Objective To clone,express,and purify Tp 0319 outer membrane protein of Treponema pallidum and to develop an indirect ELISA for diagnosing syphilis.Methods The expression plasmid PQE32/Tp 0319 was conventionally constructed.The recombinant Tp 0319 protein was produced in E.coli M15 after induction by IPTG.The Tp 0319 protein was analyzed by SDS-PAGE and Western blotting,and then purified with Ni-NTA affinity chromatography.Indirect ELISA was developed to detect the syphilis antibody in human sera.Results The recombinant plasmid PQE32/Tp 0319 was constructed successfully and the fusion protein with relative molecular weight near 30 000 Dalton was revealed by SDS-PAGE.Western blotting proved that the recombinant protein specifically reacted with anti-Tp antibodies in sera from syphilis patients.The results of the indirect ELISA indicated the sensitivity and the specificity were both 100%.The concordance of 300 sera(150 from blood donors and 150 from syphilis patients)detected in parallel by the ELISA and the TPPA was 95.3%.Conclusions The data suggest that the prepared recombinant protein Tp 0319 of Treponema pallidum has high immunoreactivity.The recombinant protein can be used to develop ELISA kit for diagnosing syphilis.

ObjectiveTo observe the effects of hypoxia inducible factor-1 alpha (HIF-1α)-targeting small interfering RNA(siRNA) on the expression of HIF-1α and vascular endothelial growth factor (VEGF) in HaCaT ceils under hypoxic conditions. MethodsHaCaT cells were cultured and divided into four groups, normal control group (without any treatment), hypoxia group (cultured under hypoxic conditions for 24 hours),liposome control group (transfected with liposome followed by hypoxic culture for 24 hours), RNA interference group (transfected with HIF-1α-targeting siRNA/liposome complexes followed by hypoxic culture for 24 hours). Fluorescence real-time quantitative PCR was utilized to determine HIF-1oα and VEGF mRNA expression in HaCaT cells, and Western blot to detect HIF-1α and VEGF protein expression. ResultsNo significant difference was observed in the mRNA expression of HIF-1α between the hypoxia group and normal control group(0.907 ± 0.032 vs. 0.878 ± 0.034, F =1.108, P ＞ 0.05), while the expression levels of VEGF mRNA,HIF-1α and VEGF proteins were significantly higher in the hypoxia group than in the normal control group (0.935 ± 0.032 vs. 0.652 ± 0.053, 0.813 ± 0.047 vs. 0.236 ± 0.014, 0.791 ± 0.030 vs. 0.316 ± 0.013, all P ＜0.05). A significant decline was noted in the mRNA and protein expression levels of VEGF (0.230 ± 0.044 vs.0.978 ± 0.030, 0.213 ± 0.026 vs. 0.817 ± 0.049, both P ＜ 0.05) and HIF-1α(0.497 ± 0.033 vs. 0.806 ±0.040, 0.249 ± 0.028 vs. 0.833 ± 0.052, both P ＜ 0.05) in the RNA interference group than in the liposome control group. ConclusionsHypoxia may enhance the expression of HIF-1α and VEGF in HaCaT cells, and to inhibit the HIF-1α expression may suppress the expression of VEGF in HaCaT cells under hypoxia.

Objective To investigate the prevalence of A2058G or A2059G mutation within 23S rRNA in Treponema pallidum (Tp) from primary syphilis patients chancre samples. Methods Simple PCR was used to screen the positive samples containing Tp DNA. Nested PCR was adopted to amplify the region of the Tp 23S rRNA and the purified amplicons were digested by restriction endonuclease MboⅡand Bsa I respectively and sequenced. Results 39 qualified samples were obtained from 43 chancre samples and all of them were found harboring the A2058G mutation, whereas the A2059G was not detected. Conclusion High frequency of the A2058G mutation within 23S rRNA implicated in macrolide resistance emerges in the circulating Tp in Hengyang. Therefore, macro-lide antibiotics such as azithromycin should be cautiously used as an optional therapy for syphilis.

Objective To construct a recombinant plasmid encoding Tp0821,a membrane lipoprotein of T. pallidum,express and purify this protein,and to evaluate its immunocompetence.Methods The recombinant plasmid pQE32/Tp0821 was constructed and induced to express the corresponding protein.Then,New Zealand rabbits were immunized with purified recombinant protein to prepare polycional antibodies,and the titer of polyclonal antibody was determinated.Indirect ELISA was developed with the recombinant protein of T. pallidum as coating antigen to detect 80 control sera and 150 FTA-ABS-positive sera.Results The recombinant plasmid pQE32/Tp0821 was constructed and a fusion protein with expected molecular weight was expressed.Specific humoral response was elicited by the recombinant protein in New Zealand rabbits and the antibody titer reached 1:6400.Compared with FTA-ABS test,the indirect ELISA showed a sensitivity and specificity of 92.6%and 98.6%,respectively,in the detection of control and clinical sera.Conclusion The recombinant protein Tp0821 shows excellent immunocompetence,which can be applied to the serological diagnosis of syphilis.

Objective To clone, express Tp0319 gene from Treponemapallidum (T. pallidum), and to assess the immunocompetence of recombinant protein. Methods The immuno-dominant region of Tp0319gene was chosen by computer analysis, amplified from T. pallidum complete genome by PCR, subcloned into the expression vector pQE32 to construct a recombinant plasmid, pQE32/Tp0319, which was then expressed in E. coli M15. The recombinant protein was purified with Ni-NTA affinity chromatography, and identified by using sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western blot. New Zealand rabbits were immunized with the recombinant protein, and the titer of anti-Tp0319 antibodies in sera from immunized rabbits were measured with indirect ELISA. Also, indirect ELISA with the recombinant Tp0319 as coating antigen was performed to detect the anti-Tp0319 antibody in sera from 200 normal human controls and 200 patients with syphilis. Results The prokaryotic expression vector pQE32/Tp0319 was constructed successfully, and the recombinant protein Tp0319 with a molecular weight of about 30 000 was attained. Specific humoral response was elicited by the recombinant protein in New Zealand rabbits and the specific antibody titer was more than 1: 10 240 after immunization for 3 times. Western blot proved that the recombinant protein could specifically react with anti-T. pallidum IgG antibody-positive sera. Indirect ELISA was successfully developed with the recombinant Tp0319, and detected antibodies to T. pallidum in control sera with a sensitivity and specificity of 100% (40/40), respectively. Compared with T. pallidum particle agglutination (TPPA) assay, the sensitivity and specificity of the indirect ELISA were 92.6% and 100%, respectively, in the detection of T. pallidum in sera from patients and controls, and the concordance between the indirect ELISA and TPPA was 96%. Conclusions The prepared recombinant protein shows a satisfactory immunocompetence, which may lay a foundation for its further application in the serodiagnosis of syphilis.

Objective To study the clinical manifestation of intracranial anomalous venous return associated with the facial nevi and hydrocephalus. Methods Along with reviewing of the literatures, the clinical records of 3 patients suffered from the facial nevi and hydrocephalus were analyzed. Results All of 3 patients also have sinovenous occlusion, which affect cerebral venous return. Conclusions Intracranial anomalous venous return, which is induced by extensive sinovenous occlusion, has been considered to be the main cause of hydrocephalus in the facial nevi. The formation of sinovenous occlusion and anomalous venous return may be due to developmental defect in the embryonic stage.

The paper analyzed the challenges for community-based rehabilitation of schizophrenic patients in China' s rural areas,from such six aspects as culture, prevention and control system for mental health, fairness of health resources allocation, shortage of specialists, confidence in treatment,and delay in rehabilitation. These studies aim at providing the government with decision making evidence for enacting rural mental health policies and taking effective intervention measures.

Objective: To explore the application value and safety of low-dose dobutamine stress echocardiogram (LDDSE) in patients of low-flow/low-gradient aortic stenosis combining left ventricular dysfunction with transcatheter aortic valve replacement (TAVR). Methods: A total of 5 eligible consecutive patients with contradiction of routine surgical valve replacement and going to receive TAVR in our hospital from 2013-10 to 2016-07 were enrolled. The mean aortic valvegradient, maximum flow velocity, each stroke volume and ejection fraction were recorded before and during LDDSE examination. The patients having confirmed diagnosis of true severe aortic stenosis with left ventricular contractile reserve received TAVR, for those without left ventricular contractile reserve received drug therapy or TAVR conditionally. The changes of cardiac function and NT-proBNP level were observed after TAVR. Results: All 5 patients showed positive finding in LDDSE; the mean aortic valve gradient ≥40mmHg and stroke volume≥20% implied that the patients had true severe aortic stenosis with left ventricular contractile reserve. No adverse reaction occurred during and after LDDSE. TAVR was performed in 4 patients and 1 was waiting for TAVR or balloon dilatation since temporary lacking of valve. The post-operative cardiac function was improved in all patients and NT-proBNP level was declined continuously. Conclusion: LDDSE examination could be considered in patients of aortic stenosis combining left ventricular dysfunction, low-flow and low-gradient to clarify ventricular contractile reserve and the severity of aortic stenosis. If the patients with ventricular contractile reserve, TAVR was recommended which was the effective treatment for relevant patients.

Objective:To investigate the clinical significance of soluble growth stimulating expression gene 2 protein (sST2) combined with neutrophil/lymphocyte ratio (NLR) in the prediction of nosocomial cardiovascular adverse events in patients with acute carbon monoxide poisoning (ACOP) myocardial injury.Methods:Patients with ACOP myocardial injury from January 2017 to December 2019 in Emergency Ward and EICU of Harrson International Peace Hospital, Hebei Medical University were enrolled. NLR was calculated by routine blood examination on admission, and sST2 (T 0sST2, T 3dsST2) was detected by ELISA on admission and at 3 days after admission. According to the occurrence of cardiovascular adverse events, the patients were divided into the event group and the non-event group. Logistic regression was used to analyze the risk factors of in-hospital cardiovascular adverse events. ROC curve was used to analyze the value of sST2, NLR, sST2 and NLR combined in predicting the occurrence of in-hospital cardiovascular adverse events in patients with ACOP myocardial injury. Results:Totally 255 patients with ACOP myocardial injury were included in the final analysis. NLR was (13.38±4.33) in the event group and (9.57±4.22) in the non-event group, T 3dsST2 was (61.59±22.67) ng/mL in the event group and (40.52±13.14) ng/mL in the non-event group, with statistically significant differences (all P<0.01). T 0sST2 was (265.34±89.95) ng/mL in the event group and (242.43±93.09) ng/mL in the non-event group, with no statistically significant difference ( P=0.333). Logistic regression analysis showed that NLR ( OR=1.270, 95% CI: 1.125-1.434, P<0.01) and T 3dsST2 ( OR=1.082, 95% CI: 1.052-1.114, P<0.01) were independent risk factors for nosocomial cardiovascular adverse events in patients with ACOP myocardial injury. The optimal cutoff value of T 3dsST2 was 44.5 ng/mL, and of NLR was 12.08. The sensitivity and specificity of dual T 3dsST2 and NLR in predicting nosocomial cardiovascular adverse events was 79.3% and 82.7%, respectively (AUC 0.857, Youden index 0.620). Conclusions:T 3dsST2 and NLR are independent risk factors for the nosocomial cardiovascular adverse events in patients with ACOP myocardial injury. The predictive cutoff values are 44.5 ng/mL for T 3dsST2 and 12.08 for NLR. Combination of T 3dsST2 and NLR has a practical predictive value for nosocomial cardiovascular adverse events in patients with ACOP myocardial injury.

ObjectiveTo observe the effect of rhubarb as the main composition in the therapy for patients with acute paraquat poisoning (APP).MethodsA prospective randomized controlled trial was conducted. 128 patients with APP admitted to Harrison International Peace Hospital from March 2011 to December 2013 were randomly divided into western medicine control group (n = 64) and western medicine and traditional Chinese medicine (TCM) combination group (n = 64). All the patients were treated with repeated gastric lavage and oral kaolin. The patients in western medicine control group were given 20% mannitol and (or) magnesium sulfate for catharsis, early (within 8 hours of admission) hemoperfusion (HP), and also given the routine combined therapy. In TCM combination group, in addition to the above treatment patients were given oral paraquat poisoning detoxification prescription No.1 every 2 hours for catharsis, which was composed of rhubarb 10 g, glauber salt 12 g, agrimony 12 g, and licorice 6 g. When green stool disappeared, detoxification therapy was changed to No. 2 compound once a day for 14 days, which was consisted of rhubarb 10 g, ginseng 6 g, agrimony 15 g, rhizoma chuanxiong 10 g, licorice 6 g. The poison volume, first dose of oral drug, time for the first HP, time of the first defecation, the time of last green stool, decontamination time, white blood cell count (WBC), C-reactive protein (CRP), arterial blood gas analysis, blood lactic acid (Lac), liver and kidney functions, myocardial enzyme spectrum, chest CT, adverse reaction, days of hospitalization, and mortality rate were observed in both groups. The levels of paraquat in plasma and urine were determined before treatment and 12 hours after poisoning in both groups. Sixty days after discharge, chest CT was taken for observation of pulmonary fibrosis. ResultsThere were no significant differences in the poison volume, ingestion time and the time for the first HP between the two groups. Compared with western medicine control group, the first defecation time (hours: 3.94±1.14 vs. 6.17±1.52), the last time of green stool (hours: 36.90±4.10 vs. 51.63±4.91), and poison clean-up time from plasma (hours: 19.48±3.63 vs. 23.84±3.29) in combination with TCM group were significantly earlier (allP< 0.01). WBC, CRP, alanine aminotransferase (ALT), blood urea nitrogen (BUN), and serum creatinine (SCr) levels were gradually increased after admission in combination with TCM group, and they peaked on 5th day [WBC (×109/L): 15.35±2.17 vs. 17.47±2.09, CRP (mg/L): 32.62±2.76 vs. 39.51±2.45, ALT (U/L): 270.88±11.06 vs. 334.67±7.85, BUN (mmol/L): 13.29±1.90 vs. 17.63±1.42, SCr (μmol/L): 203.54±18.40 vs. 251.53±14.38, allP< 0.05], and then they declined; Lac after admission was gradually increased, and peaked at 7 days (mmol/L: 3.53±0.21 vs. 3.82±0.14, P< 0.05), and then it was decreased. Myocardial enzyme spectrum was increased after admission, and peaked on 3rd day [creatine kinase (CK, U/L): 192.09±16.26 vs. 216.20±11.96, creatine kinase isoenzyme enzyme (CK-MB, U/L):39.03±3.75 vs. 47.22±5.84, bothP< 0.05), and then they declined gradually. Arterial partial pressure of oxygen (PaO2) and base excess (BE) were gradually decreased after admission, down to trough on the 7th day after admission [PaO2 (mmHg, 1 mmHg = 0.133 kPa): 87.04±2.37 vs. 84.93±2.44, BE (mmol/L): -7.31±2.31 vs. -9.18±2.49, bothP< 0.05], and then they were increased. At 12 hours after poisoning, paraquat contents in plasma and urine in combination with TCM group were significantly lower than those of the western medicine control group [plasma (ng/L):0.83±0.08 vs. 0.96±0.10, urine (ng/L): 0.88±0.09 vs. 0.97±011, bothP< 0.05]. The injury to lung tissue was significantly improved in combination with TCM group compared with that in the western medicine control group, and no serious adverse reactions was found, and the hospital stay time (days: 20.46±6.07 vs. 29.73±9.16) was significantly shortened (P< 0.01), and the mortality rate [35.9% (23/64) vs. 45.3% (29/64)] was significantly lowered compared with western medicine control group (P< 0.05). In the combination with TCM group pulmonary fibrosis was lighter than that in the western medicine control group during the 60-day follow-up. ConclusionThe sequential treatment of No. 1 and No. 2 detoxification of APP, using rhubarb as the main drug, can effectively eliminate paraquat, reduce absorption of the toxin, prevent the development of multiple organ dysfunction syndrome (MODS) induced by the toxin, shorten the hospital stay days, and improve the prognosis of APP.