This study aims to analyse the value of CODEHOP RT-PCR in the detection of Flavivirus. According to the amino acid sequences of polyproteins of different flaviviruses published in GenBank, a pair of primers was designed using the CODEHOP method. One-step RT-PCR was used to detect Japanese encephalitis virus strain JEV1201, Dengue virus strain JKD001, and yellow fever virus vaccine YV6161. BLAST analysis and phylogenetic analysis were performed after the RT-PCR products of nucleocapsid genes were sequenced. The results showed that this method could amplify Flavivirus specifically, and the size and sequence of the target fragment accorded with the anticipated result. JEV1201 had the highest homology to Japanese encephalitis virus strain YL2009-4/YC2009-3, belonging to the branch of the phylogenetic tree of Japanese encephalitis virus strains. JKD001 had the highest homology to Dengue virus strain DENV-2/ID/1022DN/1975, belonging to the branch of the phylogenetic tree of Dengue virus strains. YV6161 had the highest homology to Yellow fever virus strain 17D, belonging to the branch of the phylogenetic tree of Yellow fever virus strains. In conclusion, the method of CODEHOP RT-PCR can be effectively used to detect, identify, and phylogenetically analyse Flavivirus.
DNA Primers ; genetics ; Flavivirus ; classification ; genetics ; isolation & purification ; Flavivirus Infections ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Nonstructural Proteins ; genetics

Objective To investigate the clinical significance in MRP1/CD_9 expression in cervical squamous cancer tissues and normal cervical tissues.Methods The expression of MRP1/CD_9 were assayed by SABC immunohistochemical methods in 53 cases of cervical cancer tissues and 13 cases of normal cervical tissues.Results Positive expression of MRP1/CD_9 was detected in 13 normal cervical tissue.MRP1/D_9 ex- pression is down-regulated in cervical carcinoma(P

Objective To investigate the distribution and antimicrobial resistance of clinical isolates of Acinetobacter baumannii isolated from our hospitalized patients from 2008 to 2013 , and analyze the correlation be-tween the medication frequency and the drug resistance .Methods The distribution and the drug susceptibility testing for Acinetobacter baumannii isolated from clinical specimens were retrospectively analyzed .The using trend of antibiotics was analyzed by calculation the DDDs .The relation-ship between the drug resistance rate and defined daily dose ( DDDs) was statistically analyzed.Results Acinetobacter baumannii varied in drug resistance,the drug resistance rates to penicillins ,cephalosporins,quino-lones,and carbapenems were high.In 2013,no antibacterial drugs is sen-sitive to Acinetobacter baumannii rate of more than 50%.The resistance rate of Acinetobacter baumannii was negatively correlated with the dosage of piperacillin/tazobactam.Conclusion The resistance rate of Acineto-bacter baumannii is high.And the isolates are multidrug resistant .

Objective To investigate the clinical characteristics of neurosyphilis presenting clinically with cerebral infarction and study its treatment and prognosis. Methods A retrospective analysis was conducted in 41 patients with neurosyphilis who presented clinically with cerebral infarction. The clinical manifestations, neurological deficit scores, activity of daily life (ADL) scores, mental scale scores, and laboratory and imaging examination results of the patients were analyzed. Results Of the 41 patients, 23 exhibited reduced intelligence, 30 had limb paralysis, 8 had limb numbness, 2 complained of dizziness, and 1 experienced epileptic attacks. Head magnetic resonance imaging (MRI) revealed the presence of brain atrophy and multiple infarction foei without obvious space-occupying effect of cerebral edema in these patients. All the patients showed positive results of serum and cerebrospinal fluid Treponema pallidum hemagglutination assay (TPHA). After treatment with high-dose penicillin, 95.1% of the patients had improvements of the clinical symptoms. Conclusion The diverse clinical manifestations of neurosyphilis often result in misdiagnosis of the disease. Patients with neurosyphilis show multiple cerebral infarction foci and cerebral atrophy without obvious space-occupying effect on MRI. Penicillin can be an effective treatment in such cases, which generally have low rate of negative TPHA within 3 months. Such patients have more favorable clinical outcomes than those with common cerebral infarction, but poor prognosis is indicated in cases with concurrent extensive cerebral infarction.

Objective:To determine whether heme oxygenase-2(HO-2) gene deletion can attenuate oxidative injury induced by free Fe2+. Methods:Stereotactic injection of 10 μl sterile FeCl2 (10 mmol/L) was made into the right striata of HO-2 knockout mice and wild-type mice. Brain edema severity was measured at 24 h. Cell viability, protein oxidation, and lipid oxidation of the basal ganglia were determined at 72 h. Western blot analysis was applied for heme oxygenase-1 (HO-1) measurement.Results: Brain water content significantly decreased in HO-2 knockout mice at 24 h compared with wild-type mice. Protein oxidation and lipid oxidation significantly decreased in HO-2 knockout mice at 72 h compared with wild-type mice, while the striatal cell viability increased significantly. HO-1 expression at baseline and 72 h was also similar to that in wild-type mice. Conclusion:These results show that HO-2 gene deletion can protect basal ganalia cells from free Fe2+ -mediated oxidative stress injury,suggesting that selective inhibition of HO-2 may have a protective effect on brain oxidative injury.

Adolescent ; Adult ; Athletic Performance ; psychology ; Back ; physiology ; Electromyography ; Exercise ; physiology ; Humans ; Imagery (Psychotherapy) ; Male ; Young Adult

OBJECTIVE: To evaluate the clinical effects and characteristics of combined transperineal and transpubic urethroplasty for patients with complex pelvic fracture urethral distraction defect (PFUDD). METHODS: We retrospectively reviewed the clinical data of 17 male patients with complex posterior PFUDD from January 2010 to December 2019. The complications included urethrorectal fistulas in 2 patients (11.8%), urethroperineal fistula in 1 patient (5.9%). Ten patients had undergone previous treatments: dilatation in 3 patients (17.6%), internal urethrotomy in 1 patient, failed urethroplasty in 6 patients (35.3%), of whom 2 patients had two times of failed urethroplasties. All the patients were performed with urethroplasty by combined transperineal and transpubic approach with removing the entire pubic bone followed by the anastomosis. RESULTS: The mean age of the patients included in this study was 35.5 (range: 21-62) years. The mean length of stricture was 5.5 (range: 4.5-7.0) cm, the mean follow-up was 27 (range: 7-110) months, the mean time of operation was 190 (range: 150-260) min, the mean evaluated blood loss was 460 (range: 200-1 200) mL. There were 5 patients who needed blood transfusion intraoperatively or postoperatively. Wound infection was seen in 4 out of 17 patients and thrombosis of lower extremities in 1 out of 17 patients. The last follow-up showed that the mean postoperative maximum urinary flow rate was 22.7 (range: 15.5-40.7) mL/s. After removing the catheter, one patient presented with decreased urinary flow and symptoms of urinary infection. Cystoscopy showed the recurrent anastomotic stricture, which was cured by internal urethrotomy. In our series, the success rate of the combined transperineal and transpubic urethroplasty was 94.1% (16/17). CONCLUSION Combined transperineal and transpubic urtheroplasty can achieve a tension free anastomosis after removing the entire wedge of pubis in some patients with complex PFUDD. However, this procedure should be completed in a regional referral hospital due to the complexity of the operation and the high percentage of complications.
Adult ; Anastomosis, Surgical ; Fractures, Bone/surgery* ; Humans ; Male ; Middle Aged ; Pelvic Bones ; Retrospective Studies ; Treatment Outcome ; Urethra ; Urethral Stricture ; Young Adult

Alveolar epithelial type II (AT II) cells are essential for lung development and remodeling, as they are precursors for type I cells and also produce other non-repair cells (fibroblasts). Progenitor cells are believed to possess capability of multi-potent transdifferentiation, which is closely related to the niche, suggesting the importance of establishment of a lung progenitor cell niche model. We hypothesized that pulmonary surfactant-associated protein A (SPA) suicide gene system would cause AT II cell to kill itself through apoptosis and leave its niche. In vitro, the recombinant adeno-associated virus vectors-SPA-thymidine kinase (rAAV-SPA-TK) system was established to get targeted apoptotic AT II cells. The apoptosis of AT II cells was detected by using MTT. The results showed that cloned SPA gene promoter had specific transcriptional activity in SPA high expression cells, and SPA high expression cells (H441) transfected with TK gene had higher sensitivity to ganciclovir (GCV) than SPA low expression cells (A549). In vivo, increased apoptosis of AT II cells induced by GCV in rAAV-SPA-TK system was observed by TUNEL. Finally, the successful packaging and application of rAAV-SPA-TK system provide experimental basis to get specific lung progenitor cell (AT II) niche in vitro and in vivo.

Teaching rounds are an important part of hospitals at all levels to carry out clinical teaching work and improve medical quality. Taken anesthesia-related knowledge as the center, based on the patient's pathophysiology and anesthesia physiological changes, our department asked the core questions such as the impact of the operation method and anesthesia plan, then students answered, and finally closed the loop after the case. This process is called problem-based learning (PBL) teaching rounds. It is found that the teaching rounds can help improve the ability of anesthesiologists to solve problems, learn independently, and learn throughout their lives, thereby improving the teaching quality of standardized residency training of anesthesiology.

OBJECTIVETo investigate whether nuclear factor kappa B (NF-kappa B) participates in the regulatory function of lymphocyte proliferation and apoptosis in bronchial asthma and whether the regulatory effect of triptolide on lymphocyte proliferation and apoptosis is conducted through NF-kappa B.

METHODSIntervention with dexamethasone, triptolide and PDTC, a NF-kappa B inhibitor, were used to treat astmatic rats respectively. Pathological examination, airway response were determined, the NF-kappa B P65 expression in lung tissue and splenic lymphocytes by immunofluorescent assay were adopted, proliferative cell nuclear antigen (PCNA) in splenic lymphocytes was measured by immunohistochemistry, apoptosis of splenic lymphocytes were monitored by flow cytometry and NF-kappa B activity was investigated by electrophoresis mobility shift assay (EMSA).

RESULTSThe nuclear expression and DNA binding activity of lung tissue and splenic lymphocytes in asthmatic rats were all significantly higher than those in the control (all P < 0.05), so was the proliferation rate of splenic lymphocytes (P < 0.05), while the apoptosis rate was much lower than that of normal control (P < 0.05). Administration of PDTC could reduce the up-regulated expression and activity of NF-kappa B, the proliferation of splenic lymphocytes lowered, while the apoptosis increased. NF-kappa B activity showed an obviously positive correlation with proliferation of splenic lymphocytes (r = 0.89, P < 0.05) and a significantly negative correlation with apoptosis rate (r = -0.54, P < 0.05). After asthmatic rats had been treated with triptolide in vivo, the NF-kappa B nuclear expression and activity in airway and splenic lymphocytes, as well as the proliferation rate of splenic lymphocytes all lowered significantly (all P < 0.05), the apoptosis rate increased significantly (P < 0.05), at the same time, the inflammatory cell infiltration and high reactivity of airway were significantly alleviated (both P < 0.05). There were obviously positive correlation between the amount of airway eosinophils and reactivity with activity of NF-kappa B (r = 0.79 and r = 0.68, P < 0.05), which indicated that the effect of triptolide was not significantly different from that of dexamethasone (P > 0.05).

CONCLUSION(1) NF-kappa B participates the formation of airway inflammation and hyper-reactivity in asthmatic rats by positive regulation on proliferation and negative regulation on apoptosis of lymphocytes. (2) Triptolide reduces airway inflammation by way of inhibiting NF-kappa B, and further inhibiting the proliferation of lymphocytes, so that to give full play of the role of anti-asthmatic airway inflammatory agents. Whether the molecular mechanism of triptolide in inhibiting NF-kappa B simulates that of glucocorticoid needs further studying.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Apoptosis ; drug effects ; Asthma ; metabolism ; pathology ; Cell Division ; drug effects ; Dexamethasone ; pharmacology ; Diterpenes ; pharmacology ; Epoxy Compounds ; Lung ; metabolism ; Lymphocytes ; pathology ; Male ; NF-kappa B ; metabolism ; Phenanthrenes ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Transcription Factor RelA