Objective To investigate the effects and the possible mechanisms of berberine in human cervical cancer Hela cell line in vitro. Methods Cell growth rate was determined with MTT assay. TUNEL was used to examine cell apoptos rate. The expression of COX-2 and Bcl-2 proteins were detected by immunocytochemistry method. Results Berberine induced cytotoxicity and apoptosis in human cervical carcinoma Hela cells in time-dependent and dose-dependent manner. Berberine could down-regulated the Bcl-2 protein expression of Hela cells,but had no significant influence in the expression of COX-2 protein. Conclusion Berberine can inhibit Hela cell growth of cervical carcinoma and induce apoptosis in vitro. Down-regulation of the Bcl-2 protein expression may be involved in berberine-induced apoptosis of HeLa cells.

Objective To investigate the feasibility and promising applications of percutaneous transhepatic lymphosonography in detecting sentinel lymph nodes(SLNs).Methods Twenty five rabbits with VX2 tumor were included in this study.0.05 ml SonoVue was injected into the liver parenchyma at 12,3,6,9 points around the VX2 tumor.The situation of contrast-enhanced lymph-vessel emited from injected point and lymph nodes in hepatic portal or around tumor was observed,and then the position of the lymph nodes were detected with the help of the mark on the surface of the portal vein,caput pancreatis,collum vesicae biliaris.Methylene blue (MB) was injected in the same way as above.The injected points were massaged for five minutes,and then executed the experimental rabbits.The lymph nodes enhanced and all the lymph node dyed or not were taked out for recorded and pathologic examination.Results 34 SLNs were conformed by operation and pathological diagnosis in all the rabbits.All SLNs were confirmed pathologically,28 lymph nodes which were checked out by percutaneous transhepatic lymphosonography were all SLNs.In all the 31 lymph nodes which were checked out by MB,25 lymph nodes were SLNs and the rest were the second degree lymph nodes.The detection rate of percutaneous transhepatic lymphosonography (82.4％) and the MB (91.2％) showed no significant difference(P =0.169).There were 6 SLNs enhanced uniformitily in which 2 SLNs encroach by cancer cell and 22 enhanced asymmetrically in which 21 SLNs encroach by cancer cell.The sensitivity,specificity and accuracy of percutaneous transhepatic lymphosonography to detcect the SLNs benign or malignancy was 95.5％ (21/28),66.7％(4/6) and 89.3 ％ (25/28).Conclusions Percutaneous transhepatic lymphosonography is a reliable and noninvasive method to detect and estimate the SLNs of hepatic cancer.

Objective To investigate the role of mismatch repair gene hMLH1 in pancreatic carcinoma and its clinicopathological significance.Methods hMLH1 was extracted from 60 cases of pancreatic carcinoma tissues and 60 cases of normol pancreatic tissues.hMLH1 expression in pancreatic carcinoma and normal tissues was detected by SP immunohistochemical staining.Results The strong,weak and loss expression of hMLH1 in pancreatic carcinoma tissues and normal pancreatic tissues was 0 vs 83.33％ (50/60),31.7％ (19/60)vs 16.67％ (10/60),and 68.3％ (41/60) vs 0 respectively.The protein expression of hMLH1 was not related to patient's age,tumor location,or pathological types (P ＞ 0.05),but it was related to lymph node metastasis (x2 =8.579,P =0.004),clinical stage (x2 =9.586,P =0.002) and pathological differentiation (x2 =20.372,P =0.001).Conclusion The loss expression of hMLH1 has a correlation with pancreatic carcinogenesis,differentiation degree,and disease progression.

Objective To explore the teaching effect ofOrgan-system-based curriculum in-tegrated model on clinical graduation field work. Method 150 clinical medical undergraduates of Grade 2009 from Shenyang Medical College selected, were randomly divided into the experimental group (75 persons) and the control group (75 persons). The experimental group adopted the means of the Organ-system-based curriculum integrated model, namely practicing according to human organ-system and the control group accepted the traditional pattern of discipline centered practice during the clinical graduation internship. The two groups of students carried out questionnaire survey and comprehend examinations when the practice ended, and then statistics analysis (the chi square test, t test) was done by the SPSS 19.0 software between the two groups in order to explore the teaching effect. Result The students' satisfaction degree from six aspects of the questionnaire survey showed in the experimental group was significantly higher than that in the control group (the degree of recognition of practicing pattern: χ2=11.437, P=0.003; the architectonic integrality: χ2=9.881, P=0.007; the im-provement of the autonomic learning ability χ2=9.643, P=0.008; the teaching method and means: χ2=11.006, P=0.004; motivating learning interest: χ2=13.550, P=0.001; increasing the ability of clinical thinking: χ2=13.309, P=0.001), and the average test scores of students from three parts of examination results showed by comprehend examinations (speculative knowledge examination: t=2.768, P=0.006;technical skill examination: t=2.212, P=0.029; clinical capability examination: t=5.015, P=0.000) in the experimental group was obviously higher than that in the control group and the difference was sig-nificant. Conclusion Organ-system-based curriculum integrated model on clinical graduation in-ternship is generally approved by the students, which has significantly improved the students' clinical thinking ability, and the quality of graduation internship teaching.

ObjectiveTo observe the dynamic change of muscle tissue restoration after damaging at different observation time through acoustic radiation force impulse(ARFI).MethodsAccording to different observation time,16 healthy rabbits were randomly divided into four groups,including before injury,1 day after injury,7 day after injury,14 day after injury.A homemade gravity hammer was used to establish damage model of rabbit gastrocnemius,then applied ARFI to observe changes of virtual touch tissue quantification (VTQ) of gastrocnemius.All the rabbits were executed after measurement,then the pathological changes of muscle tissue were observed under microscope.Results The VTQ of damaged muscle group were significantly higher than that of nomal muscle group( P ＜0.01),and VTQ of 1 day,7day,14 day group after injuried had significant difference between two groups (P ＜ 0.01).Through pathological examination,normal musule fibers were continual and there were not swelling and bleeding.One days after injury,muscle fibers were fractured,swollen,bleeding and congestive.7 days after injury,large areas of muscle cells were necrotic,and amounts of calcium salt depositsed.14 days after injury,lots of fiber cells proliferated,and the deposits of calcium obviously reduced.The results of ultrasound elastography were consistent with these of pathological.ConclusionsARFI can direct,noninvasively evaluate the changes of muscle tissue restoration after different time of injury,and provide objective basis for diagnosis and treatment.

Objective To investigate the effect of ovulation induction of letrozole and clomiphene on the treatment of polycystic ovary syndrome.Methods From February 2014 to February 2014 80 cases with polycystic ovary syndrome in Hangzhou maternity hospitalas the research object were divided into two groups: the control group and the observation group.The control group were treated with clomiphene;the observation group were treated with letrozole.The clinical effect in the two groups were compared.Results The total effective rate was 95.0% in the observation group, which was higher than 67.5% in the control group, the difference was statistically significant(P<0.05).The thickness of endometrium in the observation group was(9.10±1.32)mm and(5.38±0.61)mm in the control group at the HCG injection day, the difference between the two groups was statistically significant (P<0.05).Conclusion Letrozole can be used in the treatment of polycystic ovary syndrome, can get high quality treatment effect, worthy of promotion.

Adult nursing course is the nursing professional core courses. The traditional spoon-feed teaching mode which lack of practice opportunities, can′t meet the social demand for“excellence” of the nursing personnel training. The teacher makes a combination of theory-practice-clinical consistent training mode and offline to online interactive learning mode, relying on high simulation training classrooms, teaching resources storehouse platform and mobile terminal real-time communication platform to construct the integrated teaching model of theory and practice. The course is based on clinical true cases, put the “task drive” and “problem solving” as the main line, pay attention to preview before the class, class learning, practice training and extension after class and other important time nodes. Students explored and practiced in three-dimensional learning environment to make up for the shortcomings of traditional teaching mode, and achieved good teaching results.

Objective The aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens. Methods Lung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical ( IHC) ALK( D5F3) detecting ALK protein expression was performed in 203 prepared
formalin?fixed paraffin?embedded ( FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid ( BL ) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT?PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization ( FISH) . Six patients with ALK IHC?positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments:( 1) Comparison of the results of 4% neutral buffered formalin fixed for different time ( 24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks;(2) Comparing qRT?PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens. Results Among the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results ( by at least one method), with an ALK test ratio of 90.4% (207/229).FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK ( D5F3) were successfully performed in all the 203 FFPE cell blocks ( 100%) , and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT?PCR, and 96 out of 98 ( 97. 96%) cytologic samples were successfully performed.18 out of 19 IHC ALK?positive cases were verified to be of ALK fusion status by qRT?PCR. The concordance rate was 94.7% ( Kappa=0.967, P<0.001) between Ventana IHC ALK( D5F3) and qRT?PCR, and the sensitivity of the Ventana IHC ALK ( D5F3) assay compared with qRT?PCR was 100%and the specificity was 98. 7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK(D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK(D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT?PCR test and ALK gene fusion showed good concordance. Conclusions The standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK( D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK( D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT?PCR may be an alternative option for the detection of ALK gene fusion.

Objective The aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens. Methods Lung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical ( IHC) ALK( D5F3) detecting ALK protein expression was performed in 203 prepared
formalin?fixed paraffin?embedded ( FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid ( BL ) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT?PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization ( FISH) . Six patients with ALK IHC?positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments:( 1) Comparison of the results of 4% neutral buffered formalin fixed for different time ( 24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks;(2) Comparing qRT?PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens. Results Among the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results ( by at least one method), with an ALK test ratio of 90.4% (207/229).FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK ( D5F3) were successfully performed in all the 203 FFPE cell blocks ( 100%) , and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT?PCR, and 96 out of 98 ( 97. 96%) cytologic samples were successfully performed.18 out of 19 IHC ALK?positive cases were verified to be of ALK fusion status by qRT?PCR. The concordance rate was 94.7% ( Kappa=0.967, P<0.001) between Ventana IHC ALK( D5F3) and qRT?PCR, and the sensitivity of the Ventana IHC ALK ( D5F3) assay compared with qRT?PCR was 100%and the specificity was 98. 7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK(D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK(D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT?PCR test and ALK gene fusion showed good concordance. Conclusions The standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK( D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK( D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT?PCR may be an alternative option for the detection of ALK gene fusion.

OBJECTIVE:To explore the association between the mitochondrial DNA(mtDNA)3537A/G,5351A/G variant and type 2diabetes mellitus(T2DM)in northem Chinese population.METHODS:The subjects including 614 patients with T2DM in Dalian City,61 of them were collected from family survey,497 of them were collected from Department of Endocrine,Dalian Municipal Central Hospital,and the remained 56 were selected from diverging T2DM patients in Dalian City.Additional 344 cases with normal carbohydrate toierance were served as controls.The mtDNA 3537A/G.5351A/G variants in 614 patients with T2DM and 334 healthy control subjects were examined.By sequencing the mtDNA in 24 cases and 26 controls,2 candidate SNPs in mtDNA were determined,and then genotyping was carried out by using PCR restriction fragment length polymorphism(PCR-RFLP)analysis.RESULTS:The frequency of mtDNA A3537G and A5351G mutation was 2.0%and 2.6% in T2DM patients,respectively,which was 2.1%and 4.2% in the control group.No significant difference had been observed between case and control(P＞0.05).After stratifying by body mass index and blood pressure,we found that the frequency of A5351 G in obesity patients with T2DM was 1.61%,and in obesity control was 15.38%,which had significant difference(P_(Fisher)=0.02,OR=2.76),however,A3537G stili showed no significant difference in all groups(P＞0.05).CONCLUSION:5351A/G in mtDNA ND2 gene may be a variance associated with T2DM in northem Chinese.