Objective To study the effects of knock down midkine(MK)by siRNA on chemosinsitivity in bladder cancer cells. Methods Three MK siRNAs were designed and constructed. After transfected with MK siRNA or scrambled siRNA for different time, cultured cells were harvested to carry on the next experiments. Expression of MK was determined by real-time RT-PCR and western blot, and apoptosis were evaluated by caspase-3 activity and TUNEL assay. MTT was performed to examine the inhibition effect of Paclitaxel (PTX) on cells. Results MK siRNA could down-regulate the MK expression in a dose- and time-dependent manner. The MTT results showed that the inhibit rates were (18.21±0.36)%, (18.19±0.29)%, (17.89±0.33)%, (1.86±0.52)%, (32.56±0.53) %, (53. 83±0.38) % and (78. 95±0.55) % in different groups PTX alone(0.2μmol/L), ConA +PTX(0.2 μmol/L), Con-B +PTX(0.2 μmol/L), siRNA alone(12. 50 nmol/L), siRNA(3. 125nmol/L) + PTX (0. 2 μmol/L), siRNA (6. 25 nmol/L) + PTX (0. 2 μmol/L) and siRNA (12. 50nmol/L)+PTX(0.2 μmol/L), respectively. The TUNEL results showed that apoptosis index was (1.81 ±0. 36)%, (1. 89±0. 38)%, (5. 56±0. 58)%, (9. 68±0.55)% and (15. 25±0.56)% in different groups (Con-A, Con-B, siRNA (3. 125 nmol/L), siRNA (6. 25 nmol/L) and siRNA ( 12. 5nmol/L), respectively. The activity of caspase-3 increased significantly in transefected cells with a dose-and time-dependent manner. Conclusion MK siRNA could sensitize human bladder cancer cells to chemotherapy which might be through the apotosis.

Objective To construct shRNA against optineurin ( OPTN ) clone and transfect the construction into HEK293FT cells for inhibiting the expression of OPTN, which is laid the foundation for further research of molecu-lar mechanism OPTN protein. Methods The insert of this clone was a double-strained DNA sequence against OPTN which would be transcripted into shRNA and it was synthesized according to the sequence in Origene. Ex-pression vector pRFP-C-RS was employed to fuse the insert to get the construction and finally confirmed by sequen-cing. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN and RFP signals could be detected using fluorescence microscope. Western blot was further employed to check the protein level of OPTN. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN was then obtained to investigate the functional study of OPTN which was measured by Salmonella infection experiment. Results shRNA eukaryotic expression vector targeting OPTN gene was successfully constructed. HEK293FT cells were transfected with pRFP-C-RS-shOPTN and 72 hours later we observed strong RFP signals showing that shOPTN expressed. Western blot was further employed to check the protein level of OPTN which showed that OPTN expression was indeed interfered. Salmonella infection assays was then performed and showed that OPTN could inhibit the proliferation of Salmonella that has invaded HEK293FT cells. Conclusion HEK293FT cell line which expresses shRNA against OPTN show a sharp inhibition of OPTN protein expression by 80%. So this cell line can be further used to investigate how OPTN regulates auto-phagy as an autophagy receptor. Meanwhile we find that OPTN can be as a autophagy regulator and strongly sup-press the proliferation of invasive Salmonella in vivo.

Objective Alstr(o)m syndrome (AS) is a rare,autosomal recessive inherited disease characterized by various clinical manifestations.The aim of this study was to review the clinical characteristics and laboratory findings of AS.Methods Two cases of AS was reported.Combined with the clinical data of 7 cases of AS which had been reported in China,the clinical characteristics and laboratory findings of AS were reviewed.Results Visual disorder( median onset age:6.0 years ) and dysaudia( median onset age:10.3 years ) were found in 9 patients,short stature and obesity in 8 patients,acanthosis nigricans in 7 patients,diabetes mellitus( median onset age:14.5 years) in 6 patients,and heart disease in 4 patients; hyperuricemia was detected in 6 patients,hepatic dysfunction and hypertriglyceridemia in 5 patients.Conclusions Visual disorder was the first presentation in patients with AS.Deafness,obesity,diabetes,and short stature were common.These findings were helpful in making an early and accurate diagnosis and appropriate treatment.

Objective:To observe the effect of electroacupuncture at Baihui and Dazhui on cognitive function of vascular dementia rats and explore its possible mechanism. Methods:Male Sprague-Dawley rats were randomly divided into the sham operation group, model group and electroacupuncture group, with 10 rats in each group. Rats in the model group and the electroacupuncture group were made as vascular dementia models by carotid artery ligation of the two sides. After 2 weeks of modeling, the electroacupuncture group received electroacupuncture at Baihui and Dazhui for 4 weeks. Before and after the intervention, take the novel object recognition behavior test to detect the 1, 24 h new object preference coefficient; and use magnetic resonance spectroscopy to scan the levels of NAA/Cr and Cho/Cr neurometabolites in the hippocampus. q-PCR was used to detect the expression of caspase-3 and Bax in apoptotic cells. Immunohistochemistry was used to detect synapsin SYN expression. Results:Compared with the model group, the preference coefficient of new object of the electroacupuncture group at 1 h [(69.85 ± 11.26)% vs. (50.26 ± 8.85)%] and 24 h [(60.85 ± 11.86)% vs. (43.26 ± 10.25)%] significantly decreased ( P<0.01). The content of NAA/Cr (1.344 ± 0.149 vs. 0.892 ± 0.178) significantly increased ( P<0.01), and the content of Cho/Cr (0.661 ± 0.142 vs. 0.902 ± 0.178) in the hippocampus significantly decreased ( P<0.01). The expression of hippocampus caspase-3 (1.342 ± 0.041 vs. 2.437 ± 0.089), Bax (2.135 ± 0.083 vs. 4.358 ± 0.137) significantly decreased ( P<0.01), and the expression of SYN (0.577 ± 0.035 vs. 0.315 ± 0.025) significantly increased ( P<0.01). Conclusion:Electroacupuncture at Baihui and Dazhui can effectively improve the cognitive dysfunction of rats with vascular dementia, which may be related to the improvement of neuronal metabolites NAA, Cho content and inhibition of neuronal apoptosis and improvement of neuronal touch function.

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 investigate the relationship between preoperative high-sensitivity C-reactive protein (Hs-CRP) levels and clinical outcomes of patients undergoing off-pump coronary artery bypass graft (OPCABG) surgery.Methods:We prospectively selected 123 patients who received OPCABG at Beijing Anzhen Hospital from January 2019 to October 2019, and collected relevant preoperative and postoperative data. Patients were divided into a normal Hs-CRP group(78 cases) and an elevated Hs-CRP group(45 cases)according to the cutoff value (2 mg/L) of Hs-CRP level. The data of the two groups were compared, and regression analysis was performed on the postoperative data with differences to define independent factors.Results:The leukocyte count in the Hs-CRP group was significantly higher than that in the normal Hs-CRP group[(6.5±1.6)×10 9/ml vs. (7.4±2.1) ×10 9/ml, t=-2.839, P=0.005]. In the elevated Hs-CRP group, proportion of patients with atrial post-CABG atrial fibrillation (38% vs. 19%, χ2=5.100, P=0.024), duration of hospitalization[(21.2±7.1)days vs.(16.0±4.6)days, t=-4.469, P=0.000], hospital costs[(143.1±30.7)×10 3 yuan vs. (123.7±21.8)×10 3 yuan, t=-4.090, P=0.000]were significantly higher than those in the normal Hs-CRP level group. Smoking ( OR=1.660, 95% CI: 1.186-1.993, P=0.031) and Hs-CRP ( OR=1.170, 95% CI: 1.050-1.294, P=0.007) were independent risk factors for post-CABG atrial fibrillation. Hs-CRP ( B=0.436, 95% CI: 0.197-0.675, P=0.000) and left ventricular ejection fraction (LVEF, B=-0.180, 95% CI: -0.289--0.071, P=0.001) were independent influencing factors of duration of hospitalization. Hypertension ( B=-11.256, 95% CI: -20.670--1.842, P=0.020), Hs-CRP( B=1.235, 95% CI: 0.217-2.254, P=0.018) and LVEF ( B=-1.168, 95% CI: -1.634--0.702, P=0.000) were independent influencing factors of hospital costs. Conclusion:The preoperative Hs-CRP level of OPCABG is an independent influencing factor of post-CABG atrial fibrillation, duration of hospitalization and hospital costs. This finding lays the foundation for Hs-CRP combined with other indicators to accurately predict the prognosis of OPCABG and screen high-risk patients.

Objective To compare the prevalence and correlation factors of chronic kidney disease (CKD) in urban and rural areas in Minhang district of Shanghai through the social economic and clinical data of the elderly population.Methods Jiangchuan Street and Pujiang town were randomly selected to represent the urban and rural population in Minhang district of Shanghai,respectively.Based on the over-60-year old people health examination program,6151 objectives with complete clinical-epidemiological data and bio-chemical index were investigated.The prevalence of CKD in urban and rural areas was compared,and the correlation factors for the urban and rural CKD were evaluated by multiple logistic regression analysis.Results (1) The survey objectives with an average age of (69.57+7.04) years,including 4345 cases of the city residents and 1806 cases of rural residents,were enrolled.The age structures of urban and rural showed differences,population over 80 years old account for 13.1％ of the rural total,significantly higher than 7.4％ in the urban population (P ＜ 0.001).(2) The prevalence rates of diabetes,hyperuricemia,hyperlipidemia and hyperlipidemia in urban residents were higher than those in rural residents,which were 26.4％ vs 13.7％,9.9％ vs 2.3％,53.7％ vs 37.4％,51.4％ vs 15.6％ (all P＜ 0.01).The awareness rates of kidney disease and hyperlipidemia showed significant differences in urban and rural areas,which were 32.9％ vs 44.2％,84.6％ vs 62.8％ (all P ＜ 0.01).Compared with those in rural areas,the treatment rates of hypertension and high blood lipids in urban residents were increased (all P ＜ 0.01).(3) The prevalence of CKD was 23.4％.Female CKD prevalence was higher than male,respectively 26.3％ and 18.5％ (P ＜ 0.01).In urban CKD prevalence was 22.2％,lower than 25.2％ in rural.The prevalence rate of hematuria in urban areas was lower than in rural areas,but the prevalence rate of decline in renal function was higher (all P ＜ 0.05).With the increase of age,the prevalence rate of CKD was increased (P ＜ 0.01).(4) Age (OR=1.072),smoking history (OR=1.543),previous history of kidney disease (OR=1.351),diabetes (OR=1.373),hyperuricemia (OR=2.498),obesity (OR=1.364),history of interventional therapy (OR=1.896) had positive correlation with CKD in city elderly population,while the higher education (OR=0.676,OR=0.604) and drinking (OR=0.585) had negative correlation (all P ＜ 0.05).Age (OR=1.032),female (OR=1.860) had positive correlation with CKD in rural elderly population (all P ＜ 0.05).Conclusions CKD has been a common chronic progressive disease of the aged in Minhang district.The prevalence of CKD is higher in urban areas than in rural.Age is a common factor for CKD in urban and rural.Previous smoking,history of kidney disease,diabetes,hyperuricemia,obesity,history of interventional therapy,education and drinking have correlation with urban CKD patients.Female has correlation with rural CKD population.

Objective To study the effects of rehabilitation training on learning and memory ability and the expression of growth-associated protein-43(GAP-43)in rats with vascular dementia.Methods Forty-four female Sprague-Dawley rats were randomly assigned to a rehabilitation group(n=20),an immobilization group(n=20),and a sham-operation group(n=4).The rats in the former 2 groups were operated on to establish the experimental vascular dementia model by repeatedly ischemia/reperfusion injury of brain induced by ligation of bilateral common carotid arteries and lowering of blood pressure induced by intraabdominal injection of sodium nitroprusside.The rats in rehabilitation group were administered with rotating bar and rolling cage exercises for 1 hour once daily,while those in the immobilization group were immobilized and without any exercise:the rats in sham-operation group could move freely in cage.Learning and memory tests were preformed by using step-down avoidance test at the 27th and 28th days after operation.Immunohistochemical staining was used to detect GAP-43 expression in hippocampus CA1 area at different time points after operation. Results The rats in rehabilitation group demonstrated better learning and memory ability than those in immobilization group(P<0.01),and more GAP-43 expression in hippocampus CA1 ar-ea than those in immobilization group and sham-operation(P<0.01).Conclusion Rehabilitation training can im-prove the learning and memory ability of rats with experimental vascular dementia,and the mechanism is probably re-lated to the increase of GAP-43 in hippocampus CA1 area.

Objective To provide the endoscopic anatomic basis and anatomic parameters for endoscopic surgical therapy on orbital lesions , and to analyze the advantages and key points of this surgical approach .Methods Five fresh adult heads were used in this experiment .Endonasal thanslamina approach and endoscopic technique were applied to observe important anatomic marks while intraoperative medicalization of the medical rectus muscle was applied to observe the exposure and positioning of important structures and trends of the optic canal and intra orbit .Results Uncinate process was at the lower front corner of middle nasal concha; ethmoidalis bulla was behind the uncinate process , and ethmoidei sinus was reachable after an incision was made on ethmoidalis bulla;anterior ethmoidal artery and posterior ethmoidal artery were the important anatomic landmarks of the inner ethmoidei sinus;optic canal prominence , carotid artery prominence and OCR were the important anatomic landmarks of the inner sphenoid sinus; lamina papyracea was at the lateral wall of ethmoidei sinus , and orbital contents were approacchable after lamina papyracea was cut off ;inside orbit , the optic nerve was approachable through the gap between the medial rectus muscle and inferior rectus muscle .The ophthalmic artery of 9 out of 10 sides of the specimens was originated from the supraclinoidal segment of the internal carotid artery while the remaining one was originated from the cavernous segment of internal carotid artery .There were 7 sides in which ophthalmic artery was located at the inferior lateral of the optic nerve;there were 2 sides in which ophthalmic artery was located at the inferior of the optic nerve; the remaining one was located at the inferior medial of the optic nerve .Conclusion The endoscopic endonasal thanslamina approach can sufficiently expose the optic nerve and the structures in the medical space of the orbit.Uncinate process, ethmoid bulla, anterior ethmoidal artery, posterior ethmoidal artery and posterior ethmoid sinus are the important landmarks of the endoscopic endonasal thanslamina approach .Optic canal prominence , internal carotid artery prominence and OCR are the important landmarks for optic canal decompression .Ophthalmic artery , orbital branches , anterior ethmoidal artery , posterior ethmoidal artery , internal carotid artery are the important vessels . Medialization of the medial rectus muscle is effective to approach the orbital anatomical structures .

Objective To establish a close clinical and easy to operate animal artery bypass grafting model by using vascular anastomosis wheel.Methods 15 rabbits that weighted 2.5-3.5 kg were studied.Each animal underwent an end-toend anastomosis of jugular vein and carotid artery by vascular anastomosis wheel.Carotid ultrasound and flow detection were taken immediately and 2 months after surgery respectively,as well as morphology and pathology were recorded to analyze and evaluate the intimal hyperplasia of vein graft and arteriovenous anastomotic site.Results 14 rabbits were successfully established CABG model,however 1 rabbit died of respiratory inhibition caused of excessive anesthesia.Compared with normal carotid artery,the vein bridge showed significantly lower blood flow [(50.81 ± 1.33) ml/min vs.(70.59 ± 0.68) ml/min,P ＜0.01,higher PI(2.15 ±0.07vs.1.22 ±0.04,P ＜0.01)] immediately after surgery.Compared with the vein grafts immediately after surgery,the vein grafts 2 months after surgery showed significantly lower blood flow [(27.46 ± 2.15) ml/min vs.(50.81 ± 1.33) ml/min,P ＜ 0.01].Compared with normal jugular vein,the vein grafts 2 months after surgery showed significantly higher intimal hyperplasia[(160.30 ± 1.78) μm vs.(49.06 ± 2.76) μm,P ＜ 0.01],and higher number of elastic plates(12.36 ± 0.25vs.3.21 ± 0.15,P ＜ 0.01).Conclusion The use of vascular anastomosis wheel to establish an artery bypass graft model can imitate the pathological changes of vein grafts after CABG,which can provide an ideal animal model for various researches on vein grafts.