Objective To apply DNA barcoding technology for exploring its taxonomic status and differences in the molecular biology of Cricetulus barabensis in Shaanxi Province.Methods Sixty-five samples of Cricetulus barabensis were collected from Dingbian,Jingbian Counties in northern of Shaanxi and Dali County in Guanzhong plain (Dingbian 58 samples,Jingbian 2 samples,and Dali 5 samples).According to the mitochondrial cytochrome C oxidase subunit I gene (CO I) sequence,the genetic distance was calculated and Neighbor-Joining tree was constructed.Results The genetic distance between two samples (13.16,13.21) and other 56 samples of Dingbian was 9.2％-10.0％.The genetic distance between the 56 samples of Dingbian and Jingbian was less than 1％ and Dali was 7.2％-8.3％;the average intraspecific genetic distance of Jingbian and Dali was less than 1％.The Neighbor-Joining tree showed that all the Cricetulus barabensis samples from the three counties were separated into two large branches.The samples of 13.16,13.21 from Dingbian together were classified into a class and the rest of the samples into another separate branch.At the same time,other samples from Dingbian except 13.16,13.21 and Jingbian were distributed in a small branch,and Dali samples were occupied another small branch.Conclusion Using the DNA barcoding technology,we have determined three subspecies of Cricetulus barabensis in Shaanxi Province,Dingbian has two kinds and Dali has a different subspecies.

Objective To observe the effect of Interferon-α2b on HEL cells (human erythroleukemia cell line) growth, apoptosis and JAK2 V617F mutation gene expression. Methods HEL cells were placed in RPMI1640 containing 10% FBS and incubated in a cell incubator. Cells in the logarithmic growth phasem were collected, adjusting the cell density to 1 × 105/mL for experimental research. The interferon concentration in five groups were 0, 5 × 105, 10 × 105, 50 × 105, 100 × 105 U/L, with different incubation time (0, 24, 72, 120 h), respectively. The cell growth status in different groups was observed in the inverted optical microscope; MTT was used to detect the inhibition of interferon on HEL cell proliferation. Cell apoptosis was detected by flow cytometry. Fluorescence quantitative PCR was used to detect the mutation gene of JAK2 V617F expression. Results Inhibition rates of Interferon on the HEL cell proliferation in 5 × 105 U/L, 10 × 105 U/L, 50 × 105 U/L, 100 × 105 U/L groups were 18.57%, 25.10%, 42.10%, 57.00%, respectively. JAK2 V617F/GAPDH by fluorescence quantitative was 1.556, 1.213, 0.870 respectively under the concentration of interferon 100 × 105 U/L for 24, 72, 120 h. Conclusions Interferon-α2b can inhibit HEL cells proliferation and induce HEL cells apoptosis. Increasing concentration of interferon increases HEL cell apoptosis rate. Interferon can inhibit JAK2 V617F expression of HEL cells in a dose-dependent manner.

Objective To investigate the clinical value of CT lymphangiography (CTL) combined with direct lymphangiography (DLG) in primary intestinal lymphangiectasia (PIL). Methods Sixteen patients diagnosed as PIL with intestinal enteroscopy were recruited in this retrospective study. All the patients were performed DLG and CTL one week before exploratory laparotomy. Subjective assessment in DLG included weak lymphatic fluid drainage, lymphangiectasia, lymphatic reflux, fistula and thoracic outlet reflux or obstruction. While for CTL combined with DLG, the intestinal and extra-intestinal lesions were evaluated, including lymph node, edema, lymphangiectasia and abnormal distribution, fistula, and lymphangiomatosis. All the diagnosis was compared with intestinal endoscopy results. Results For DLG, 16 weak lymphatic fluid drainages, 9 lymphangiectasia, 1 fistula with abdomen and 14 thoracic outlets weak lymphatic fluid drainage or obstruction were found. For DLG combined with CTL, 16 intestinal lumens dilatation and 14 circumferential intestinal thickening were found in intestinal lesions. While for extra-intestinal lesions, the imaging features included edematous findings (12 in mesentery, 7 ascites only, 2 hydrothorax and ascites, and 3 pericardial, thoracic and abdominal effusions), abdominal lymph nodes (6 cases), lymphangiectasia and abnormal distributions (14 cases), fistulas (lymph-intestinal luminal fistula in 4 cases, and lymph-abdominal fistula in 3 cases), lymphangiomatosis (3 cases), and thoracic duct outlet dysfunction and reflux (14 cases).The number of cases diagnosed as intestinal lymphangiectasia, intestinal luminal lymph exudation and lymph fistula were 16, 10 and 6 with intestinal endoscopy, while the number were 11, 0, and 4 with CTL combination with DLG. Conclusion Combination of CTL with DLG is valuable in the diagnosis of PIL.

Objective: To investigate the treatment efficacy of radiofrequency ablation (RFA) of hepatic tumors and the relevant complications. Methods: A total of 343 patients with 778 hepatic tumors underwent ultrasound-guided RFA (582 procedures). There were 212 cases of hepatic cellular carcinoma (HCC) with 448 tumors, and the average largest diameter was 4.0 cm.Of all the patients, 63 (29.7%) were in the stage of Ⅰ-Ⅱ(UICC Systems) and 149 (70.3%) in stage of Ⅲ-Ⅳ(including 43 patients with tumor recurrence after surgical resection). There were 131 cases of metastatic liver carcinoma (MLC), with 330 metastases in the liver, the average diameter was 3.9 cm, and the liver metastases of 91 patients (69.5%) came from gastrointestinal tract. The patients were treated using the relatively standard protocol. Crucial attention must be paid to monitoring the abnormal changes in ultrasound images as well as the vital signs of the patients to find the possible hemorrhage and peripheral structure injury in time. The tumors were considered ablated successfully if no viability was found on enhanced CT within 24 hours or 1 month after RFA. The patients were followed up for 2 -62 months.Results: The ablation success rate for HCC was 95.5% (428/448 tumors), and the rate for MLC was 96.4% (318/330 tumors). The local tumor recurrence rates for HCC and MLC were 8.5 % (38/448 tumors) and 11.8 % (39/330 tumors), respectively. A total of 138 patients (40.2%) underwent repeated ablations for 2-11 times because of tumor recurrence or metastasis. The first,second and third years survival rates were 87.7% , 67.4% and 56.8% for HCC patients, 81.6%, 50.8% and 27.2% for MLC patients,respectively. The survival rate from 63 early-stage HCC patients were 92.9%,82.8% and 74.5%, respectively. The major complication rate in this study was 2.4 % (14 of 582 procedures).The complications which consisted of mechanical and thermal injuries usually occurred during or shortly after the RFA treatment. There were 5 hemorrhages, 1 colon perforation, 5 injuries of adjacent structures, 2 bile leakages and 1 skin burn. Conclusion: RFA, as a minimally invasive local treatment, has become an effective and relatively safety alternative for the patients of hepatic tumors, even of advanced live tumor, tumor recurrence, liver metastases which are unresectable or difficult to treat with traditional therapies. Knowledge about possible complications and their control may increase the treatment efficacy and help to promote the use of RFA technique.

OBJECTIVETo assess the survival of radiofrequency ablation (RFA) and investigate the prognostic factors affecting overall survival, local recurrence-free survival and disease-free survival in hepatocellular carcinoma (HCC).

METHODSA total of 192 HCC patients underwent RFA treatment in our department and were enrolled into this study. Among them, 151 patients were males and 41 were females (mean age, 59.2 years, range, 24 - 87 years old). The average tumor size was (3.9 +/- 1.3) cm (range, 1.2 - 8.0 cm). Of these 192 HCC patients, their Child-Pugh grade of A, B and C were 106, 77 and 9, respectively. According to UICC-TNM system, 57, 85, 44 and 6 patients were in stage I, II, III and IV respectively. Kaplan-Meier model and log-rank test were used in univariate analysis and COX regression model was used in multivariate analysis to identify prognostic factors for survival.

RESULTSThe 1-, 2-, 3- and 4-year overall survival were 84.9%, 69.1%, 60.4% and 52.8%, respectively. Local recurrence-free survival were 75.1%, 53.8%, 43.9% and 40.8%, respectively. Disease-free survival were 64.3%, 43.2%, 37.1% and 25.0%, respectively. The following factors were identified as independent prognostic factors for survival by multivariate model: (1) Overall survival: Child-Pugh classification, standard treatment protocol and UICC-TNM staging. (2) Local recurrence-free survival: Child-Pugh classification and UICC-TNM staging. (3) Disease-free survival: UICC-TNM staging, Child-Pugh classification and daughter lesion. Among these, both Child-Pugh classification and UICC-TNM staging were independent prognostic factors for three kinds of survivals.

CONCLUSIONSDegree of tumor progress (UICC-TNM stage, daughter lesion), treatment method (applying of standard treatment protocol) and patients' liver function are the most important factors for survival after RFA. So application of proper treatment strategy before, during and after RFA should be required to improve survival.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; mortality ; pathology ; surgery ; Catheter Ablation ; Female ; Humans ; Liver Neoplasms ; mortality ; pathology ; surgery ; Male ; Middle Aged ; Prognosis ; Survival Analysis ; Survival Rate

OBJECTIVETo determine whether erythropoietin (EPO) promotes rapid proliferation of glioma through Akt pathway.

METHODSWe detected the expression of EPO in human glioma tissues using immunohistochemistry. A nude mouse model bearing human glioma U87 cell xenograft was established and given intraperitoneal injection of EPO or saline every other day, and the tumor growth was observed. In the in vitro experiment, U87 cells were treated with PBS (control), EPO, or EPO with Akt inhibitor, and the expression of p-Akt and cyclin D1 was detected using Western blotting; the cell proliferation rate was determined using cell counting kit-8 and clone formation assay, and the cell cycle changes were analyzed with flow cytometry.

RESULTSCompared with low-grade glioma tissues, high-grade glioma tissues exhibited a significantly increased EPO expression (P=0.0002). In the tumor-bearing mice, EPO treatment significantly increased the expression of EPO (P=0.0006) and p-Akt (P=0.0003) in the tumor and obviously increased the tumor volume (P<0.0001) and weight (P=0.0003). In U87 cells cultured in vitro, EPO treatment obviously accelerated the cell proliferation (P=0.020 on day 3 and 0.028 on day 5), promoted clone formation (P=0.0010), and increased proliferation index (P=0.0028); EPO significantly enhanced the protein expression of p-Akt (P=0.0020) and cyclin D1 (P=0.0022). The application of Akt inhibitor significantly suppressed the effect of EPO in enhancing cyclin D1 and p-Akt expression (both P<0.0001) and promoting cell proliferation.

CONCLUSIONEPO can significantly accelerate the proliferation of glioma through Akt pathway.

OBJECTIVETo adopt a criterion summarized in long-term practice to judge the immune status of recurrent aphthous ulceration (RAU) patients, and to analyze its rationality and feasibility.

METHODS60 cases were included, whose clinical information, such as age, sex, constitution status, history of system diseases and severity of ulceration were collected. The subjective judgement results of the immune status were obtained according to the criterion summarized in clinical practice. At the same time, peripheral blood parameters about immune function were examined in order to obtain the objective examination results of the immune status. With the examination result as the gold standard, the rationality of the criterion was estimated through its sensitivity and specificity.

RESULTSAccording to subjective judgment, there were 86.67% cases with abnormal immune status (28.33% cases with elevated immune status, and 58.33% cases with reduced immune status). According to objective examination, there were 93.33% cases with abnormal immune status (23.33% cases with elevated immune status, and 70.00% cases with reduced immune status. Sensitivity and specificity were 89.29% and 50.00% when the criterion was used to judge immune abnormality, 42.86% and 76.09% when the criterion was used to judge immune elevation, and 64.29% and 55.56% when the criterion was used to judge immune reduction.

CONCLUSIONIt is of some rationality and feasibility to judge the immune status according to the criterion based on clinical information, and the criterion can be used in small clinics.

Adult ; Female ; Humans ; Immune System ; physiology ; Male ; Stomatitis, Aphthous

Acute Disease ; Animals ; Ginkgo biloba ; Glutathione ; analysis ; Lung ; drug effects ; metabolism ; pathology ; Malondialdehyde ; analysis ; Paraquat ; toxicity ; Phytotherapy ; Plant Extracts ; therapeutic use ; Poisoning ; drug therapy ; Rats ; Superoxide Dismutase ; analysis ; Treatment Outcome

AIMTo study cyclosporin A (CsA) aerosol for anti-airway hyperresponsiveness (AHR) in sensitized rats.

METHODSSensitized Sprague-Dawley rats were given cyclosporin A (5, 20 g.L-1) and sodium cromoglycate (SCG, 20 g.L-1) by aerosol (5 min per day), dexamethasone (DXM, 0.5 mg.kg-1) i.p. once per day for 7 d before antigen challenge. The respiratory resistance(R(aw)) and lung dynamic compliance(Cdyn) of the rats induced by methacholine (Mch) were measured 24 h after ovalbumin(OA) challenge. The PC50 changes of R(aw) and PC25 changes of Cdyn were also investigated.

RESULTSPretreatment with CsA, sodium cromoglycate and dexamethasone inhibited the increase of R(aw) and decrease of Cdyn caused by inhaling Mch. The value of R(aw) PC50 in the CsA(5 g.L-1) group 5.6 g.L-1, the CsA(20 g.L-1) group 6.4 g.L-1, the SCG group 8.3 g.L-1 and the DXM group 9.2 g.L-1, was significantly higher than that of the model group 1.9 g.L-1 (P < 0.05). The value of Cdyn PC25 in the CsA(5 g.L-1) group 4.3 g.L-1, the CsA(20 g.L-1) group 5.4 g.L-1, the SCG group 6.4 g.L-1 and the DXM group 6.2 g.L-1, was significantly higher than that of the model group 1.1 g.L-1 (P < 0.01).

CONCLUSIONAnti-AHR of CsA by aerosol in animal model offered an experimental evidence for topical inhalation of CsA in treatment of asthma.

Administration, Inhalation ; Aerosols ; Airway Resistance ; drug effects ; Animals ; Cyclosporine ; administration & dosage ; pharmacology ; therapeutic use ; Disease Models, Animal ; Female ; Immunosuppressive Agents ; administration & dosage ; pharmacology ; therapeutic use ; Lung Compliance ; drug effects ; Male ; Ovalbumin ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiratory Hypersensitivity ; chemically induced ; drug therapy

Objective Mutated inbred animal model is introduced to the practical course of genetic diagnosis in the hope that medical students are able to apply what they have learned to clinical cases, based on a deep understanding of principle and technology on gene mutation detection. Methods We integrated DNA extraction, polymerase chain reaction, agarose gel electrophoresis, and gel imaging analysis into a comprehensive experiment and arranged 4-year-programme undergraduates majoring in preclinical medical sciences to conduct it with the purpose of investigating the internal relations between phenotype and genotype in a hairless Uncv mouse model. Subsequently, the questionnaire aimed at evaluating learning effect on the part of students was handed out and their feedbacks were analyzed. Results More than 90% of respondents are satisfied with the general learning effect. Especially, 98. 7% of students support the enhancing effect of the new teaching mode on their research skills and 96% consider the practical course helpful to their problem-solving ability. Conclusions The introduction of mutated inbred animal model to the practical system of molecular diagnostics proves beneficial to boost students' learning effect and scientific research quality. Our practice also provokes thoughts on the further utilization of animal models in teaching system of medical sciences.