Objective To observe the impact factors of the diagnostic accuracy of fine needle aspiration biopsy(FNAB)for papillary thyroid carcinoma(PTC).Methods Totally 468 patients with single PTC confirmed by postoperative pathology who underwent FNAB before surgery were enrolled.The impact of clinica,l ultrasonic and pathological features on the accuracy of FNAB diagnosis were analyzed.Results The accuracy of FNAB for diagnosing PTC was 71.37%(334/468).The maximum diameter and location of PTC were both impact factors of the diagnostic accuracy of FNAB.The maximum diameter of 0.7 cm was the optimal cutoff value of FNAB for diagnosing PTC,and the diagnostic accuracy of FNAB for PTC with the maximum diameter＜0.7 cm and those≥0.7 cm was 62.96%(119/189)and 77.06%(215/279),respectively.The diagnostic accuracy of FNAB for PTC located in the difficult and easy area of puncture was 52.53%(52/99)and 76.42%(282/369),respectively.The diagnostic accuracy of FNAB for PTC with the maximum diameter≥0.7 cm and located in the easy area,≥0.7 cm and located in the difficult area,＜0.7 cm and located in the easy area,＜0.7 cm and located in the difficult area was 80.43%(185/230),61.22%(30/49),69.78%(97/139)and 44.00%(22/50),respectively.Conclusion The maximum diameter and location of PTC were both impact factors of the diagnostic accuracy of FNAB.

Objective: To investigate the concentration of serum bone resorption markers NTX and TRACP-5b of children aged 8-14 years in a coal-burning fluorosis area and its relationship with age, and to provide population data for the study of pathogenesis of skeletal fluorosis. Methods: Totally 123 children of 8-14 schoolage in the two primary schools in Doujing Township, Shuicheng County, Liupanshui City, Guizhou Province were randomly selected as the exposed group. According to the matching principle, 64 children were randomly selected as a control from a primary school in a nondisease area Huaga Town. The dental fluorosis was investigated, and the concentrations of serum NTX and TRACP-5b were measured. Results: The detection rate of dental fluorosis in the fluorosis area was 94.3% and 0 in the control area. The concentrations of serum NTX in fluorosis area children were 13.04 (10.76, 15.64), 14.82 (12.15, 18.26)nmol/L in the early adolescence and middle-aged period, which lower than the control area 15.73(14.36, 18.61), 16.45(15.45, 22.02)nmol/L( P <0.05); The serum TRACP-5b levels in children with fluorosis were 276.74(237.63, 312.75), 270.14(242.82, 321.97), 305.95(259.78, 339.87)nmol/L in prepubertal, early adolescence and middle youth, lower than the control area 370.88 (304.47, 452.84), 353.30 (262.05, 393.19), 420.22 (376.96, 544.60)nmol/L( Z =-3.03, -2.66, -3.10, P <0.05). Serum NTx and TRACP-5b in fluorosis area were negatively correlated with dental fluorosis in children( r =-0.51, -0.37, P <0.01). Conclusion Fluorosis can reduce the concentrations of serum bone resorption markers NTX and TRACP-5b in children of different age groups. TRACP-5b may be more sensitive to fluoride exposure than NTX, but the specific mechanism remains to be further studied.

Objective To explore the diagnostic value of thyroglobulin measurement with fine‐needle aspiration ( FNA‐Tg ) in detecting papillary thyroid carcinoma ( PTC ) metastatic lymph nodes ,and analyze the impact of different factors on the diagnostic accuracy of FNA‐Tg . Methods One hundred and fifty‐four lymph nodes from 138 patients w ho underwent total thyroidectomy and cervical lymphadenectomy were enrolled . Ultrasonography ,fine needle aspiration cytology ( FNAC) and FNA‐Tg results were acquired in all patients . T he diagnostic value of FNAC ,FNA‐Tg and the combination of the two methods were compared in suspicious PTC metastatic lymph nodes . T he influence of lymph nodes size ,lymph nodes cystic change and serum thyroglobulin antibody( TgAb) on the accuracy of FNAC and FNA‐Tg were analyzed . Results T he sensitivities of FNAC ,FNA‐Tg ,and the combination of the two methods in the diagnosis were 79 .5% , 95 .5% and 97 .7% ,respectively . T he specificities were 100% ,93 .9% and 100% ,respectively . T he size and cystic change of lymph nodes had effects on the accuracy of FNAC( P ＜0 .05) ,w hile they had no effect on the accuracy of FNA‐Tg ( P ＝0 .42 ,0 .67 ) . T he serum TgAb level had no influence on the accuracy of FNAC and FNA‐Tg ( P ＝ 0 .88 ,0 .34 ) . Conclusions FNA‐Tg is sensitive to metastatic lymph nodes in PTC w hich not affected by the lymph nodes size ,the lymph nodes cystic change and serum TgAb level . Application of FNA‐Tg with FNAC can improve the sensitivity of diagnosis ,w hich is important in the diagnosis of PTC metastatic lymph nodes .

Objective To evaluate the efficacy of general anesthesia combined with ultrasoundguided epidural block for laparoscopic surgery in neonates.Methods Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ neonates,aged 20-60 days,scheduled for elective radical operation for Hirschsprung's disease,were divided into 2 groups (n=30 each) using a random number table:general anesthesia group (group Ⅰ) and general anesthesia combined with ultrasound-guided epidural block group (group Ⅱ).Anesthesia was induced by inhaling 8％ sevoflurane.Anesthesia was maintained with Ⅳ sufentanil 0.2 μg/kg,cisatracurium 0.1 mg/kg and then with additional sufentanil 0.1 μg/kg and cisatracurium 0.05 mg/kg every hour and 2％-3％ sevoflurane was simultaneously inhaled in group Ⅰ.In group Ⅱ,anesthesia was maintained by inhaling 2％-3％ sevoflurane,epidural block was performed at L1.2 interspace,the catheter was caudally advanced into the epidural space under ultrasound guidance,0.8％ lidocaine was injected at a loading dose of 6 mg/kg,the local anesthetic diffusion in the epidural space was observed,and additional 0.8％ lidocaine 3 mg/kg was given every 30 min.Heart rate and mean arterial pressure were recorded at 5 min before pneumoperitoneum,during pneumoperitoneum,at extubation and after extubation.The extubation time,duration of recovery room stay and development of assisted ventilation after extubation were recorded.Pain was assessed and scored at 5 min before discharge from recovery room.The development of epidural block-related complications was recorded in group Ⅱ.Results Operation was smoothly completed with stable anesthesia in two groups.Compared with group Ⅰ,the mean arterial pressure was significantly decreased during pneumoperitoneum and after extubation,the extubation time and duration of recovery room stay were shortened,and the requirement for assisted ventilation after extubation and pain scores were decreased in group Ⅱ (P＜0.05 or 0.01).No epidural block-related complications were found in group Ⅱ.Conclusion General anesthesia combined with ultrasound-guided epidural block is safe and effective for laparoscopic surgery and is helpful for postoperative recovery in neonates.

We constructed the CAP2NC prokaryotic expression vector of HIV-1 NL4-3 strain and obtained relatively pure CAP2NC protein by optimizing its purification conditions to explore its in vitro self-assembly conditions. Primers were designed according to the CAP2NC DNA sequence of HIV-1 NL4-3 strain. The target gene was amplified by PCR and cloned into prokaryotic expression vector pTO-T7. Then the recombinant strain was transformed into Escherichia coli BL21 (DE3). IPTG induced protein expression, then the protein was purified by hydrophobic chromatography. SDS-PAGE and Western blotting were performed to analyze the target protein, and the biological activity of the antigen was identified through ELISA. The self-assembly of CAP2NC protein was analyzed by transmission electron microscopy and gel filtration chromatography. The protein had good reaction with the specific antibodies of p24 and formed different structures in various conditions. When 10% yeast RNA was added to the protein complex, the recombinant protein only formed into a tubular structure, which was similar to the self-assembled structure of the HIV-1 virus capsid. The results showed that the HIV-1 CAP2NC protein had in vitro self-assembly activity, and the RNA affected the structure of CAP2NC protein assembly. The protein can be used as a simple and effective molecular model to study its structure, and then it can provide a reference for the study of HIV immature virus particles.

Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.

Objective To analyze the characteristics of clinical manifestations, risk factors, therapies and acute outcomes in patients with cerebral venous sinus thrombosis complicated by cerebral hemorrhage. Methods Seventy-five patients with cerebral venous sinus thrombosis were included in the study. According to the radiological findings on the brain image, patients were divided into two subgroups:cerebral hemorrhage group and non-hemorrhage group. The demo?graphic data, potential risk factors, clinical manifestations, radiological features, therapeutic strategies and acute out?comes were compared between two subgroups, and high risk factors were also analyzed. Results There were seventy-five patients with cerebral venous sinus thrombosis in the present study. Twenty-eight patients of them (37.2%) had cerebral hemorrhage whereas the remaining forty-seven patients (62.7%) did not have cerebral hemorrhage. Pregnancy/puerperi?um were significantly higher in patients with cerebral hemorrhage (with vs without;28.6%vs. 6.4%, P=0.015), while in?fection was markedly higher in patients without cerebral hemorrhage (with vs without;7.1% vs. 29.8%, P=0.021). Head?ache (92.9% vs. 70.2%, P=0.021), unconsciousness (25.0% vs. 6.4%,P=0.034), seizures (53.6% vs. 19.1%, P=0.002) and motor deficits (35.7% vs. 12.8%, P=0.019) were more common in patients with cerebral hemorrhage. Moreover, mul?tiple sinus involvement (1.4% vs. 44.7%, P=0.024) was significantly higher and the acute outcomes(mRS≥3: 46.4%vs.17.0%, P=0.006)were poorer in patients with cerebral hemorrhage. Binary Logistic analysis showed that pregnancy/pu?erperium (P=0.004) and multiple sinus involvement were positively, whereas infection was negatively correlated with cere?bral venous sinus thrombosis and hemorrhage ( P=0.007;P=0.03). Conclusions Pregnancy/puerperium, headache, uncon?sciousness, seizures, motor deficits and multiple sinus involvement are more frequently in patients with cerebral venous sinus thrombosis and hemorrhage, and the acute outcomes are poorer in patients with cerebral venous sinus thrombosis complicated by cerebral hemorrhage.

Objective To investigate the risk factors, diagnostic evaluation, etiology, and treatment in young patients with ischemic stroke. Methods The clinical data of young patients (age range 18 - 45 years) with ischemic stroke (within 2 weeks of stroke onset) admitted to the Stroke Center, the First Affiliated Hospital of Sun Yat-sen University from August 2008 to July 2013 were registered prospectively. Results A total of 300 patients were enroled, their age was 37. 8 ± 6. 8 years. There were 227 males (75. 7% ). The age of 84 patients (28. 0% ) was ≤35 years. The most common risk factors were smoking (43. 3% ), hypertension (38. 7% ), and hypercholesterolemia (38. 0% ). The positive result detection rates were lower in the antinuclear antibody (8/278), anticardiolipin antibody (34/250), 24-h Holter monitoring (2/60 ), and transthoracic echocardiography (38/232). According to the MRI findings, the detection rate of the white matter changes, old infarcts, single acute infarcts, and multiple acute cerebral infarcts were 40/282, 77/282, 145/282, and 137/282, respectively. The stroke subtypes of TOAST: large artery atherosclerosis (26. 7% ), smal artery occlusion (18. 0% ), cardioembolism (10. 0% ), other determined etiology (23. 0% ), and undetermined etiology (22. 3% ). Large artery atherosclerosis was mainly involved in anterior circulation (69/80) and intracranial arteries (75/80). In other definite causes, cerebral artery dissection was most common (36/69), and other causes included moyamoya disease (7/69) and infectious vasculitis (7/69), etc. The average length of hospital stay was 15. 4 d. The mean National Institutes of Health Stroke Scale score on admission was 7. 0, and the mean modified Rankin scale score at discharge was 2. 5. The complication rate during hospitalization was 9. 7% . 80. 3% and 48. 3% of patients received anti-platelet drugs and statins therapy during hospitalization.Conclusions This study used a prospective single-center method. It conducted a comprehensive analysis of risk factors, diagnostic evaluation, treatment, and etiology in current Chinese young patients with ischemic stroke. Its research data wil provide useful information for establishing a diagnostic strategy of high performance cost ratio, in-depth understanding of its pathophysiological mechanisms, and improving treatment strategies in Chinese young patients with ischemic stroke.

Objective Toinvestigatetheriskoflong-termrecurrenceofstrokeanditspredictorsin youngpatientswithischemicstroke/transientischemicattack(TIA).Methods Theclinicaldataofthe consecutive young patients (18-45 years)with ischemic stroke/TIA (within 2 weeks after onset)admitted to the department of neurology in the First Affiliated Hospital of Sun Yat-Sen University between August 2008 and July 2013 were enrolled prospectively. All patients were regularly followed up for a long time (The patients were followed up at the 1 st,6 th,and 12 th month after onset;then they were followed up once for every 6 months)in order to investigate stroke recurrence. The Kaplan-Meier curves were used to analyze the cumulative stroke recurrence rate of all patients. The last contact time for patients lost to follow was used as censored data to be enrolled in the analysis. The univariate analysis of the related risk factors for stroke recurrence using Log-rank test. Multivariate Cox proportional hazard regression was used to detect the related risk factors associated with stroke recurrence (adjusting for age and sex). The variables of the results of Log-ranktestP≤0.1wasselectedandenrolledinthemultivariateregressionanalysis.Results Atotalof 312 patients were enrolled in the analysis,including 294 with ischemic stroke and 18 with TIA. Their mean follow-up time was 34 ± 19 months. Thirty-four patients had recurrent stroke,including 23 with ischemic stroke,7 with TIA,and 4 with cerebral hemorrhage. The cumulative recurrence rates of stroke at 1 ,3 , and 6 years after onset were 6. 2%,10. 3%,and 16. 4%,respectively. The results of multivariate Cox proportional hazards regression analysis showed that hypertension (risk ratio [RR]2. 159;95% confidence interval [CI]1. 048-4. 447,P=0. 037)and cardioembolism (RR,2. 869;95%CI 1. 119-7. 357,P=0.028)weretheindependentpredictorsforstrokerecurrence.Conclusion Theoverall6-yearriskof recurrent stroke is not high in the Chinese young patients with ischemic stroke/TIA,but the risk of stroke recurrence is relatively higher in the first year. Hypertension and cardioembolism are the potential predictors of stroke recurrence;therefore,attention should be paid in clinical practice.

Objective To evaluate the value of a Treponema pallidum (TP) recombinant protein TP0993 in the serodiagnosis of syphilis.Methods A bioinformatics method was used to obtain the sequence of TP0993 gene.The open reading frame (ORF) without upstream non-coding region of TP0993 gene was ligated into the expression vector PET-28a (+),which was then transformed into Escherichia coli Rosetta.Isopropyl-β-d-thiogalactoside (IPTG) was used to induce the expression of TP0993 protein.The expressed protein was purified with nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography.Western blot was performed to evaluate the immunoantigenicity of the protein.New Zealand rabbits were immunized with the recombinant protein for immunogenicity evaluation.Indirect enzyme linked immunosorbent assay (ELISA) was developed by using the purified recombinant protein to coat microwell plates.Anti-TP antibodies were detected by the established ELISA and TP particle agglutination assay (TPPA) in 480 clinical serum samples.Results The prokaryotic expression vector PET-28a (+)-0993 was successfully built,and a fusion protein with a relative molecular weight of about 34 000 Da was attained after IPTG-induced expression and purification.Western blot proved that the recombinant protein could specifically react with clinical sera positive for anti-TP IgG antibodies.Specific humoral response was elicited in New Zealand rabbits by the recombinant protein.Compared with TPPA,the established indirect ELISA showed a sensitivity of 88.3％ and a specificity of 85.8％.There was a consistency of 86.5％ between the indirect ELISA and TPPA.Conclusion The expressed recombinant protein showed favorable immunocompetence,and may serve as a candidate antigen for serodiagnosis of syphilis.