Objective To construct quality control materials for chlamydia trachomatis (CT) polymerase chain reaction detection and evaluate the stability of the material.Methods The reference regarding the target sequence for CT PCR detection has been reviewed.The ovedap extension technique and molecular cloning techniques were used to construct a recombinant lasmid.Then the recombinant plasmid pTARGETTM-CT was transfected into a HTB-SiHa cells.The cuhured epithelia cells,vere collected as quality control material.Then we evaluated the stability of this material with domestic kits for CT PCR detection.The stability in different conditions were summarized and evaluated.The EQA samples for CT test survey ere prepared from the above prepared cells and distributed to the EQA participants nationwidely.Results Five fragments from CT(178-610),(1219-1993),(2471-3260),(5239-5864),(6722-7499) were cloned into pTARGET TM.The recombinant plasmid was transfected into mammalian cells as a final form for the quality control materials.Real-time PCR analysis howed the original material was positive with domestic chlamydia trachomatis kit(3.21×108 copies/ml).A Series of dilution resulted in the decreased result .The stability testing indicated the quality control materials were stable at least for one month when stored at 4℃,room temperate or 37℃.Conclusions We used several kinds of molecular iology methods such as ovedap PCR and enzyrnatie digest to construct a recombinant plasmid which contained several fragments.The quality control materials for chlamydia trachomatis PCR detection was developed successfully.

[Objective] To research the sedative effect of intranasal dexmedetomidine and sufentanil for pediatric sedation for stomatological operation of outpatient department.[Methods] 60 children undergoing stomatological operation of outpatient department,age 3 ～ 7 years,weighing 10 ～ 32 kg,of ASA physical status Ⅰ ～ Ⅱ,were divided into the three groups (n =20) randomly using a random number table:group dexmedetomidine and sufentanil (group DS),group dexmetomidine (group D),group sufentanil (group S).Recorded the children's behavior using the Ohio State University behavior rating score (OSUBRS),the University of Michigan Sedation Score (UMSS),SBP,HR and side-effects when entry,during and leave operation and in post-anesthesia care unit,side-effects,the satisfaction of stomatological doctors and parents.[Results] The OSUBRS of group DS when entry,during operation were lower than group D (P ＜ 0.01).The UMSS of group DS were higher than group D and group S when entry and during operation (P ＜ 0.05).The success rate of group DS was higher than group D and group S (P ＜ 0.01).There was no different of mean percentage change in systolic blood pressure and heart rate from baseline between group DS and group D (P ＞ 0.05).There were no instances of respiratory depression,hypotension and bradycadia.[Conclusion] Intranasal dexmedetomidine and sufentanil provides satisfactory pediatric sedation for stomatological operation of outpatient department without side effects such as respiratory depression,bradycadia and hypotension.

Objective To investigate the effect of repetitive transcranial magnetic stimulation(rTMS)on the spinal segmental excitability after spinal cord injury in adult rats.MethodsT 10 spinal cord injury models were made with weight-drop method.8 weeks later,rTMS were applied to the experimental group at 0.5 Hz suprathreshold stimulation,500 pulses daily for 4 weeks.Spinal cord injury rats without stimulation and normal rats were used as controls.At different time points,electronic evoked F-wave were measured.The ratio of F-wave amplitude to M-wave amplitude(F/M)were compared among these groups.Immunohistochemistry was used to detect the expression of 5-hydroxytryptamine(5-HT)in the rostral and caudal lesion segments.ResultsThe ratio of F/M increased significantly(P<0.01)8 weeks after spinal cord injury compared with baseline ratio and regressed significantly(P<0.01)after 4 weeks of rTMS.Expression of 5-HT in grey matter around lesion was decreased after spinal cord injury and increased significantly(P<0.01)both in the rostral and caudal lesion segments in rTMS treatment group.ConclusionThe increased spinal segmental excitability after spinal cord injury can be regressed by rTMS,which may be resulted in increased expression of 5-HT.

Objective: To observe the clinical efficacy of Zhi Shen Tiao Sui (regulating the mind and marrow) needling in treating depression after ischemic stroke. Methods: A total of 145 patients with depression after ischemic stroke were recruited between January 2016 and January 2018 and divided into two groups using random number table method, with 72 cases in the control group and 73 cases in the observation group. The control group received conventional treatment for ischemic stroke and medication, while the observation group was given additional Zhi Shen Tiao Sui acupuncture. The two groups were treated for 4 successive weeks. Clinical efficacy was analyzed after treatment. The neurological function pertinent scores [Hamilton depression scale (HAMD), clinical neurological deficit score (CNDS), activities of daily living (ADL), Fugl-Meyer assessment scale (FMA) and Barthel index (BI) scores], as well as the levels of cortisol (CORT) and adrenocorticotropic hormone (ACTH) were recorded in the two groups. Adverse reactions that occurred during treatment were also observed. Results: The clinical total effective rate was 82.2％ in the observation group, significantly higher than 66.7％ in the control group (P<0.05). After treatment, the HAMD and CNDS scores were markedly reduced in both groups (all P<0.05), and the ADL, FMA and BI scores were markedly increased (all P<0.05). The HAMD and CNDS scores were significantly lower in the observation group than in the control group after treatment (both P<0.05), and the ADL, FMA and BI scores were significantly higher in the observation group than in the control group (all P<0.05). The levels of CORT and ACTH declined significantly after treatment in the two groups (all P<0.05), and were markedly lower in the observation group than in the control group (both P<0.05). The adverse reaction rate was 2.7％ in the observation group, versus 4.2％ in the control group, and the between-group difference was statistically insignificant (P>0.05). Conclusion: Compared with sole use of Western medication, medication combined with Zhi Shen Tiao Sui acupuncture can promote the recovery of neurological function, down-regulate the levels of CORT and ACTH and enhance the clinical efficacy in treating patients with depression after ischemic stroke, with a relatively high safety rating.

Objective To explore the effect of repetitive transcranial magnetic stimulation(rTMS)on spinal cord injured rats.Methods Weight-drop spinal cord injury model was made at thoracic 10 segments with NYU impactor device.Stimulated group received daily superthreshold rTMS continued for 4 weeks.BBB locomotor scores were recorded weekly.Growth associated protein 43(GAP43)and 5-hydroxytryptamine(5-HT)were detected with immunofluorescence staining in the area of rostral and caudal to the lesion.Results The BBB scores in stimulation group improved compared with that in the control(P<0.01).GAP43 and 5-HT markers increased in the stimulation group(P<0.01),and they increased in the rostral than in the caudal areas(P<0.01).Conclusion rTMS can improve the locomotor function of incomplete spinal cord injury rats,which may result from the increase of expression of GAP43 and 5-HT.

Objective To invespigape phe relapionship bepween plasma ospeoponpin levels and phe seveript of coronart apherosclerosis and ips predicpive value in phe diagnosis and prognosis of coronart arpert disease(CAD) . Methods 788 individuals were included in phis reprospecpive spudt. Thet underwenp coronart angiographt bepween Jan. 1, 2011 po Dec. 31, 2011. Thet were divided inpo five groups based on phe resulps of coronart angiographt: normal coronart, coronart apherosclerosis, 1-vessel disease, 2-vessel disease, 3-vessel ± lefp main disease. The plasma ospeoponpin concenprapions were measured bt ELSIA. The plasma ospeoponpin levels bepween differenp groups were compared. The areas under phe ROC curve (AUC) for plasma ospeoponpin levels were generaped po analtze phe predicpive value in phe diagnosis of coronart arpert disease. The clinical condipions were followed-up. Results There were 788 individuals included in phe spudt. The mean plasma ospeoponpin concenprapions of phese five groups were (37. 05 ±15. 23)μg/ L for normal coronart, (51. 01 ± 18. 81) μg/ L for coronart apherosclerosis, (66. 26 ± 23. 22) μg/ L for 1-vessel disease, (76. 92 ± 26. 39) μg/ L for 2-vessel disease and (88. 14 ± 28. 93) μg/ L for 3-vessel ± lefp main disease respecpivelt. The correlapion coefficienps of phe plasma ospeoponpin levels po phe number of damaged coronart vessels was 0. 511. The AUC for plasma ospeoponpin levels predicping CAD was 0. 821. The AUC for phe six pradipional risk facpors of coronart apherosclerosis predicping CAD was 0. 692. During phe follow-up, 79 subjecps (20. 1% ) wiph plasma ospeoponpin levels no higher phan 71. 55 μg/ L experienced endpoinp evenps, and 118 subjecps (29. 9% ) wiph plasma ospeoponpin levels higher phan 71. 55 μg/ L experienced endpoinp evenps (P =0. 001). Conclusions Plasma ospeoponpin levels were elevaped progressivelt wiph phe seveript of coronart arpert lesions. Plasma ospeoponpin levels had good predicpive value in phe diagnosis of coronart arpert disease and matbe a predicpor for cardiovascular evenps.

Aim The mechanism of basic fibroblast growth factor(bFGF)in mediating increase of intracellular free magnesium ([Mg2+]i) in human umbilical vein endothelial cells (HUVECs), and the relationship between Mg2+and angiogenesis were investigated in this study.Methods The change of[Mg2+]i in HUVECs were quantitatively detected in intracellular cation measurement system via loaded with the fluorescent magnesium indicator mag-fura-2. Endothelial cells were primarily acquired by infusion of collagen enzyme solutioninto the lumens of human umbilical veins and cultured in M199 with 0.2 fetal bovine serum. The role of bFGF in angiogenesis was observed in presence of 0,1 mmol?L-1 or 2 mmol?L-1 of extracellular Mg2+.Results bFGF dose-dependently increased [Mg2+]i, and there was not any significant difference among the groups of 0,1 mmol?L-1and 2 mmol?L-1 of extracellular Mg2+;similar results were obtained in groups done with Na+ and Ca2+. Pretreatment with bFGF receptor-2 (KDR) inhibitor (SU1498) blocked the increase of [Mg2+]i induced by bFGF.Unlike in the group of 0 mmol?L-1extracellular Mg2+,the apparent angiogeneses were observed in the groups of 1 mmol?L-1 and 2 mmol?L-1 extracellular Mg2+ in the presence of bFGF.bFGF-induced angiogenesis was significantly blocked with SU1498 in the presence of 1 mmol?L-1 extracelluar Mg2+.Conclusions These results suggest that the increase of [Mg2+]i by bFGF come from intracellular Mg2+ pools mediated by KDR-dependent signaling pathways,thereby resulting in the bFGF-induced angiogenesis.

Objective To evaluate the accuracy of air column width difference (ACWD) between inflation and deflation of the cuff of the endotracheal tube (ETT) in predicting post-extubation stridor (PES).Methods A total of 102 intubated patients of both sexes and all ages,who were mechanically ventilated for ≥24 h in the intensive care unit,of American Society of Anesthesiologists physical status Ⅲ or Ⅳ,were enrolled in the study.After the patients were in a stable condition and recovered consciousness and myodynamia,they were weaned from the ventilator,and ultrasound examination of the larynx was performed to determine ACWD between ETT cuff inflation and deflation.PES was assessed using blinding nethod after extubation,and the patients were divided into 2 groups depending on whether or not the patients developed PES:PES group (group P) and non-PES group (group N).Results There were 94 patients in group N and 8 patients in group P.Compared with group N,ACWD was significantly decreased in group P (P＜ 0.05).The cut-off value of ACWD determined by the receiver operating characteristic curve was 1.65 mm,the sensitivity and specificity of ACWD were 0.830 and 0.750,respectively,and the area under the receiver operating characteristic curve was 0.801.Conclusion For the intubated patients who are mechanically ventilated for ≥24 h in an intensive care unit,ACWD between ETT cuff inflation and deflation＜ 1.65 mm can effectively predict PES.

Objective To evaluate the expression of RAR-? gene in cervical carcinoma cell lines SiHa,HeLa,C33A and Caski and to analyze the relation between their gene expression and the promoter methylation of RAR-? DNA.Methods The expression of mRNA and protein of RAR-? gene in the four cell lines were analyzed by RT-PCR,western blot and immunofluoreseence,respectively.Methylation specific PCR(MSP)was used to check whether there was methylation in the promoter of RAR-? gene.The demethylating agent 5-aza-2'-deoxycytidine(5-Aza-cdR)was used to treat methylated cell lines and the change of RAR-? gene methylation and RAR-? gene expression defects were observed.The cell proliferation was assayed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.Results The mRNA and protein expression levels of RAR-? in cell lines SiHa,HeLa,Caski and C33A were 0.25 ?0.08,0,0.60?0.19,3.12?0.92 and 0.23?0.07,0,0.14?0.05,0.68?0.21,respectively.The mRNA and protein expression of RAR-? in SiHa,HeLa and Caski cell lines were decreased or silenced, whereas its expression increased in C33A cell line.MSP method showed that there were RAR-? gene methylation in SiHa,HeLa and Caski cell lines,while there was no RAR-? gene methylation in C33A cell line.After treated with 5-Aza-cdR,the mRNA and protein expression levels of RAR-? in SiHa,HeLa, Caski and C33A cell lines were 1.82?0.59,2.13?0.62,1.67?0.43,2.95?0.89 and 0.69?0.21, 0.83?0.29,0.56?0.16,0.64?0.20 respectively.The mRNA and protein levels of RAR-? had a significant difference between before and after interference with 5-Aza-cdR in SiHa,Helm,and Caski cell lines(P0.05).The 5-Aza-cdR treatment could suppress cell proliferation.Conclusions The RAR-? gene expression defects play an important role in the carcinogenesis of cervical cancer.Aberrant methylation in promotor region of RAR-? gene may be an important mechanism for the loss of expression of RAR-? gene.

Objective To evaluate the changes of the upper airway of the patients with obstructive sleep apnea syndrome (OSAS) before and after operations and to know the effects of operations by MSCT. Methods The upper airway dimensions of 26 patients with OSAS were measured on multiplanar reformatted (MPR), curved-planar reformatted (CPR), volume rendering(VR) images of 16-slice spiral CT. The measurements include the anteroposterior calibres and the areas on the reformatted axial images on the pharyngeal cavity levels, the calibres and the minimum areas in retropalatal and retroglossal regions, the areas of the soft palate and uvula on the reformatted sagittal view with maximum thickness, the maximum wall thickness of the right and left the upper airway on the coronary images, the volume of the upper airway before and after the operations. The measurements were correlated with the polysomnography (PSG) records. The data were analyzed paired-samples t-test and Pearson correlations. Results By comparison, the anteroposterior calibres and the cross-sectional areas on the reformatted axial view of the lower retropalatal region (slice 4) of the upper airway increased significantly after operations. The anteroposterior diameter increased from 5. 9 mm before operations to 12.8 mm after operations, where t = - 5.506, P < 0.05. The areas increased from 51.0 mm~2 before operations to 275.0 mm~2 after operations, where t = -5.011, P <0.05. In the higher retropalatal region (shce 2) of the upper airway, the anteroposterior diameter decresased from 14.8 mm before operations to 9.2 mm after operations, where t = 2.867, P < 0.05. The areas decreased from 241.0 mm~2 before operations to 128.0 mm~2 after operations, where t = 3.087, P < 0.05. The anteroposterior calibres of retroglossal region (slice 7) decreased from 12.7 mm before operations to 10.3 mm after operations,where t = 3.718, P <0.05. The L-R calibres and the minimum areas of of retropalatal increased significantly from 6.4 mm, 33.0 mm~2 before operation to 10.9 mm, 76. 0 mm~2 after operation, where t = -3.413, -2. 216, respectively and P < 0.05. Of the 9 cases whose apnea and hypopnea index (AHI) ≤5 events/hour after operations, the minimum areas of retropalatal region, the anterio-posterior diameter, L-R calibres increased significantly. The areas increased from 41.0 mm2 before operations to 76.0 mm~2 after operations, were t = -4. 932, P <0.05. The anteroposterior calibres increased from 4.6 mm before operations to 6.6 mm after operations, where t = - 7. 308, P < 0.05. The L-R calibres increased from 8.3 mm before operations to 13.6 mm after operations, where t = - 4.320, P < 0.05. Conclusions MPR、CPR、VR of MSCT can evaluate the not only the morphology but the function changes of the upper airways on the OSAS patients. The increasing of the minimum cross-sectional area may be one of the important indications for evaluating operations. The narrowing of the higher retropalatal region of the upper airway after operations should be an alert to the clinicians.