Objective To investigate the application value of amniotic cell culture and karyotype analysis in prenatal diagnosis . Methods 1 016 pregnant women in second trimester were subject to amniocentesis under the guidance of B-type ultrasonic inspec-tion ,and the cell culture and karyotype analysis were performed on the amniotic fluid which had been drawn out .Results Among 1 016 pregnant women ,1 011(99 .5% ) succeeded in the first operation of amniocentesis and cell culture .The detection rate of fetal chromosomal abnormal karyotype was 10 .3% (105/1 016) ,in which 8 .3% (85/1 016) of structural abnormality and 2 .0%(20/1 016) of quantity abnormality .856 cases were received follow-up .Conclusion Amniotic fluid cell culture and karyotype analy-sis is a safe and reliable method for prenatal diagnosis .

Objective To study the kinetic expression level of chemokines (RANTESF) in the murine infection of vulvovaginal can-didiasis (VVC), and explore the function of chemokines in local immunity of VVC. Method Sixty-three female Kunming mice, at 8 ～ 10 weeks of age, were used in this study. All animals were divided into three groups. The content variation of RANTESF in blood and yaginal fluids and CFU of vaginal fluid in each separate group of mice were detected at days 2, 7, and 14 after infection. The first group was control group. The second group was infected only one time and the third group was infected twice. The results were analyzed with SPSS 13.0 statis-tical software. Results The content variation of RANTESF and CFU in vaginal fluid reached highest at days 7 in both the first and the sec-ond groups, as well as in the blood. There were no notable changes at days 2 and 14. The content variation in vaginal fluid or blood of the second group was higher than that in the first group after infection. Conclusion CMI, as a host defense mechanism, plays an important role in protecting against vulvovaginal candidiasis, especially in secondary infection. Local innate immunity is more important than systemic in-nate immunity for protection against vulvovaginal candidiasis. Cytokine about RANTES can promote innate immunity modulation; especially the local innate immunity modulation can promote the ehemotaxis of RANTES.

Objective To investigate the effects of sevoflurane on ca2+ transsarcolemmal influx and ca2+ release function of endoplasmic reticulum in isolated outer hair cells (OHCs) of guinea pigs and the possible mechanism by which sevofhlrane acts on cochleas.Methods The experiment was performed in 2 parts.In experiment I:twelve adult guinea pigs(8 male,4 female)weighing 180-230 g were used.OHCs were mechanically sparated after enzymatic incubation.Thirty OHCs with favorable activity were divided into 3 groups (n=10 each):group I control(C);group Ⅱ low concentration sevoflurane (1.7%,group S1) and group Ⅲ high concentration sevoflurane(3.4%,group S2).The OHCs were stained with 6 umol/L Fluo-3AM in estefified form for 40 min.Group S1 and S2 were pretreated with 1.7% and 3.4% sevoflugsne respectively for 20 min.KCI 40 mmol/L was then added.The intracellular ionized Ca2+ concentration ([C2+]I) was determined byintracelhlar Ca2+ fluorescent intensity using laser scanning confocal microscope.The protocol of the experimentⅡ was the same as the experimentI.The only difference was that caffeine 20 mmol/L was added instead of KCI 40 mmol/L.Results In experiment I:there was no significant difference in baseline[ca2+]I and[ca2+]I after being exposed to sevoflurane among the 3 groups.[Ca2+]I was significanfly increased after addition of KCI as compared with the baseline[Ca2+]I and was significantly lower in group Sl and S2 than in group C and was the lowest in group S2.In experimentⅡ:the[ca2+]I was significantly increased after addition of caffeine but there was no significant difference in[Ca2+]I among the 3 groups.Conclusion Sevoflurane can inhibit voltage-dependent Ca2+ channel opening in a concentration-dependent manner but can not affect ryanodine-sensitive Ca2+ release function of endoplasmic reticuhm in isolated outer hair cells of guinea pigs.

Objective To inspect the source of an outbreak with Mycoplasma pneumoniae (Mp).Methods We carried out real-time PCR to analyze specimens collected from pediatric patients in Beijing during January 2010 to May 2012,diagnosed as pneumonia or a respiratory infection according to clinical symptoms.These positive samples were analyzed by the M-P typing system(M:multiple-locus variable-number tandem-repeat analysis,MLVA; P:P1-restriction fragment length polymorphism analysis,P1-RFLP).Results Sixty-nine specimens were tested positive to Mp by the real-time PCR in 446 specimens from pediatric patients.The infection rate was 11.69％,15.56％ and 20.00％ respectively in 2010,2011 and the first half of 2012.According to the M-P system,11 distinct genotypes were identified from 69 positive specimens,M43562P1 and M53562P1 were the two main genotypes that showed an increasing trend from 2010 to 2011,and M33562P1 and M63562P1 showed an increasing trend from 2011 to 2012 in China.Conclusion During this international Mp epidemic,the infection rate of Mp was also increase in Beijing in 2011,and M43562P1 and M53562P1 were the two main genotypes.Among them,M43562 were consistent with pop genotypes in Europe,and M53562 were consistent with pop genotype in Israel.The M-P system would be valuable to monitor the epidemic of Mp in different countries in the world.

Objective To evaluate the efficiency of using nested PCR in restriction fragment length polymorphism analysis (P1-RFLP) for genotyping Mycoplasma pneumonia (M.pneumonia) in clinical specimens.Methods Based on the gene sequence of RepMp4 and RepMp2/3 in P1 gene of reference strains M129 (type 1) and FH (type 2),two sets of inner primers were designed with a HaeⅢ restriction enzyme site (GGCC).The nested PCR was set up to detect the target DNA in clinical specimens.The amplification products were mixed and digested with Hae Ⅲ enzyme.The genotypes were analyzed by comparing with various restriction maps and the results were verified by sequencing analysis.The concentration of DNA extracted from standard and clinical strains were detected by ten-fold dilution to evaluate the sensitivity of nested PCR-P1-RFLP and P1-RFLP.M.pneumonia-positive specimens isolated from Beijing in 2012 were analyzed by the nested PCR-P1-RFLP and the results were compared with those by P1-RFLP analysis.Results The nested PCR-P1-RFLP could effectively genotype M.pneumonia in clinical specimens and the results were consistent with those by sequencing analysis.The sensitivity of new assay was 103 times higher than that of the original P1-RFLP.Of the 115 M.pneumoniae positive clinical specimens,97.4％ (112/115) were type 1 and the rest were type 2.Conclusion The nested PCR-P1-RFLP shows high efficiency for genotyping of M.pneumonia in clinical specimens.It might be useful for the surveillance of M.pneumoniae infection.

We investigated the distribution characteristics of Yersinia enterocolitica in Ningxia ,China .In accordance with the requirements of the National Yersinia enterocolitica Disease Monitoring Scheme ,Y .enterocolitica were isolated from differ‐ent kinds of specimens collected in Ningxia in 2008 to 2013 .Then they were serotyped and detected for virulence gene and ana‐lyzed the pulsed‐field gel electrophoresis (PFGE) in Chinese CDC .It was found that 173 strains were isolated from various types of 9 643 specimens ,and the detection rate was 1 .79% .There were statistical differences among detection rates in differ‐ent years and in different specimens (P<0 .01) .Pathogenic serotypes O∶3 and O∶9 carried ail gene and ystA gene were de‐tected from specimens of pigs and diarrhea patient .Non‐pathogenic serotypes O∶5 and O∶8 and non‐typeable strains didn't carry ail gene and ystA gene ,and also can't be detected from swine ,cattle ,sheep ,chickens and dogs .In conclusion ,Y .en‐terocolitica was widely distributed in Ningxia and pigs were the dominant animal host .In all pathogenic serotypes ,the highest proportion was O∶3 following by O∶9 .It was no time and regional difference in the distribution of that in Ningxia ,China .

Objective:To evaluate the effect of TBK1 overexpression on hypoxia-reoxygenation (H/R) injury in isolated mouse cardiomyocytes subjected to high glucose and the relationship with mitochondrial autophagy.Methods:Normally cultured log-phase HL-1 mouse cardiomyocytes were inoculated in a 6-well plate at a density of 1×10 6 cells/ml and were divided into 4 groups ( n=10 each) using a random number table method: control group (group C), high glucose group (group HG), high glucose and H/R group (group HG+ H/R), and TBK1 overexpression group (group TBK1). The cells were incubated in culture medium with 1% fetal bovine serum and 1% double antibody for 24 h when the cell density reached 50%.When the cell density reached 80%, pcDNA3.1 (+ ) was used as a vector to achieve TBK1 overexpression.The cells were cultured with high glucose medium (33 mmol/L) for 24 h, exposed to 94% N 2+ 5% CO 2+ 1% O 2 for 24 h in an incubator at 37℃ followed by 12 h reoxygenation in an incubator containing 5% CO 2 at 37°C to establish the model of H/R injury to cardiomyocytes subjected to high glucose.After reoxygenation, CCK-8 assay was used to detect cell viability, the activity of lactic dehydrogenase (LDH) in supernatant was detected using LDH kit, mitochondrial contents were determined using Mito-Tracter green fluorescent probe, and the expression of TBK1 and mitophagy-related proteins PINK1, Parkin, LC3B and P62 was detected by Western blot. Results:Compared with group C, the cell viability was significantly decreased, the activity of LDH in supernatant was increased, mitochondrial contents were decreased, the expression of TBK1, PINK1, Parkin and LC3B was down-regulated, and the expression of P62 was up-regulated in HG group and HG+ H/R group ( P<0.05). Compared with group HG, the cell viability was significantly decreased, the activity of LDH in supernatant was increased, mitochondrial contents were decreased, the expression of TBK1, PINK1, Parkin and LC3B was down-regulated, and the expression of P62 was up-regulated in group HG+ H/R ( P<0.05). Compared with group HG+ H/R, the the cell viability was significantly increased, the activity of LDH in supernatant was decreased, mitochondrial contents were increased, the expression of TBK1, PINK1, Parkin and LC3B was up-regulated, and the expression of P62 was down-regulated in group TBK1 ( P<0.05). Conclusion:The mechanism by which TBK1 overexpression reduces the H/R injury is related to restoring mitophagy in isolated mouse cardiomyocytes subjected to high glucose.

Objective:To observe and compare the changes of pulmonary function in patients with pulmonary tuberculosis regular treatment for 3 months.Methods:From April 2018 to June 2019, 500 tuberculosis patients who received regular anti tuberculosis treatment in our hospital were selected.The pulmonary function of patients with pulmonary tuberculosis was measured before treatment and at the end of three months; the results of pulmonary ventilation function, lung volume, diffusing capacity, and the value of forced vital capacity (FVC), maximum expiratory volume in 1 second (FEV 1), maximum expiratory volume in 1 second/forced vital capacity (FEV 1/FVC), total lung volume (TLC), residual volume (RV), carbon monoxide diffusing capacity (D LCO) were compared. Results:252 patients with pulmonary tuberculosis were included. Before treatment and at the end of three months, the abnormal pulmonary function results were 204 cases (80.95%) and 193 cases (76.59%), respectively, and the difference was not statistically significant ( P>0.05). Among them, abnormal pulmonary ventilation function is the most common, especially with obstructive, followed by abnormal diffusing capacity. At the end of three months, the proportions of patients with normal pulmonary ventilation function and normal lung volume were higher than that before treatment ( P<0.05), but there was no significant difference in the proportion of normal diffusing capacity before and after treatment ( P>0.05). The values of FVC, FEV 1, TLC and D LCO at the end of three months were higher than those before treatment, and the difference was statistically significant ( t=-6.414, -6.754, -3.863, -3.311, all P<0.01). Conclusions:Most patients with pulmonary tuberculosis have abnormal pulmonary function. At the end of the three months treatment, the normal rates of the pulmonary ventilation function and lung volume as well as the values of FVC, FEV 1, TLC and D LCO in patients with pulmonary tuberculosis were significantly improved compared with those before treatment.

Objective:To analyze the factors affecting the disappearance time of airway necrosis and repair time of airway scar stenosis in patients with ulceration necrosis tracheobronchial tuberculosis (TBTB Ⅱ) after standardized chemotherapy and bronchoscopic intervention.Methods:The clinical data of 222 TBTB Ⅱ patients admitted to Hunan Chest Hospital from January 2015 to December 2018 were collected, bronchoscopic interventional treatment was performed on time. The texture, blockage of lumen, granulation proliferation, airway stenosis of TBTB patients before treatment, the disappearance time of airway dead objects, scar repair time and stenosis degree after treatment were followed up. The disappearance time of airway necrosis and repair time of airway scar stenosis and its influencing factors were recorded and analyzed.Results:In 222 patients, 508 ulceration necrosis airway lesions were found under bronchoscopy, with a median of 2(1-6); 170(76.6%) cases of airway lesions had different degrees of stenosis before treatment. 79(35.6%) patients had tough necrosis, and 86(38.7%) patients had necrosis blocking the lumen; 132(59.5%) patients had granulomatosis. The disappearance time of airway necrosis after treatment was 1 to 32 weeks, and M( Q1, Q3) was 6(3, 9) weeks; the repair time of airway scar stenosis was 2 to 73 weeks, and M( Q1, Q3) was 14(10, 19) weeks; after treatment, there were 90.5%(201/222) patients with different degrees of scarring in the airways. Cox multiple analysis showed that the risk factor for the disappearance time of airway necrosis was tough tough necrosis ( HR=1.52, 95% CI: 1.10-2.10); the risk factor for the repair time of airway scar stenosis was the disappearance time of airway necrosis 6-9 weeks ( HR=2.73, 95% CI: 1.84-4.05). Conclusions:90.5% of patients with type Ⅱ TBTB developed airway scar stenosis after treatment. The median time for the disappearance of airway necrosis was 6 weeks, and the median time for the repair time of airway scar stenosis was 14 weeks. In the interventional process, attention should be paid to the removal of tough necrosis and the efficiency of necrosis removal to reduce the risk of airway scar stenosis.

BACKGROUND:Studies have shown that both Panax notoginseng saponins and concentrated growth factor can promote fracture healing,but there are few studies addressing their combined effects on fracture healing.Panax notoginseng saponins may accelerate fracture healing by promoting the release of concentrated growth factor-related factors over a certain period of time. OBJECTIVE:To study the effect of Panax notoginseng saponins on concentrated growth factor release and fracture healing in rats. METHODS:Eighteen 8-week-old Sprague-Dawley rats were numbered and randomly divided into three groups:Panax notoginseng saponins group,model control group and blank group.Panax notoginseng saponins group was fed with Panax notoginseng saponins for 2 weeks.Model control group was given 2 mL of normal saline for 2 weeks and blank group was fed normally.Concentrated growth factor was obtained by the centrifugation method both from the Panax notoginseng saponins group and model control group.After 1 week of normal feeding,all animals underwent modeling for femoral fracture.The Panax notoginseng saponins group and the model control group were implanted with autologous concentrated growth factor,and then the release concentration of growth factors at different time points(1 hour,1,3,5,7,9 and 11 days)were measured by ELISA.Fracture healing was assessed based on postoperative X-ray and hematoxylin-eosin staining of bone tissues. RESULTS AND CONCLUSION:Compared with the model control group,the Panax notoginseng saponins group had higher release concentrations of vascular endothelial growth factor A and transforming growth factor β at 7,9,and 11 days,Platelet-derived growth factor BB at 5,9,and 11 days,and basic fibroblast growth factor at 1-11 days(P<0.01).X-ray examinations indicated that fracture healing in the Panax notoginseng saponins group was better than that in the model control group,and fracture healing in these two groups was better than that in the blank group at 2 months after surgery.Hematoxylin-eosin staining results found that the constituent osteocyte density in the Panax notoginseng saponins group was greater than that in the model control group,and the constituent osteocyte density in these two groups was better than that in the blank group.These findings indicate that Panax notoginseng saponins can increase the concentration of concentrated growth factor-related factors.After intervention with Panax notoginseng saponins,concentrated growth factors are more advantageous in promoting fracture healing in rats.