BACKGROUND: Multipotency of hepatic stem cells is of important value in liver transplantation. Stem cells have been successfully identified and isolated from the animal livers. However, reports on whether stem cells exist in human hepatic tissue and how to isolate and identify them ars few.OBJECTIVE: This study was in attempt to isolate hepatic stem cells from human para-cancerous tissues of hepatoma and in vitro culture them, also to identify the stem cell surface marker, in order to find a new source of heptatic stem cells.DESIGN: Cell observation experiment.SETTING: Department of Common Surgery, First Hospital, Jilin University; Department of Common Surgery,Dongfeng Hospital of Traditional Chinese Medicine.PARTICIPANTS: Samples were harvested from 10 patients with hepatoma admitted to Department of hepatobiliary surgery, First Clinical College, Jilin University between October 2005 and June 2006, with age of 45 to 58 years.Hepatic tissue 2 cm away from cancer nest was cut when patients underwent hepatectomy, and it was pathologically confirmed as carcinoma-free tissue. Written informed consents were obtained from each patient. DMEM/F12 dry powder used for cell culture was provided by Hyclone Company, USA. Fresh fetal bovine serum was prepared by Lianxing Biotech Co.,Ltd, Tianjin. Various cell growth factors were the products of Cytolay Company, USA.METHODS: Para-cancerous tissues of hepatoma was cut into pieces, rinsed with Hank's solution and digested with type Ⅳ collagenase. Then the isolated cells were re-suspended in the DMEM/F12 medium supplemented with 0.1 volume fraction of fetal bovine serum, and hepatocyte growth factors, epidermal growth factors and α- fibroblast growth factors of 25 μg/L each were added in the above medium. When the cultured cells covered 2/3 of bottom,they were digested with trypsinase for passage and inoculated at 2×107 L-1. When cells propagated to the 3rd and 4th generations, 2.60×109 L-1 cell suspension prepared with trypsinase was added, and subsequently, anti-human C-kit antibody, immunomagnetic beads and Buffer solution were added in order. C-kit+ cells were preliminarily isolated by immunomagnetic bead separation. Haematoxylin-eosin staining and immunofluorescent histochemical double-staining were used for detecting the hepatic stem cells in para-cancerous tissues.MAIN OUTCOME MEASURES: ① Observation of cell morphology. ② Identification of hepatic stem cells from para-cancerous tissues. ③ Identification of C-kit+ cells by immunofluorescent histochemical double-staining.RESULTS :① After primarily cultured for 2 weeks, the adherent cells grew in colony. After one half of culture medium was renewed, mature hepatocytes were gradually broken and disappeared. Small round cells propagated, and most of them were located in the center and arranged in cluster. Most cells were found with one big nucleus in each, less cytoplasm and clear cell boundary. When cells propagated to the 1st and 2nd generations, they still grew in colony, but fast. Each C-kit+ cell isolated by immunomagnetic bead separation presented a spherical cell body with a very big nucleus and less cytoplasm. After in vitro cultured for 1 week, it presented broken pieces and apoptotic symptoms.② After para-cancerous tissue was stained by haematoxylin-eosin, atypically proliferated biliary tracts with small round cells could be seen in the portal area. After para-cancerous tissue was stained by immunofluorescent histochemical double-staining, small round cells in the biliary tracts proliferated in the portal arsa co-expressed red fluorescence AFP and green fluorescence cytokeratin (CK) 19 with yellow superposition arsa. ③ After C-kit+ cells were stained by fluorescence immunocytochemisty, cytoplasm expressed alpha-fetoprotein (AFP) red granules and CK19 green granules. The superposition area of both presented yellow fluorescence of AFP+/CK19+-positive cells.CONCLUSION: Hepatic stem cells exist in human para-cancerous tissues of hepatoma. Therefore, expressions of C-kit+/AFP+/CK19+, the surface markers of hepatic stem cells, can be used for identifying and isolating hepatic stem cells. Small round cells obtained by in vitro isolation and culture, i.e. hepatic oval cells possess bipotential differentiation of hepatocyte and hepatobiliary epithelial cells.

Data were analyzed from self-filled questionnaires for the students who took the elective course of general practice in China Medical University. 95.5% of students thought it was necessary to study general practice in the college. 0-10 was used to score the degree of how they want to choose general practice as a career, the median was 0 before they had the class and the median was 5 after the class (Wilcoxon rank test, P<0.01). But there were still some places need to be improved in terms of curriculum design, teaching methods and community practice. The General Practice teaching need to be innovated and the teaching mode should be different from the traditional one.

Objective To investigate the relationships between Cyclooxygenase(COX-2) and angiogenesis and expressions of COX-2 and vascular endothelial growth factor(VEGF) in thyroid cancer and its clinical significance.Methods Immunohistochemistry was used to detect the expressions of COX-2 and VEGF in 60 thyroid cancer,(15 thyroid)adenomas and 10 normal thyroid tissues.Results The expression rates of COX-2 and VEGF in thyroid cancer were higher than those in thyroid adenomas and normal tissues(P

Objective To study local people's habits and customs concerning the prevention of dental fluorosis in an indoor coal-combustion-type fluorosis area of Zhaotong city Yunnan province, and to provide reference value for monitoring and control of the disease. Methods In 2008, 600 people of a 12-year-old group (400 people) and a 35 - 44-year-old group(200 people) were examined for dental fluorosis and oral health status in an indoor coal-combustion-type fluorosis area of Zhaotong. A questionnaire survey of 120 students and 60 adults was carried out on their habits and customs, stove changing status, knowledge of prevention of dental fluorosis, social psychology status, oral health behavior and their need for medical treatment. Results The total prevalence of dental fluorosis was 91.0% (546/600), dental fluorosis index was 2.58. The total prevalence of dental fluorosis of 12-year-old group was 86.5%(346/400), and dental fluorosis index was 2.12. The total prevalence of dental fluorosis of rural student was 90.5% (181/200), which was obviously higher in the rural group than the urban group [82.5% (165/200), χ2 = 5.48, P < 0.05]. The total prevalence of dental fluorosis of 35-44-year-old group was 100% (200/200), which was obviously higher than that of the 12-year-old group(χ2 = 29.67, P < 0.01). The rates of using coal, baking of food, changing stoves were 66.7% (60/90), 14.4% (13/90),57.8% (52/90), respectively, and these values of the urban were much lower than those of the rural[95.6%(86/90) ,31.1%(28/90) ,76.7%(69/90),χ2 = 24.51,7.11,7.29, all P < 0.01]. Only 23.3% (42/180) of people surveied had the knowledge of prevention of dental fluorosis, and the rate in the adult group[56.7% (34/60)]was higher than that of the student group [6.7% (8/120), χ2 = 55.9, P < 0.01]. The same rate in the urban student group was 0(0/60), and the rural student group was 13.3%(8/60), there was a significant difference between them(χ2 = 5.66, P < 0.05). Seventy-three point nine per cent( 133/180) of these people thought dental fluorosis had significant effect on their life, 91.1%(164/180) of these 'people wished to be treated. Eighty-four point four per cent( 152/180) of the people had a habit of toothbrushing,and 50.7% (77/152) of these people were still using toothpaste containing fluorine. Conclusions Zhaotong is a serious indoor coal-combustion-type fluorosis area. Enhancing health education to local people especially to students, and changing people's unhealthy habits and customs are keies to control the prevalence of indoor coal-combustion-type dental fluorosis.

Objective:To evaluate the effect of self-management treatment for patients with chronic Keshan disease in Gansu Province.Methods:From March to June 2018, 243 patients with chronic Keshan disease who were treated by self-management treatment outside the hospital in 7 Keshan disease counties in Gansu Province were selected as the survey subjects, and their general demographic and clinical data were collected. At the same time, the survey subjects were given a 6-month self-management treatment, and the self-management scale was used to evaluate the patients' self-management behavior before treatment and 6 months after treatment. The higher the score, the better the patients' self-management behavior. The influencing factors of self-management scale were analyzed by multiple linear regression analysis.Results:Excluding incomplete data, a total of 158 patients with chronic Keshan disease were included in this survey, including 96 males and 62 females. Compared with patients before self-management treatment, the patients' cardiac function classification and work ability improved significantly after 6 months of self-management treatment ( Z = - 4.685, - 5.934, P < 0.05); cardiothoracic ratio decreased (0.61 ± 0.08 vs 0.63 ± 0.09, t = 5.175, P < 0.05); in the cardiac color Doppler ultrasound indexes, left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were increased (0.41 ± 0.11 vs 0.36 ± 0.07, 0.21 ± 0.07 vs 0.18 ± 0.05, t = - 6.504, - 5.391, P < 0.05); the total score of the self-management scale was increased [(53.86 ± 9.29) scores vs (51.46 ± 10.50) scores, t = - 3.696, P < 0.05]. Further analysis of the efficacy of the patients' cardiac function classification showed that the patients' medication compliance was positively correlated with the efficacy of the cardiac function classification ( r = 0.243, P < 0.05). The result of multiple linear regression analysis showed that education level and efficacy of the cardiac function classification were influencing factors of the self-management scale for patients with chronic Keshan disease ( t = 2.466, 2.635, P < 0.05). Conclusion:The self-management treatment for patients with chronic Keshan disease can effectively improve heart function and improve self-management behavior.

Objective To explore the association of serum bilirubin level at the time of admission with the compos?ite outcome（disability or death）in discharged patients with acute ischemic stroke. Methods In a retrospective cohortstudy from June 1st 2009 to May 31st 2012, we continuously included 3151 patients with acute ischemic stroke and col?lected demography,lifestyle,clinical manifestations and laboratory test data. Functional outcome was measured with themodified Rankin scale (mRS) when subjects were discharged. Disability was defined as mRS≥3 and composite outcomewas defined as mRS≥3 or death. Serum bilirubin was divided into four groups according to the quartile. Multiple Coxregression analysis was used to assess the independent relation between serum bilirubin and disability death and the com?posite outcome. Results There were 407 disabled patients，the disability rate was 12.9%；and 104 patients were dead，the fatality rate was 3.3%.After adjusting for multiple factors, we found the risks of composite outcome with total bilirubin in the four quartile were higher than that in the first quartile, aHR and 95%CI were 1.335(1.047～1.702) respectively；The risks of composite outcome with indirect bilirubin in the four quartile were higher than that in the first quartile, aHR and 95%CI were 1.355(1.062～1.728) respectively; The risks of composite outcome with bilirubin direct in the third and the forth quartile were higher than that in the first quartile, aHR and 95% CI were11.403(1.089～1.807)and 1.431 (1.118～1.833) respectively.With the increase of total bilirubin，indirect bilirubin and direct bilirubin level，the compos?ite outcome of discharged patient was on the increase. Conclusions The study indicated that higher serum bilirubincould increase the risk of composite outcome in ischemic stroke patients, there was dose-response relationship ,and bili?rubin was a independent risk factor.

OBJECTIVE To provide laboratory evidence for the prevention and control of coagulase-negative staphylococci(CNS) and study the prevalence of meticillin resistance and vancomycin resistance in clinical CNS in our hospital.METHODS Meticillin-resistant coagulase-negative staphylococci(MRCNS) were detected with cefoxitin disk diffusion and mecA-PCR.CNS with reduced susceptibilities to vancomycin was detected with vancomycin agar screen test.Their MIC was determined with E-test and gene van was detected with multiplex PCR.RESULTS CNS was highly resistant to cephem,?-lactam,aminoglycoside,macrolide,and lincosamide.The confirm rate of cefoxitin disk diffusion to mecA-PCR was 96.2% and 92.3% of mecA in 52 CNS strains were detected.Four strains of staphylococci with heteroresistance to vancomycin(VRS) were screened in 256 CNS ones which were all MRCNS but without gene van detected.CONCLUSIONS It is important for clinical laboratory to detect VRS and MRCNS with suitable methods to prevent VRS infection and prevalence.

OBJECTIVETo study the correlation between serum insulin-like growth factor-1 (IGF-1) level and feeding difficulties in preterm infants.

METHODSA retrospective analysis was performed on 200 preterm infants born between January 2013 and January 2014. Venous blood samples were obtained within 24 hours after birth to determine the serum level of IGF-1. The correlation between IGF-1 level and feeding difficulties in preterm infants was analyzed by single-factor analysis and multivariate logistic regression analysis.

RESULTSThe serum IGF-1 level in the feeding difficulty group was significantly lower than that in the control group (28±4 ng/mL vs 63±8 ng/mL; P<0.05). Results of multivariate logistic regression analysis showed that high gestational age and birth weight were protective factors for feeding difficulties in preterm infants, whereas asphyxia, delayed initiation of feeding, use of aminophylline, perinatal infection and decreased IGF-1 level were risk factors.

CONCLUSIONSThe level of serum IGF-1 is correlated with feeding difficulties in premature infants. A reduced IGF-1 level increases the risk of feeding difficulties.

Birth Weight ; Breast Feeding ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; blood ; Insulin-Like Growth Factor I ; analysis ; Logistic Models ; Male ; Retrospective Studies

OBJECTIVE To define the regularity of survival ability of HIV in natural environment,and prevent(infection) through contacting with positive body fluids during daily life or medical work.METHODS Having been diluted by sterile water or 10% serum RPMI 1640 medium,HIV was exposed to 4℃,room temperature(20-26℃) or 37℃ for different period of time.TCID_(50) of these samples was detected.Non-pathological samples were blind passaged for three generations.RESULTS HIV infective ability persisted more than 35 days both in(water) and medium at 4℃;whereas it persisted 7-14 days in water,14-21 days in medium at room temperature and 37℃.CONCLUSIONS HIV has higher resistance in natural environment.To prevent accidental spreading of HIV,HIV positive liquids and contaminants staffs should be treated carefully.

Objective:This study investigates the significance of human papilloma virus (HPV) subtype detection in opportunis-tic screening for cervical cancer in Uygur and Han women. Methods:Flow-through hybridization gene chip and thin-prep cytology test were used to detect HPV in cervical cell samples from 1140 females. A total of 428 patients had undergone cervical biopsies through colposcopy. The diagnostic results of the HPV subtype test for cervical lesions were evaluated on the basis of histology. Results:Total HPV infection rate for the 1140 cervical samples was 30.3%. The most common HPV subtypes were HPV16, HPV58, HPV52, HPV18, and HPV45. HPV52 infection rate was higher in Han women than in Uygur women, with statistically significant differences between the two (χ2=8.737, P=0.003). Among these cervical samples, the single HPV infection rate was 22.4%(255/1140), whereas the multiple HPV infection rate was 6.1% (69/1140). The sensitivity and specificity of the HPV subtype test for cervical lesions were 86.4% and 24.5%, respectively. The positive and negative predictive values were 58.5%and 59.3%. Conclusion:HPV infection subtypes in Ugyur and Han women have unique characteristics. Subtype detection is important in opportunistic screening for cervical cancer.