vs. (1.5 ± 1.1) times, (23.3 ±3.6) d vs.(13.2 ±2.3) d](P<0.01). There was no significant difference on the intraoperative and postoperative complications between two groups (P＞0.05). The airway restenosis rate was 33.3%(6/18) in balloon dilatation group and 7.7%(2/26) in combination group after treatment for 3 months (P ＜0.05). Conclusions Combination of bronchoscopic balloon dilatation and high frequency electrocoagulation is an efficacy and safety way for the tuberculosis inflammatory airway stenosis. It can reduce the frequency of interventional therapy, shorten the time of tuberculosis bacterium vanishing, and may also decrease the airway restenosis rate.

Objective To explore the effect of Jisuikang on neural functional recovery, and expression of brain-derived neurotrophic factor (BDNF) protein and mRNA level after spinal cord injury (SCI). Methods 144 female Sprague-Dawley rats, weighted 180 to 220 g, were used for experiment. 24 rats were randomly extracted into sham group (Group A), which had their vertebral plates and spines bitten away only.The others were randomly divided into model group (Group B), prednison group (Group C), and high, middle and low doses of Jisuikang group (Groups D to F) after SCI, 24 rats in each group. Group C was given 0.06 g/(kg ⋅ d) prednison, and Groups D to F were given 50, 25 and 12.5 g/(kg ⋅d) Jisuikang respectively, which were given 20 ml/(kg ⋅d) volume by intragastric administration. Groups A and B were given the same volume of normal saline (NS). The Basso-Beattie-Bresnahan (BBB) scores and oblique board test were applied to test the postoperative results 24 hours, 3, 7 and 14 days after SCI. The rats were executed and the spinal cord tissues were extracted 3, 7 and 14 days after SCI. Immunohistochemistry, Western blotting and RQ-PCR were applied to test the expression of protein and mRNA of BDNF. Results BBB scores and angle of oblique board test were significantly lower in Groups B to F than in Group A 24 hours after SCI (P<0.01). BBB scores were higher in both Groups C and E than in Group B 3 to 14 days after SCI (P<0.05), and was significantly higher in Group E than in the other groups 14 days after SCI (P<0.01). The results of immunohistochemistry and Western blotting showed that the protein expression of BDNF were significantly higher in Groups C and E than in Group B at different time points in the injured area after SCI (P<0.01), while there was no significant difference between Groups C and E (P>0.05). The results of RQ-PCR showed that prednisone and Jisuikang promoted the expression of BDNF mRNA. Group C (prednisone) had a most obvious effect at the beginning while Group E was better than Group C 14 days after SCI. Conclusion Jisuikang can promote the neural functional recovery and the expression of BDNF on both protein and mRNA level in SCI rats.

Objective To discuss the effects of repairing nasal ala defects by free transplantation of autogenous auricle composite tissue flap. Methods 50 cases with nasal ala defects were repaired by free auricular composite tissue flap transplantation from Janu 2003 to May 2013. The defects size was 0.5cm x 0.7cm~1.3cm x 1.5cm. According to the size of the defects, full thick wedge-shaped auricle composite tissue flap were cut off, then inserted into the nasal ala defects area and fixed stablely, the donor sites were sutured directly avoiding ear cartilage. Salvianolate and hyperbaric oxygen were used in postoperative treatment for 5-7 days. Results 48 cases achieved good results, 2 cases had partial necrosis of composite tissue flap after operation, also achieved good results after reoperation. Over 3 months~5 years follow-up, all cases had satisfactory results. The volume of auricle composite tissue flap reduced less than 10%. Surgical incisions had a linear scar, good color matching, nostril symmetry. All patients were satisfied with the overall appearance. Conclusions Autogenous auricle composite tissue flap free transplantation for repairing middle and small size of nasal ala defects can recover ala formation and structure very well, nasal appearance can be improved greatly with nostril symmetry. There is no hypertrophic scar in donor sites. This method is simple and easy,and is also a good method for repairing nasal ala defects.

Objective To develop a rapid quantitative detecting assay for point-of-care testing ( POCT ) of N-terminal pro-brain natriuretic peptide ( NT-proBNP ) in serum by the fluorescence immunochromatographic technology.Methods Applying double-antibody sandwich assay to establish the quantitative NT-proBNP kit.The performance of quantitative NT-proBNP kit was evaluated by the sensitivity , specificity, accuracy, precision, stability and clinical effectiveness.It compared the research kit and conference kit by the parallel experience in the 1 056(605 males, 451 females)serum specimen collected from Guangdong Provincial People′s Hospital, Sun Yat-sen Memorial Hospital and Children′s Hospital of Zhengzhou between February 2013 to April 2014.Statistical significance of the results was assessed by correlation analysis , linear regression , receive operating characteristic ( ROC) curve analysis , negative and positive consistent.Results The report range of the NT-proBNP kit was 18-35 000 ng/L.The coefficient of variation ( CV) values for low , median and high concentration calibrators respectively were all less than 15%.Common interfering substances in human serum specimens such as bilirubin , triglyceride and cholesterol were found no significant affect on NT-proBNP antigen detection and the CV were no more than 15%.According to the results of detection for calibrators , the shelf time of the NT-proBNP diagnostic kit should be longer than 12 months.The NT-proBNP kit and reference kit had good correlation ( Y=1.048 9X developed reference +121.54, R2 =0.956 6, n=1 056) to detect the target protein through the parallel experiments and the deviation of the quantitative results of clinical serum samples showed no statistical significance (Z=0.88, P=0.379>0.05).The clinical assays of two different diagnostic kits showed good consistency based on the ROC curve evaluation which is compared by two cut-off values (300 and 450 ng/L).The areas under ROC curve were 0.981 and 0.978 respectively.Conclusions A novel NT-proBNP chromatographic quantitative immunofluorescence detection method was developed in this study .The performance evaluation data indicated that the kit is suitable for rapid detection of serum NT -proBNP.

BACKGROUND:Studies have shown that Clematis chinensis Osbeck can promote articular chondrocyte proliferation, maintain and promote the synthesis of cartilage proteoglycan and type II col agen in chondrocytes. Low-intensity pulsed ultrasound can effectively improve the carticular chondrocyte proliferation, increase the cellmembrane permeability, and promote chemicals or genes delivery, so as to improve the biological effects of chemicals.
OBJECTIVE:To observe the effect of low-intensity pulsed ultrasound mediated Clematis chinensis Osbeck on the proliferation of cartilage cells and the expression of type II col agen and transforming growth factor (TGF)-β1 of rabbit knee articular chondrocytes cultured in vitro, and explore the effect and mechanism of Clematis chinensis Osbeck mediated by low-intensity pulsed ultrasound on cartilage damage.
METHODThe rabbit knee articular chondrocytes were isolated and cultured in vitro. cells of the second generation in logarithmic growth phase were randomly divided into four groupcontrol group, low-intensity pulsed ultrasound (LIPUS) group, Clematis chinensis Osbeck (CCO) group, and CCO+LIPUS group. After cells were cultured under different conditions for 3 days, cellproliferation was measured with cellCount Kit-8 assay. The secretion of type II col agen was detected by cytochemical staining method, and the expression of TGF-β1 was assayed by western blot analysis.
RESULTS AND CONCLUSION:The number of cells in the other three groups were significantly higher than that in control group at 3 days after culture (P<0.05), while the CCO+LIPUS group had the most cells. Cytochemical staining showed that the area and absorbance value of type II col agen in cartilage cells were significantly increased in CCO+LIPUS group, compared with control group (P<0.05), and the difference with control group was larger than the amount of that of LIPUS group and CCO group with control group. Western blot analysis showed that TGF-β1 was significantly expressed in CCO+LIPUS group, and the relative gray value was significantly higher than the other groups (P<0.05). Both low-intensity pulsed ultrasound and Clematis chinensis Osbeck can promote the proliferation of cartilage cells and expression of type II col agen and t-β1 of rabbit knee articular chondrocytes cultured in vitro. The combined method could produce synergistic effects.

Objective To evaluate the growth inhibition and apoptosis of human monocytic leukemia THP-1 cell line by using 5,8-dimethyl-2-β-hydroxyisovalerylshikonin (SK36) and explore its preliminary mechanism. Methods CCK colorimetric assay and cell counting was used to examine the growth inhibition of shikonin on THP-1 cells. The apoptosis of THP-1 cells was detected by Annexin V/PI double labeling. The activation of Caspase-3 apoptosis pathway was determined by FCM. The apoptosis and the necrosis of THP-1 cells were detected by the laser scanning confocal microscopy. Results When the THP-1 cells were treated with SK36 at 1.02 μg/ml for 24 h and 48 h, the growth inhibition was dose-dependent. The cell apoptotic rate of THP-1 cells treated with 1.02 μg/ml evaluated by FCM with Annexin V/PI double labeling staining were (40.61 ±2.13) % and (67.40±9.15) % at 24 h and 48 h after treatment, respectively, which were significantly higher than that of the control group [(16.97±0.61) %] ([ = 18.444, t = 9.528, P <0.01). SK36 could induce THP-1 cells apoptosis involving the activation of Caspase-3 (F= 323.61, P<0.01). Conclusion SK36 can induce human THP-1 cells to undergo apoptosis, and its primary mechanism was to activate the Caspase-3.

Objective To understand characteristics and research status of literatures related to surgical site infec-tion(SSI)in China.Methods Literatures about SSI published between January 2000 and March 2016 were retrieved from China National Knowledge Infrastructure(CNKI),VIP database,Vanfang Database,and China Biology Medi-cine(CBM)database. Bibliometric method was adopted to analyze external and internal characteristics of literatures. Results A total of 1036 articles in Chinese were included,40(3.86% ),189(18.24% ),and 807(77.90% )were published in 2000-2005,2006-2010,and the first quarter of 2011-2016 respectively. Articles were mainly pub-lishedinChineseJournalofNosocomiology(n= 226,21.81% ),ChineseJournalofInfectionControl(n= 53, 5.12% ),andChineseJournalofDisinfection(n= 27,2.61% ). The research fields included risk factors(n= 277, 26.74% ),infection rates (n= 261,25.19% ),antimicrobial application (n= 208,20.08% ),and pathogens (n=153,14.77% );the infection rates were higher in general surgery and neurosurgery,the main pathogens were Esch-erichiacoli,Staphylococcusaureus,and Pseudomonasaeruginosa,risk factors mainly included the types of incision, duration of surgery,diabetes,age,and body mass index.Conclusion In recent years,articles about SSI research in-creases significantly,research in etiology and epidemiology has gained substantial achievement,but in the interven-tion and economics is still weak,suggesting that SSI research in economics,risk management,and behavioral aspects should be strengthened.

AIM: To investigate the changes of the L-type calcium channel subunit expression and calcium currents (I_~Ca -L) in cultured rat ventricular myocytes infected by coxsackie virus B_3 (CVB_3). METHODS: Primary cultured neonatal rat ventricular myocytes were infected with CVB_3. The changes of L-type calcium channel subunits mRNA in normal and infected myocytes were measured by semi-quantitative reverse transcription-polymerase chain reaction. I_~Ca -L was recorded in two groups respectively using whole cell patch-clamp techniques. RESULTS: The expression of ?_1 and ? subunits of L-type calcium channel mRNA increased in the infected group compared with the normal one (4.00?0.07 vs 2.21?0.41, P0.05). The average current density of I_~Ca -L significantly increased by CVB_3 infection [(-8.66?0.99) pA/pF vs (-6.97?1.75) pA/pF, P