AIM: To apply UASLG for Rana temporatia chensinensis David's identification. METHODS: Rana temporatia chensinensis David was made ultrasonic extraction. These solvents in turn were petroleum ether、chlorofo mn、ethyl acetate、n-butanol and 95%WTBZ# ethanol,then their absorbance in UV and one-order derivative spectrums were determined. RESULTS: Rana temporatia chensinensis David's UASLG was reproducible. CONCLUSION: UASLG's action spectrums for Rana temporatia chensinensis David can be identified for quality control.

The author proposed student-centered learning system in otolaryngology-head and neck surgery for undergraduate clinical training after exploration and intentions.Four mutual impacted frames were built including integration of teaching philosophy,visualization of training methods,diversification of educational targets and interaction of training courses.Endoscopic navigated learning and multimedia aided training were applied,respective teaching purposes were set and various clinical training courses were introduced to students in their learning of otolaryngology,which were believed to help develop more medical talents with higher comprehensive qualities and better clinical skills.

Gastric cancer carcinogenesis is a multifactorial process which is related to an interaction of host factors,Helieobacter pylori (Hp) infection,dietary factors and so on.The plant polyphenols which are widely present in many plants are the general term for a large group polyphenolic compounds.They have strong antioxidant activity.Furthermore,many animal experiments and clinical studies have proved that the polyphenols could inhibit gastric cancer via inhibiting Hp infection,suppressing the expression of nuclear factor-κB (NF-κB),promoting apoptosis in cancer cells and so on.The application of plant polyphenols could broaden the approaches for chemoprevention of gastric cancer.

OBJECTIVE: To explore the treatment and possible prognostic factor of nasal sinuses adenoid cystic carcinoma. METHOD: Retrospectively analysed the records of 18 patients with complete clinical and pathological data,which including 4 patients given up treatment, 5 patients taken surgical treatment and 9 patients taken surgical treatment as well as radiotherapy and chemotherapy. RESULT: Fifty percent of the patients got 2-year survival and 3 cases of death due to intracranial tumor invasion and 2 patients died of the disease distant metastasis. CONCLUSION If patients got Nasal sinuses adenoid cystic carcinoma, they should take comprehensive treatment based on surgery, in order to improve the survival rate. The prognosis depends on the tumor early detection and early treatment, the sooner the treatment, the better.
Adult ; Aged ; Carcinoma, Adenoid Cystic ; therapy ; Female ; Humans ; Male ; Middle Aged ; Nasal Cavity ; pathology ; Paranasal Sinus Neoplasms ; therapy ; Prognosis ; Retrospective Studies

Visceral myopathy is a rare disorder characterized by atrophy,losing and fibrosis of visceral smooth muscle.Digestive tract is often invaded by the lesions,and the symptoms are various according to different lesions and degrees.Small intestinal involvement is characterized by abdominal distension,diarrhea and vomiting.Colon involvement is characterized by chronic intestinal pseudoobstruction.Malnutrition and hypoproteinemia may be secondary to this disease.The diagnosis of visceral myopathy is difficult,paralytic ileus,chronic constipation and systemic sclerosis must be considered in the differential diagnosis.The progression of the disease is slow,and the longterm prognosis is poor.In this article,the diagnosis and treatment of visceral myopathy causing acute intestinal pseudoobstruction were introduced based on the clinical data of 1 patient.

BACKGROUND: There still was not any report about inducing stem cells into matured cells to form products in vitro.OBJECTIVE: To induce CD34+ cells of umbilical cord blood to differentiate into mature megakaryocytes, and to investigate the mechanism of production of platelets.DESIGN, TIME AND SETTING: This cytology in vitro study was conducted at the Central Laboratory of Xiangya Hospital and Xiangya Third Hospital from 2004 to 2006. MATERIALS: Umbilical cord was collected from healthy full-term pregnant puerperants at the Xiangya Hospital.METHODS: The CD34+ cells were isolated from umbilical cord blood by magnetic activated cell sorting (MACS) and then cultured in 24-well culture plate at 5x107/L in StemPro-34 serum-free medium, supplemented with L-glutamine, saturated human transferrin, CaCl2, insulin, deionized bovine serum albumin and recombinant human thrombopoietin at 37℃, under 0.05 volume fraction CO2 saturated humidity to be differentiated into megakaryocytes for 14-21 days. Cell medium was absorbed, and centrifuged to obtain supernatant. Samples were centrifuged again, and then supernatant was removed. The remaining was platelet-like particles in cell culture plate. Platelet was isolated from normal platelet-rich plasma.MAIN OUTCOME MEASURES: The following parameters were measured: morphological changes in cultured cells and platelet-like particles in supematant; results of immunohistochemistry; observation results under a microscope; platelet aggregation; CD41 expression.RESULTS: At day 10, silk-like substances were found in megakaryocyte culture medium, with the presence of platelet-sized particles. The production of platelet-sized particles reached a peal at day 16. Cultured cells were strongly positively for platelet-specific antigen GP Ⅱb Ⅲa. Under the optical microscope, mature megakaryocytes were detected, with the presence of some immature megakaryocytes, and platelet-sized particles were found surrounding megakaryocytes. Under the electron microscope, a majority of mature megakaryocytes and a few apoptotic megakaryocytes were detected, and platelet-sized particles in the supernatant had the same size and structure with the platelet in the platelet-rich plasma. Some platelet surfaces were smooth or irregular. Platelet-sized particles in the supematant aggregated in response to thrombin as platelets in normal platelet-rich plasma. Flow cytometry demonstrated that the cultured platelets had the same high expression rate of CD41 as the platelets from platelet rich plasma.CONCLUSION: Umbilical cord blood CD34+ cells can be induced to differentiate into pudfied and mature megakaryocytes and platelets in vitro.

Objective To investigate whether HSP90α could be a sensitive and specific serum biomarker for the diagnosis and progression of lung cancer. Methods In the present study, different secretomic analy-ses on the two human lung adenocarcinoma cell lines CL1-0 and CL1-5 with low and high metastatic poten-tial, respectively, were performed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ma-trix-assisted laser desorption/ionization time-of-flight mass spectrometry. The candidate biomarker was con-firmed by Western blotting, and was further analyzed in 224 serum samples including 141 lung cancer, 37 benign pulmonary diseases, as well as 46 healthy individuals using ELISA assay. Results HSP90α was sig-nificantly upregulated in the CM of CL1-5 cells. It was found that the levels of HSP90α were specifically ele-vated in the sera of non-small cell lung cancer compared with other groups. At the cut-off point 0.535 on the receiver operating oharacteristie curve, HSP90α could comparatively discriminate lung cancer from benign lung disease and healthy control groups with sensitivity of 0. 817, specificity 0. 919 and total accuracy 80. 14%. Conclusion HSP90α may be a potential useful serum biomarker for discriminating lung cancer from benign lung diseases and healthy individuals and staging of non-small cell lung cancer.

Objective To evaluate the effects of lentivirus-delivered short hairpin RNA (shRNA) targeting human papillomavirus 16 (HPV16) E7 gene on the expression of 4 kinds of DNA methyltransferases (DNMTs),including DNMT1,DNMT3A,DNMT3B and DNMT3L,in HPV16-positive cervical cancer cell line SiHa.Methods The recombinant plasmid containing HPV16 E7 gene-targeting shRNA was constructed firstly.Then,the BLOCK-iTTM lentiviral RNAi expression system kit was used to package the lentiviral vector,which was transfected into 293T cells.The lentivirus-containing supernatants were collected at 48 and 72 hours after transfection.The SiHa cells were divided into 3 groups to be cultured with lentiviral supernatant containing HPV16 E7 gene-targeting shRNA recombinant plasmids mixed with complete medium at a ratio of 1:1 (shRNA group),lentiviral supernatant containing empty plasmids mixed with complete medium at a ratio of 1:1 (negative control group),and complete medium alone (blank control group),respectively.Real-time fluorescence-based quantitative PCR (qRT-PCR) was performed to measure mRNA expression of HPV16 E7 and 4 kinds of DNMTs in the above 3 groups at 0,48,96 hours after infection,and Western blot analysis to determine protein expression of the 4 DNMTs at 48,96 hours after infection.Results There were no significant differences in the mRNA expression of HPV16 E7 and the 4 DNMTs among the shRNA group,negative control group and blank control group at 0 hour after infection (all P ＞ 0.05).At 48,96 hours after infection,the mRNA expression of HPV16 E7 and the 4 DNMTs decreased significantly in the shRNA group compared with the negative control group and blank control group (all P ＜ 0.05),but did not differ between the negative control group and blank control group (all P ＞ 0.05).Additionally,E7,DNMT1,DNMT3A,DNMT3B and DNMT3L gene-silencing efficiencies in the shRNA group were 71.13％,50.53％,13.72％,46.27％ and 17.92％ at 48 hours,and 83.50％,74.2％,47.8％,64.7％ and 48.9％ at 96 hours after infection,respectively.Western blot analysis showed that the protein expression of the 4 DNMTs significantly decreased in the shRNA group compared with the negative control group and blank control group at 48,96 hours after infection (all P ＜ 0.01).Moreover,the protein expression of DNMT1,DNMT3A,DNMT3B and DNMT3L in the shRNA group gradually decreased over time,and was inhibited by 84％,37.2％,59.8％ and 49.3％ at 48 hours respectively,and by 73.1％,68.7％,55.5％ and 65.5％ at 96 hours after infection respectively.Conclusion Targeted silencing of E7 gene in HPV16-positive SiHa cells can interfere with the mRNA and protein expression of DNMT1,DNMT3A,DNMT3B and DNMT3L.

ObjectiveTo compare the predictive value of anatomic scoring system, physiological scoring system, and the combination of two systems in death prediction of patients with severe trauma in intensive care unit (ICU). Methods A retrospective analysis of patients with severe trauma admitted to department of critical care medicine of Daping Hospital, the Third Military Medical University, and Zunyi Medical University from January 2011 to December 2014 was conducted. The patients meeting the following criteria were enrolled: over 16 years old, admitted to hospital shorter than 24 hours after trauma, length of ICU stay≥48 hours, and injury severity score (ISS)≥16. Patients were divided into two groups: survivors and non-survivors. The data of anatomic scoring system, including ISS and new injury severity score (NISS), and physiological scoring system, including acute physiology and chronic health evaluationⅡ(APACHEⅡ) score were collected. The predictive power for death of the scoring system alone or combination in patients with severe trauma was evaluated.Results A total of 614 patients with severe trauma were enrolled, and there were 153 deaths with a mortality rate of 24.9%. ISS, NISS, APACHEⅡ, ISS+ APACHEⅡ, NISS+ APACHEⅡ of non-survivors were significantly higher than those of survivors (ISS: 29.15±7.75 vs. 24.31±6.50, NISS: 41.96±12.01 vs. 29.64±8.19, APACHEⅡ: 23.71±6.58 vs. 17.02±5.49, ISS+ APACHEⅡ: 52.86±10.00 vs. 41.33±8.70, NISS+ APACHEⅡ: 65.67±13.46 vs. 46.66±10.43, allP< 0.01). The area under receiver operating characteristic curve (AUC) of ISS, NISS, APACHEⅡ, ISS+ APACHEⅡ, NISS+ APACHEⅡ was 0.687, 0.792, 0.782, 0.809, and 0.860, respectively. Both of ISS+ APACHEⅡ and NISS+ APACHEⅡ had higher AUC than that of ISS, NISS or APACHEⅡ alone; and the AUC of NISS+ APACHEⅡ was significantly larger than that of ISS+ APACHEⅡ(allP< 0.05). NISS+ APACHEⅡ showed the largest AUC in death prediction of severe trauma patients. The cut-off value, sensitivity, specificity, positive predict value (+PV), negative predict value (-PV), positive likelihood ratio (+LR), negative likelihood ratio (-LR), and Youden index of NISS+ APACHEⅡ, which had the greatest AUC, were 56, 75.2%, 82.0%, 58.1%, 90.9%, 4.17, 0.30, and 0.572, respectively.Conclusion The combination of anatomic scoring system and physiological scoring system is better than single scoring system for death prediction in patients with severe trauma in ICU, and it may be considered to be a new method for early identification of death risk in patients with severe trauma.

Objective:To investigate the promotive effect of 4,5,6,7-tetrabromobenzotriazole (TBB) on the apoptosis of the human cervical cancer HeLa cells,and to clarify its effect mechanism in related signaling pathways.Methods:The human cervical cancer HeLa cells at logarithmic growth phase were divided into control group(without TBB) and experiment group(with TBB).MTT assay was used to detect the survival rate of the HeLa cells;the morphology of HeLa cells was observed under inverted microscope;Annexin Ⅴ-FITC/PI double staining and flow cytometry(FCM) were used to determine the apoptotic rates of the HeLa cells;the expression levels of apoptosis-related proteins p-Akt,Akt,Bcl-2,Bax,cleaved-caspase-3,and Pro-caspase-3 were detected by Western blotting method.Results:The MTT results showed that the survival rates of the HeLa cells in experiment group were significantly decreased(P<0.01) in a dose-dependent manner compared with control group.The apoptotic morphology of the HeLa cells in experiment group were found as cell shrinkage and karyopyknosis under inverted microscope.The Annexin Ⅴ-FITC/PI double staining and FCM results showed that the apoptotic rates of the HeLa cells in experiment group (3,6,12 and 24 h) were higher than that in control group (P<0.05 or P<0.01).The Western blotting results showed that compared with control group,the expression levels of the anti-apoptotic proteins p-Akt and Bcl-2 in HeLa cells in experiment group were decreased obviously,whereas the expression levels of the pro-apoptotic proteins Bax and cleaved-caspase-3 were increased and the expression levels of Pro-caspase-3 were decreased.Conclusion:TBB may promote the apoptosis of human cervical cancer HeLa cells by inhibiting the Akt signaling pathway.