Objective: To observe the expression of CD133 in the clear cell renal cell carcinoma (RCC) and to investigate the related drug resistance. Methods: Flow cytometry and immunohistochemical method were used to examine the expression of CD133 in metastatic RCC cell line RCC05-TXJ, low metastatic RCC cell line RCC05-ZYJ and two clinical non-metastatic RCC primary cultures isolated from a male and a female patient. The four cell lines were treated with IFN-α and 5-FU and the viability of cells were examined by MTT assay before and after treatment. Results: Flow cytometry showed that RCC05-TXJ and RCC05-ZYJ expressed CD133. The in situ carcinomas of male and female patients hardly expressed CD133. Immunohistochemistry examination showed that the local membrane of RCC05-TXJ and RCC05-ZYJ cells expressed CD133. RCC cells of female and male patients hardly expressed CD133. RCC05-TXJ and RCC05-ZYJ cells exposed to IFN-α or 5-FU showed a rebound of survival 24 h after withdrawal of drugs. The survival rates of RCC cells of male and female patients kept at a low level after withdrawal of drugs. Conclusion: There are a small number of CD133+ RCC cells in RCC patients, with a property similar to tumor stem cells, which might be one of the important factors influencing the immunotherapy and chemotherapy of tumors.

Objective: To establish clear cell renal cell carcinoma (ccRCC) cell lines from clinical ccRCC specimens of Han nationality in China and to characterize the biological features. Methods: From 2005 to 2007, fresh surgical samples of ccRCC were obtained from 43 patients; the samples included primary tumor in situ, osseous metastasis, lymph node metastasis, and cancerous embolus. The samples were cultured in vitro using explant-culture method within 30-60 min after surgery. Analysis on cell growth and colony-forming efficiency was recorded for the lines which were passaged for over 50 generations. Chromosome examination, pathological examination and tumorigenesis in NOD-SCID mice were used to determine their malignancy. Flow cytometry was used to determine expression of CA9 and CD133. Results: Most of the primary cells could only be passaged for less than 5 generations; 5 lines could be serially passaged for over 5 passages, 3 lines for over 10 passages, and only 2 lines could be stably passaged. One line,named RCC05-TXJ, was from osseous metastatic ccRCC and had been serially passaged for 110 generations in 21 months; the average doubling time was 19.2 h,average chromosome number was 75,and colony forming efficiency was 41%. Another line,named RCC05-ZYJ,was from primary ccRCC specimen and had been serially passaged for 160 generations in 18 months; the average doubling time was 16.5 h,average chromosome number was 55,and the colony forming efficiency was 37%. Immunohistological analysis demonstrated that both lines expressed CA9 and CD133. Flow cytometry analysis found that expression levels of CA9 and CD133 increased with the passages. Both RCC05-ZYJ and RCC05-TXJ lines were able to form tumor and to metastasize in NOD-SCID mice; however, their metastatic ability was obviously different. Conclusion: We have established 2 ccRCC cell lines with different metastatic potentials from the clinical ccRCC specimens of Han nationality in China. The ratio of tumor stem cells increases with the passages.

Adipocytes ; cytology ; Cell Line ; Endothelial Cells ; cytology ; drug effects ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Melatonin ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism

Objective To study the effect of granulocyte-macrophage colony-stimulating factor(GM-CSF)on angiogenesis of rat with acute myocardial injury induced by isoproterenol(Iso). Methods A total of 60 adult male Wistar rats were randomly divided into 3 groups: normal control group, GM-CSF pretreatment group (GM-CSF group), and lso injury group, 20 rats in each group. GM-CSF group was administered recombinant human(rh)GM-CSF(5.0 μg/kg), through tail intravenous injection once a day for three days. Then the GM-CSF group and the Iso injury group were anesthetized by intraperitoneal injection of lso( 15.0 mg/kg) once a day for three days. The same dose of saline was administered in the same way to the control rats. Ten days after injection, pathological changes of myocardial damage and infarct area were examined by immunohistochemistry. The mRNA expression levels of polypeptide antigen (CD34), vascular endothelial growth factor (VEGF) and its receptor KDR/flk- 1 were measured by RT-PCR. Results The difference of myocardial necrosis area between groups was statistically significant(F=10.07, P < 0.01), in which GM-CSF group[(37.37 ± 12.98)%] was significantly less than Iso injury group[(45.51 ±14.96)%, P < 0.05]. The difference of myocardial neovascularization density index of rats between groups was statistically significant ( F = 25.54, P < 0.05 ), in which GM-CSF group [(3980.05 ± 477.22) No/mm2] was significantly higher than Iso injury group((2605.93±361.49)No/mm2,P<0.01).The differences of myocardial CD34,VEGF,KDR/flk-1 mRNA expression between groups were statistically significant(F=17.83,4.29,4.10,all P<0.01).Compared to Iso mjury group[CD34(23.85±6.06),VEGF(31.80±8.05),KDR/flk-1(30.16±8.01)]were higher in the GM-CSF group[CD34(44.04±10.13),VEGF(49A±11.59),and KDR/flk-1(46A9±7.90),all P<0.01].The expressions of myocardiM VEGF mRNA and its receptor KDR/flk-1 mRNA was positively correlated(r=0.725,R2=0.526,P<0.01).Conclusions GM-CSF prelreatmcnt increases the density ofnew blood vessels in myocardium,and reduces the Iso-induced myocardial injury in rats.

Objective: To learn the epidemiological characteristics of infectious disease related public health emergencies in Zhejiang Province from 2010 to 2018 for the prevention and control. Methods: The surveillance data was extracted from National Public Health Emergency Management Information System. Descriptive epidemiology method was used to analyze main diseases as well as distribution characteristics of time and places. Results: A total of 445 events were reported, which caused 14362 cases and 34 deaths, with a attack rate of 0.69% and mortality rate of 0.24%. There were 298 events with less than 30 cases, accounting for 66.97%. The event classification was dominated by general events ( 242 events, 54.38% ) and ungraded events ( 201 events, 45.17% ). The main diseases were chickenpox ( 134 events, 30.11% ), hand foot mouth disease ( 59 events, 13.26% ) and other infectious diarrhea ( 51 events, 11.46% ). The incidence peaked from April to June ( 129 events, 28.99% ) and from November to December ( 131 events, 29.44% ). Ningbo ranked the top in the number of reported events ( 141 events, 31.69% ). Most events ( 322 events, 72.36% ) occurred in schools. Conclusions The infectious disease related public health emergencies in Zhejiang Province from 2010 to 2018 were mainly caused by chickenpox, hand-foot-mouth disease and other infectious diarrhea. The two peaks of the emergencies occurred from April to June and from November to December. Ningbo was the main area reporting infectious diseases, and schools were the main places.

Objective To study the effects of microRNA-34a-5p on erythroid differentiation of K562 cells.Methods K562 cells were transfected with the microRNA-34a-5p mimics and antisense inhibitors specifically targeting mi-croRNA-34a-5p, respectively.The effects of over-expression or knocking-down of microRNA-34a-5p were exam-ined by Quantitative RT-PCR.Flow cytometry was performed to detect specific surface marker of erythroid cells . The benzidine staining assay was used to access the differentiation of K 562 cells.Western blot was performed to de-tect miRNA targets.Results microRNA-34a-5p was down-regulated at the early stage of K562 erythroid differenti-ation.Over-expression of microRNA-34a-5p in K562 cells attenuates erythroid differentiation , in contrast, inhibi-tion of microRNA-34a-5p accelerates erythroid pheotypes in K562 cells.c-MYB was found to be the direct target of microRNA-34 a-5 p in erythroid cells .Conclusions microRNA-34 a-5 p regulates early erythroid differentiation of K562 cells via repressing c-MYB.

Kidney injury and decreased chemosensitivity of tumor cells are obstacles with cisplatin (CDDP) chemotherapy. Down-regulation of the organic cation transporter 2 (OCT2) and multidrug resistance-associated protein 2 (MRP2) is a key means to alleviate CDDP-induced kidney injury and increase chemosensitivity. Astragaloside IV (AS IV) is obtained from the well-known traditional Chinese herb Astragalus membranaceus. This study explored the role of AS IV in preventing kidney injury and enhancing the antitumor effect of CDDP by suppressing OCT2 expression in kidney and MRP2 in tumors. This project was reviewed and approved by the Animal Ethics Committee of the First Hospital of Jilin University. The effects of AS IV on CDDP inhibition of tumor growth and promotion of apoptosis were assessed in Lewis lung tumor (LLC)-bearing mice by H&E and TUNEL staining. Kidney injury was assessed by serum biochemical parameters and H&E staining. We used Western blotting and immunohistochemistry assays to detect OCT2 and MRP2 expression in kidney and tumor. The concentration of CDDP in kidney and tumor was measured by HPLC-MS/MS. AS IV enhanced CDDP chemosensitivity by increasing tumor cell apoptosis and slowing tumor growth, and decreased kidney injury as evidenced by lower blood creatinine (Cr) and blood urea nitrogen (BUN). Co-administration of AS IV suppressed MRP2 overexpression induced by CDDP in tumor tissues and may be an important mechanism for enhancing CDDP chemosensitivity. Moreover, AS IV reduced CDDP-induced kidney injury in mice along with suppression of OCT2 expression in kidney. The concentration of CDDP was increased in tumor but decreased in kidney. In total, AS IV not only enhanced the antitumor effect of CDDP by suppressing MRP2 expression in tumor cells, but also decreased kidney injury induced by CDDP. The results provide new insight into the combined use of a chemotherapy drug and natural ingredients to treat cancer.

OBJECTIVETo compare the clinical efficacy on chronic pelvic inflammation between the acupoint embedding therapy and acupuncture, and to compare the efficacy of different patterns/syndroms in differentiation treated with acupoint embedding therapy.

METHODSTwo hundred and eighteen cases were randomized into an embedding therapy group (115 cases) and an acupuncture group (103 cases). In both groups, Shenshu (BL 23), Guanyuanshu (BL 26), Zigong (EX-CA 1), Yaoyangguan (GV 3), Guanyuan (CV 4) and Qihai (CV 6) were selected as the main points. For qi and blood stagnation pattern/syndrome, Zhongdu (LR 6), Diji (SP 8) and the others were added; for cold and damp stagnation pattern/syndrome, Diji (SP 8) and Sanyinjiao (SP 6) were added; for stasis and phlegm pattern/syndrome due to spleen deficiency, Sanyinjiao (SP 6) and Zusanli (ST 36) were added. In the embedding therapy group, the catgut was embedded at 7 - 13 acupoints each time, once every 10 days. In the acupuncture group, the conventional acupuncture therapy was applied, once a day. The symptom scores were assessed in the aspects of the lower abdominal pain severity and attack frequency, lumbosacral soreness and distention, abnormality of vaginal discharge and the others. The efficacy was compared between the two groups.

RESULTSThe total effective rate was 93.0% (107/115) in the embedding therapy group, which was better than 83.5% (86/103) in the acupuncture group (P < 0.05). The symptom scores were all reduced after treatment in the two groups (all P < 0.05). In the embedding therapy group, the efficacies were not significantly different among different patterns/syndromes (all P > 0.05).

CONCLUSIONThe acupoint embedding therapy achieves the definite efficacy on chronic pelvic inflammation and obtains the similar efficacies among the different differentiated patterns/syndromes of the disease.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Catgut ; utilization ; Chronic Disease ; therapy ; Diagnosis, Differential ; Female ; Humans ; Middle Aged ; Pelvic Inflammatory Disease ; diagnosis ; therapy ; Prostheses and Implants ; Treatment Outcome ; Young Adult

Objective:To investigate the common cause of spinal surgery in ankylosing spondylitis (AS) and to develop reasonable and effective treatment programs for rhematologists.Methods:In this study,79 AS patients (72 males,7 females) hospitalized in the Department of Orthopedic and received spinal surgery in Peking University Third Hospital from January 2007 to July 2013 were investigated retrospectively.The causes of the spinal surgery were analyzed.Results:The majority of the surgery were of cervical vertebra surgery for 59 cases (74.7％),followed by the thoracolumbar vertebra,thoracic vertebra,cervicothoracic vertebra,and lumbar vertebra.The common causes of AS spinal surgery were instability (53.2％) including fracture and dislocation caused by trauma mostly,followed by compression symptoms (34.1％).However,the orthopaedic surgery (12.7％) was not the predominant surgery causes.The most common cause of cervical surgery was cervical spondylosis (25 cases),followed by cervical fracture (22 cases) and cervical dislocation (10 cases).For cervical fracture,the most common site was C7 (8 cases).Atlanto-axial vertebral lesions (13 cases) induced by dislocation,subluxation,instability and fracture were common in cervical vertebra surgery.The age and disease duration of atlantoaxial surgical patient's were less than other parts of the cervical surgery.Thoracolumbar vertebra surgery included thoracolumbar kyphosis orthopaedic surgery (10 cases),and fractures (6 cases),which mainly occurred in T11-12 vertebrae.According to the time of admission for surgery,the cases of AS that received spinal surgery were 34 from 2007 to 2008.The number of the cases that accepted the surgery decreased gradually year by year.Conclusion:The majority surgery section was cervical vertebra,and atlanto-axial vertebral lesions were not rare as we thought.The main etiology of spinal surgery for AS patients was instability,e.g.fracture and compression symptoms.Of course,trauma accounted for the relative percentage of this phetonomenon.Although,the annual number of patients who receives spinal surgery is decreasing,to go to make early diagnosis and treatment,and to standardize patient education for AS patients are still important.

Objective To explore the relationship between pulse blood pressure and cognitive function for the elders in Yuhuan County,Zhejiang Province.Methods Multi -stage cluster method was used to select 1 700 residents aged over 60 years in Yuhuan County,Zhejiang Province.Mini -Mental State Examination (MMSE)was used to screen cognitive impairment.Locally weighted scatter plot smoothing was used to explore the local linear trend between MMSE score and diastolic.The association between pulse pressure or blood pressure and cognitive function was analyzed by multivariate logistic regression analysis.Results The prevalence of cognitive impairments for the elders aged over 60 years in Yuhuan County was 15.74%.There was significant difference between the pulse blood pressure of cognitive impairment group (64.01 ±16.31)mmHg and none -cognitive impairment group (58.97 ±14.67)mmHg.The t value was 4.97.Pulse blood pressure and the elderly MMSE score showed a significant positive correlation.Multivariate logistic regression analysis showed that cognitive function related to diastolic blood pressure (OR =1.010,95% CI:1.001 -1.019,P =0.034)after adjusting several covariate.Conclusion Normal cognitive function was associated with decreased pulse blood pressure among the elders aged over 60 years in Yunhuan County,Zhejiang Province.