Objective To investigate the feasibility and prospects of nested real-time PCR(NR-PCR)technique for Treponema palladium(Tp)detection in various samples of different stages of syphilis from patients preliminarily diagnosed as syphilis. Methods Targeting the Tp polA gene, NR-PCR was performed to detect Tp DNA in various samples from the patients with various stages of syphilis at the first clinic visit, including skin tissue fluid swabs, serum, whole blood, cerebrospinal fluid(CSF)and earlobe blood. Data were analyzed with SPSS software version 13. Results A total of 368 clinical samples were collected from 200 patients with syphilis. With a detection limit of 2 Tp/ml, NR-PCR showed that the total positive rate for Tp DNA was 71.7%(264/368). The Tp DNA positive rate was highest in earlobe blood samples (92.0%, 23/25), followed by CSF samples(90.2%, 46/51), skin tissue fluid swabs(74.3%, 26/35), serum samples(66.9%, 99/148)and whole blood samples(64.2%, 70/109). There was good agreement between NR-PCR results and serologic test results, with a consistency rate of 76.0%(152/200). Furthermore, the Tp DNA positive rate did not differ between patients with primary(12/19)and secondary syphilis(14/16)in skin tissue fluid swabs(χ2 = 2.62, P > 0.05), and was slightly but insignificantly higher in patients with secondary syphilis than those with primary syphilis in the serum samples(χ2=3.6, P=0.06). The Tp DNA positive rate of whole blood samples was also higher in patients with secondary syphilis than those with any other types of syphilis. Among patients with neurosyphilis, no significant difference was observed in the Tp DNA positive rate between earlobe blood samples and CSF samples(P=0.06). Among patients with latent syphilis, the Tp DNA positive rate was significantly higher in serum samples with an RPR titer of ≥ 1:8 than those with an RPR titer of≤1:4. Conclusion NR-PCR is feasible for detecting Tp DNA in various kinds of samples, and the Tp DNA positive rate is influenced by stages of syphilis and types of samples, as well as RPR titers.

Objective: To establish the quality standard for compound Heishen pills. Methods: Scrophulariae Radix, Radix et Rhizoma and Belamcancae Rhizoma were identified by TLC. HPLC was used to determine the content of harpagoside and cinnamic acid in Scrophulariae Radix on a Welchrom-C18 column using methanol-acetonitrile-1% ethylic acid (8∶21∶71) as the mobile phase. The flow rate was 1. 0 ml·min-1 . The column temperature was at 30℃ and the detection wavelength was set at 278 nm. Results:The TLC method had good specificity without interference from negative control. The linear range of harpagoside was 1. 32-65. 80μg·ml-1 with the average recovery of 98. 06%(RSD=2. 16%),and that of cinnamic acid was 0. 38-19. 20 μg·ml-1 with the average recovery of 98. 78%(RSD=1. 34%). RSDs of precision, stability and reproductibility tests were all below 2%. Conclusion: The established method is accurate, feasible and reproducible. It can be used in the quality control of compound Heishen pills.

Objective To evaluate the efficacy of T-joint endoscopy mask for fiberoptic bronchoscopy (FOB)-guided awake nasotracheal intubation in patients with cervical spinal cord injury.Methods Forty patients of both sexes aged 21-64 yr with fracture of cervical spine complicated by spinal cord injury scheduled for anterior decompression and interbody fusion under general anesthesia were randomly divided into 2 groups according to the technique for awake nasotracheal intubation (n =20 each):group nasal catheter and group T-joint endoscopy mask.Topical anesthesia of nasal cavity,pharynx,larynx and trachea with 2％ lidocaine was conducted and then remifentanil was continuously infused at 0.05-0.15 μg· kg-1 · min-1 in both groups.The incidence of hypoxemia and intubation time were recorded.Arterial blood samples were obtained for determination of PaO2 and PaCO2 before topical anesthesia (baseline),immediately before and 1 min after placement of FOB and immediately after nasotracheal intubation was accomplished.Results The incidence of hypoxemia was significantly lower in group Tjoint endoscopy mask (0) than in group nasal catheter (25％) (P ＜ 0.05).The PaO2 during nasotracheal intubation was significantly higher in group T-joint endoscopy mask than in group nasal catheter (P ＜ 0.05).There was no significant difference in PaCO2 and intubation time between the 2 groups (P ＞ 0.05).Conclusion T-joint endoscopy mask facilitates awake nasotracheal intubation without affecting oxygen inhalation in patients with cervical spinal cord injuries.

Objective To evaluate the efficacy of an airway topical anesthesia catheter for topical anesthesia using a spray-as-you-go technique via the fiberoptic bronchoscope (FOB).Methods Forty American Society of Anesthesiologists physical status Ⅰ-Ⅲ patients with obstructive sleep apnea syndrome,aged 20-64 yr,with body mass index of 23-35 kg/m2,with no upper respiratory tract infection within 1 week before operation,scheduled for elective uvulopalatopharyngoplasty,were divided into 2 groups (n =20 each) using a random number table:routine control group (group C) and FOB-airway topical anesthesia catheter group (group F).In group C,the pharynx and larynx were sprayed with lidocaine FOB by using a laryngo-tracheal mucosal atomization device,and cricothyroid membrane puncture was performed and then lidocaine was injected.In group F,airway topical anesthesia was performed using a spray-as-you-go technique via the FOB with an airway topical anesthesia catheter spraying lidocaine via the nose.At 5 min after topical anesthesia of the airway,FOB-guided intubation was performed,and dexmedetomidine was intravenously infused at 0.1 μg · kg-1 · min-1 for sedation in both groups.Ramsay sedation scores were assessed after topical anesthesia and before intubation.The scores for the intubating condition and tolerance of tracheal tube were assessed during FOB-guided intubation.Successful intubation and the development of responses to intubation and hypoxemia were recorded.The patients were followed up one day after the end of operation,and parents' satisfaction with the procedure of intubation was recorded.Results Compared with group C,the intubating condition score,tolerance of tracheal tube score,success rate of intubation at first attempt and rate of parents' satisfaction with the procedure of intubation were significantly increased,and the incidence of responses to intubation was decreased (P＜0.05),and no significant change was found in Ramsay sedation scores before intubation and incidence of hyoxemia in group F (P＞0.05).Conclusion When the FOB is used to guide awake nasotracheal intubation,the airway topical anesthesia catheter provides better efficacy,better intubating conditions,and fewer side effects when applied for topical anesthesia using a spray-as-you-go technique via the FOB,it can be easily accepted by the patients and the efficacy is better that of routine airway topical anesthesia.

Objective To investgate the changes in the expression of hepatocyte growth factor (HGF)and c-met in the lungs in a rat model of pulmonary hypertension.Methods Eighty 7 week old male SD rats weighing 180-250 g were randomly divided into 2 groups ( n =40 each ):control group (group C) and pulmonary hypertension group (group PH).Pulmonary hypertension was induced by left pneumonectomy and subcutaneous monocrotaline (MCT) 60 mg/kg 2 weeks later.Pulmonary artery pressure and the ratio between the weight of right ventricle and left ventricle + interventricular septum ( RV/LV + S) were measured at 7,14,21 and 28 d after MCT administration.HGF and c-met protein and mRNA expression and TGF-β content in the lung tissue were determined.Results Pulmonary hypertension and right ventricular hypertrophy associated with hypertrophy of pulmonary artery tunica media and muscularization of small pulmonary arteries developed after MCT administration in PH group.In PH group HGF protein and mRNA expression in the lungs was significantly down-regulated as compared with group C.There were no significant differences in c-met protein and mRNA expression in the lungs between the 2 groups.The TGF-β content in the lungs was significantly increased in group PH as compared with group C.Conclusion Decrease in HGF production in the lungs plays an important role in the pulmonary hypertension.Increasing of pulmonary TGF-β may play an important role in the down-regulation of pulmonary HGF expression during pulmonary hypertension.

Objective To investigate the effect of transplantation of bone marrow mesenchymal stem cells (MSCs) genetically modified with human hepatocyte growth factor gene (hHGF) on angiogenesis in the rat lung.Methods Twenty F344 rats,aged 2 months,weighing 200-250 g,were randomly divided into 2 groups ( n =10 each):HGF group and control group (group C).MSCs genetically modified with hHGF was injected through the external jugular vein in group HGF.While the equal volume of DMEM culture medium (1 ml) was given instead in group C.The mean pulmonary artery pressure was detected at 28 days after transplantation.Then the rats were sacrificed and the lungs were removed for determination of the content of hHGF,expression of proliferating cell nuclear antigen (to reflect the degree of endothelial cell proliferation showed by the small pulmonary vessels) and Ⅷ factor (to reflect the density of the small pulmonary vessels),and microscopic examination.Results Compared with group C,no significant change was found in mean pulmonary artery pressure ( P ＞ 0.05),while the content of hHGF,degree of endothelial cell proliferation,and density of the small pulmonary vessels were significantly increased in group HGF ( P ＜ 0.01).No change was found in the structure of the small pulmonary vessels in group HGF.Conclusion Transplantation of MSCs genetically modified with hHGF can promote angiogenesis in the rat lung.

Objective To investigate the effect of human hepatocyte growth factor (hHGF) genetic modification on the ameliorating effects of mesenchymal stem cells (MSCs) implantation on pulmonary microvascular rarefaction in a rat model of pulmonary hypertension (PH).Methods MSCs were obtained from F344 rats and transduced with lentiviral vector modified with human HGF (hHGF-MSCs) or empty vector (EGFP-MSCs).Sixty-six 7 week old male F344 rats weighing 180-250 g were used in this study.PH was induced by left pneumonectomy and subcutaneous monocrotaline (MCT) 60 mg/kg injected at 2 weeks after operation.The animals with PH were randomly divided into 3 groups:control group (group C),EGFP-MSCs group (group E) and HGF-MSCs group (group H).Groups H and E received hHGF-MSCs or EGFP-MSCs 5 × 105 in DMEM 1 ml iv at 3 weeks after subcutaneous MCT injection,while group C received plain DMEM 1 ml.Mean pulmonary arterial pressure (mPAP) was measured and right ventricular hypertrophy and angiogenesis in the lung were assessed and the content of rat HGF (rHGF) and hHGF protein in lung tissue and pulmonary capillary density (by immuno-histochemistry) was measured at 2 weeks after MSCs implantation.The survival rates within 45 days after MCT administration were compared among the 3 groups.Results No hHGF was detected in groups C and E.Both hHGF-MSCs and EGFP-MSCs significantly reduced MPAP and right ventricular hypertrophy and increased pulmonary capillary density and survival rates in groups H and E as compared with group C and the efficacy of hHGF-MSCs was significantly greater than that of EGFP-MSCs.Barium angiography revealed that distal pulmonary vasculature was significantly increased in group H as compared with groups E and C.The survival of the rats receiving hHGF-MSCs was significantly longer in group H than that in groups E and C.Conclusion hHGF genetic modification can improve the ameliorating effects of MSCs implantation on PH-related microvascular rarefaction.

Objective To construct F344 rat bone marrow mesenchymal stem cell line (MSC) modified with human hepatocyte growth factor (hHGF) gene.Methods Recombinant virus containing hHGF was obtained by transfecting the packaging cell line 293 FT with lentiviral vector pLV/EF1α-hHGF-IRES-eGFP.MSCs derived from F344 rat bone marrow were then tranfected with packed lentiviral vector.Purified MSCs expressing hHGF was obtained by screening culture with G418.MSCs and MSCs transfected with empty vector were used as control.The expression of hHGF protein was detected by Western blot (eGFP-MSCs).The hHGF-transfected MSCs were cultured in osteoblast-inducing culture medium and osteoblast phenotype was assayed by alizarin Red staining.The cells were also cultured in adipogenesis medium and stained with Oil Red O for identification.Results The expression of hHGF protein was significantly up-regulated in the hHGF-MSCs as compared with MSCs and eGFP-MSCs.hHGF-MSCs readily differentiated into mineralizing cells or adipocytes when incubated in differentiation medium.Conclusion A F344 rat MSC line that stably expresses HGF is successfully established.

Objective:To explore the early diagnosis and correct treatment of neurogenic pulmonary edema (NPE) and review the literature.Method:Retrospective analysis was performed in six patients diagnosed as NPE who were admitted to the emergency department of Tianjin Third Central Hospital from March 2017 to March 2021.Results:Six patients had acute onset, presenting severe dyspnea and hypoxemia, and obvious wet rales could be heard in both lungs. The white blood cell count (WBC) increased to varying degrees (11-22)×10 9/L, procalcitonin (PCT) was normal, or slightly increased, sputum bacteriological examination was negative, and oxygenation index was < 200 mmHg (1 mmHg≈0.133 kPa). Chest CT mainly showed patchy or patchy exudation. The lesions were of different sizes and were not distributed according to lobes. By reducing intracranial pressure, ventilator assisted breathing, liquid therapy, anti-infection therapy with antibiotics, nutritional support, all six patients were well and discharged, and no one died of NPE. Conclusions:NPE has complex condition, acute onset and rapid development. Early diagnosis and correct treatment can improve the success rate of treatment and prognosis of patients with NPE.

Objective To investigate the status and needs of training medical staff in county-level medical institutions in Guangxi to provide preferences for developing training programs. Methods Self-designed questionnaires were used to investigate a random sample of 477 medical staffs in 9 county-level medical institutions including People's Hospi-tals, Chinese Medical Hospitals, Maternity and Child Care Hospitals in Wuming, Liujiang, Rongxian counties. Re-sults 13% of the medical staffs had never participated in training; 28.1% of them were trained once a month; 50.7% of them were trained for less than a week. 39.4% of them expected to receive training once a year , 21.6% expected a week to half a month and 21.8% expected three months to six months. 60.4% of them expected to receive new knowl-edge and technique training. 73.6%of them expected to go to provincial hospitals for training and 57% of them expect-ed to receive extension study. Conclusion There are differences between medical staff training needs and status. Exist-ing questions are incomplete training coverage, too dense training frequency ,too short training period, medical staffs urgently need the training of continuing professional development, new technology and professional knowledge, and training effect is not obvious. We can establish medical staff in-service training incentives ,increase medical staffing and reform training methods to encourage and attract more medical staff to attend training.