The etiological agent of severe acute respiratory syndrome (SARS) was identified as a new coronavirus, termed SARS-CoV. Establishment of an efficient and sensitive diagnostic system of SARS-CoV genetic materials is crucial for SARS control. In this study, we quantified SARS-CoV mRNAs in both infected cell culture lysate and in supernatant by using Real-time quantitative revere transcription-PCR based on EvaGreen鈩?dye and Taqman-MGB probes. For extensive evaluation of sensitivities and specificities, 13 pairs of primers and 4 probes were designed based on different genes of SARS-CoV. Glyceraldehydes-3-phosphate dehydrogenase (GAPDH) was selected as the internal control gene. Results showed that S-gene-specific PCR was the most sensitive for detection, but because of its sequence variability in the different viral strains, primers and a probe based on the N gene were suitable substitutions. Meanwhile, we found the mRNA concentrations in cell culture lysates were much higher than in cell supernatant and facilited more sensitive detection of the SARS-CoV.

Objective To study their chemical components of xianglingcao(Crotalaria ferruginea) and identify their chemical structures.Methods The compounds were isolated by chromatography and their structures were identified by spectral analysis and compared with the published data.Results Ten compounds were isolated and identified as vomifolilol(Ⅰ),genistein(Ⅱ),p-hydroxybenzoic acid(Ⅲ),5,7-dihydroxy-4-methoxy-flavone-7-O-?-D-glycopyranoside(Ⅳ),octadecoic acid(Ⅴ),octacosanol(Ⅵ),?-stitosterol(Ⅶ),?-daucosterol(Ⅷ),stigmasterol(Ⅸ),and ?5,22 stigmasterol-3-O-?-D-glycopyranoside(Ⅹ).Conclusion The ten compounds are obtained from xianglingcao for the first time.

Objective: To study the incidence of ventricular arrhythmia (VT) in heart failure (HF) patients after cardiac resynchronization therapy (CRT-D) and identify the influencing factors for VT occurrence, to explore the impact of CRT-D shocks on mortality and the management of appropriate shocks. Methods: A total of 42 patients with successfully implanted CRT-D in our hospital from 2009-01 to 2015-04 were studied. There were 2 groups of patients: Ischemic cardiomyopathy group,n=12 including 8 patients for primary prevention and 4 for secondary prevention; Non-ischemic cardiomyopathy group,n=30 including 19 patients for primary prevention and 11 for secondary prevention. The patients with appropriate shocks received four step-wise therapy as drug, equipment parameter adjustments, revascularization and radiofrequency ablation (RA). Results: The patients in Ischemic cardiomyopathy group were followed-up for (38.1±24.0) months, 7 patients suffered from post-operative VT, 5 patients had CRT-D appropriate shocks. The patients in Non-ischemic cardiomyopathy group were followed-up for (27.5±17.8) months, 11 patients suffered from post-operative VT, 10 patients had CRT-D appropriate shocks. The occurrence rates of post-operative VT and CRT-D appropriate shocks were similar between 2 groups,P>0.05; the success rate for ATP treating VT was higher in Ischemic cardiomyopathy group (69% vs 55%),P<0.05. Cox regression analysis indicated that CRT-D as secondary prevention was the independent influencing factor for VT occurrence,P=0.001. During follow-up period, 9 patients with shocks died; the mortality in patients with shocks was higher than those without shocks (43% vs 0%),P<0.05. With four step-wise therapy, 80% patients in Ischemic cardiomyopathy group had no more shocks; with three step-wise therapy as drug, equipment parameter adjustments and RA, 90% patients in Non-ischemic cardiomyopathy group had no more shocks, 10% patients had reduced shocks. Conclusion: CRT-D as secondary prevention was the independent impact factor for post-operative VT occurrence, no matter appropriate or inappropriate shocks would elevate the risk of death in HF patients. Step-wise therapy was important to reduce appropriate shocks.

To study the effects of rCBF and brain function in the patients with cerebral infarction byearly rehabilitation training. 89 cases were randomized into rehabilitation and control groups and were ex-amined rCBF by 133Xe inhalation method and BEAM. Total effect rate was 93.9%in rehabilitationgroup,to the control 77.5%(X2=3. 95,P<0.05). The rCBF rised up in two groups,but it was higher inthe foriner,to the contro1,t=4. 99,P<0. 01. BEAM improve rate was 95.9%,to the control,77.5%(X2=5. 30,P<0. 05). So we confirmed that early rehabilitation training may promote rCBF and improve brainfunction of patients with cerebral infarction.

Objective This study compares a dual-freeze protocol with a triple freeze protocol for hepatic cryoablation in the Tibetan pig model.Method Cryoablation with a dual-(10-5-10-5 min)and triple-freeze (5-5-5-5-10-5 min) protocol for the normal livers of 9 Tibet pigs was performed under exposed operation.Temperature changes of cryoprobes and diameter changes of iceballs were measured during the ablation,and seven days later the pathological changes of cryozones were reviewed and the surface and depth cryolesions were measured.Results Compared with cryoablation with two freeze-thaw cycles,there was a greater iceball diameter for cryoablation by three freeze-thaw cycles.Also,seven days after cryosurgery,there were similar surface and deep cryolesions in dual-and triple-freeze protocols.Pathologically,the triple freezing protocol was associated with a longer zone of complete necrosis.Conclusions With the same freezing time (20 min),the triple-freeze protocol may become a more powerful liver-ablation method in cryosurgical application.

AIMTo investigate the transformation of characteristics of epidermal cells from foreskin which were used to reconstruct male rabbit anterior urethra in combination with acellular collagen matrices.

METHODSIn three rabbits, autologous foreskin epidermal cells were isolated, expanded in vitro, and seeded (inoculated) onto a tubular acellular collagen matrix, acquired from allogeneic rabbit bladder submucosa. A urethral mucosal defect was created, and urethral reconstruction was performed with the tubular acellular collagen matrix seeded with epidermal cells.

RESULTSOn gross examination at 12 months following the procedure, the mucosa of the urethral grafts appeared lubricous and smooth. Urethrography showed that a wide urethral caliber had been maintained without any sign of strictures. Histological examination showed a transitional cell layer in the graft without evidence of a margin between the graft and the host tissue at 12 months postoperatively.

CONCLUSIONEpidermal cells seeded onto acellular collagen matrices can be successfully used to reconstruct urethras that have defects and are transformed to transitional epithelial cells.

Animals ; Cell Transplantation ; methods ; Collagen ; Epidermis ; cytology ; Foreskin ; cytology ; Graft Survival ; Male ; Mucous Membrane ; cytology ; Rabbits ; Reconstructive Surgical Procedures ; methods ; Tissue Culture Techniques ; Tissue Engineering ; methods ; Urethra ; surgery ; Urethral Stricture ; surgery

Objective:To observe the clinical efficacy of Ziyin-Heyang Decoction on the treatment of glucocorticoid-dependent primary immune thrombocytopenia. Methods:A total of 70 patients with ITP in Xiyuan Hospital of China Academy of Chinese Medical Sciences from November 2015 to November 2017 who met the inclusion criteria were divided into two groups according to the random number table method. 33 cases in the treatment group and 32 cases in the control group completed the reaserch. The control group received standard dose of prednisone acetate orally, and the dose was gradually reduced according to platelet count; and the treatment group was given Ziyin-Heyang Decoction on the basis of the control group. Both groups had one course of treatment for 8 weeks, with a total of 2 courses of treatment. Before and after treatment, TCM syndrome score was performed, PLT was detected by impedance method, bleeding score was calculated, daily dosage of prednisone acetate tablets was recorded, and platelet related antibodies (PAIgA, PAIgG, PAIgM, PAIgD) were detected by flow cytometry. Then clinical and syndrome efficacy were evaluated. Results:The total effective rate was 75.8% (25/33) in the treatment group and 53.1% (17/32) in the control group, and the difference in efficacy was statistically significant ( Z=2.846, P<0.05). For the TCM syndromes, the total effective rate was 100.0% (33/33) in the treatment group and 75.0% (24/32) in the control group, and there was a statistical difference between them ( Z=3.841, P<0.05). After treatment, the platelet count improvement [(97.6 ± 39.9)×10 9/L vs. (70.9 ± 16.6)×10 9/L, t=3.540] of the treatment group was significantly higher than that of the control group ( P<0.05). The TCM syndrome score [6.00 (4.00) vs. 10.00 (6.00), Z=3.510] of the treatment group was significantly lower than that of the control group ( P<0.05). After the treatment, the dose of prednisone acetate tablets [5.00 (17.50) mg/d vs. 27.66 (11.98) mg/d, Z=4.186] of the treatment group was significantly lower than that of the control group ( P<0.05). Bleeding score decreased in both groups, but there was no significant difference between them ( P>0.05). There was no statistically significant change in platelet related antibody levels in the treatment group before and after treatment ( P>0.05). Conclusions:The Ziyin-Heyang Decoction has advantages in improving platelet count, with drawing glucocorticoid and improving symptoms.

Objective To explore the clinical significance of prenatal diagnosis for absence of fetal cavum septum pellucidum (CSP) by ultrasonography. Methods The ultrasonographic characteristics were retrospectively analyzed in 63 fetuses, whose CSPs were not detected in prenatal two and three dimensional ultrasonography in Nanjing Medical University Affiliated Suzhou Hospital. Results In 63 fetuses with absent CSP, the related malformations included:(1) Five cases were diagnosed as agenesis of corpus callosum (ACC, including four complete ACC and one partial ACC);(2) Twenty-seven cases were diagnosed as holoprosencephaly (HPE, including 18 alobar HPE, ifve semilobar HPE and four lobar HPE);(3) One case was diagnosed as schizencephaly;(4) Two cases were diagnosed as porencephaly;(5) Five cases were diagnosed as hydranencephaly;(6) Twenty-three cases were diagnosed as severe hydrocephalus, among which ifve cases were caused by aqueductal stenosis and the other 18 cases were caused by open spina biifda (Chiari Ⅱ malformation). These absent CSP fetuses had other abnormalities including one case of Dandy-Walker Malformation, one case of Dandy-Walker Variation, 14 cases of median cleft lip, ifve cases of single naris, two cases of proboscis nose, three cases of talipes foot and four cases of single umbilical artery, etc. These 63 cases were all followed up after prenatal ultrasonography but 4 cases were missed and one case of septo-optic dysplasia was misdiagnosed as lobar HPE. The diagnoses of other 58 fetuses by ultrasonography were conifrmed by follow up. Conclusions CSP is considered as an essential part in prenatal evaluations of the fetal central nervous system during second-and third-trimester. Absence of CSP is always accompanied with an extremely wide spectrum of fetal neuroanatomic malformations, including prosencephalon diseases, intracranial midline anomalies, etc. Prenatal ultrasonography is the best choice for the observation of CSP.

Objective To investigate the prevalence and incidence of Keshan disease (KD) and the selenium concentration of food and hair in residents of Yongjin Village, Fuyu County, Heilongjiang Province, national monitoring site, in 2007. Methods According to the Standard of Keshan Disease Surveillance and the Standard of Diagnosis of Keshan Disease(GB 17021-1997), the residents living in the monitoring site were surveyed by clinical examination and electrocardiography. For individuals whose hearts showed abnormalities, a chest X-ray photograph was taken. The selenium concentrations of the residents' food (flour) and hair were assayed by flowing injection hydride generation atomic fluoremetric method(FI-HG-AFM). Results Nineteen KD patients were found from 282 residents in 2007 KD surveillance. The prevalence of KD, latent KD and chronic KD were 6.7%(19/282), 2.8%(8/282) and 3.9%(11/282), respectively. Five of the 8 latent KD cases were newly found. In addition, there were 5 the suspected KD cases, including 2 suspected chronic KD cases. No acute KD or sub-acute KD patients were found in Yongjin Village at this monitoring site this year. The average selenium concentration of children hair and residents food were (0.3197±0.0586)mg/kg and (0.0210±0.0062)mg/kg, respectively. Conclusions New cases of KD continued to emerge, indicating that etiological factors still exist. Therefore, the emphasis of monitoring KD in furore is founding the consummate report of infectious disease system and training the personnel to increase the reliability of monitoring.