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 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 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 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 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 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 evaluate the clinical efficacy of infliximab (IFX) in Crohn′s disease (CD) and its effects on mucosal healing and promoting fistula closure.Methods Between September 2007 and February 2011,relevant clinical data of CD patients treated with IFX in the Department of Gastroenterology,Ruijin Hospital were collected and the efficacy and safety of IFX were retrospectively analyzed.After IFX therapy,the efficacy evaluation included laboratory index,clinical efficacy,efficacy of fistula closure and mucosal healing.The data were analyzed using t test and Wilcoxon signed-rank test.Results A total of 22 patients were enrolled in this study,11 males and 11 females; the mean age was 29.3 years.The dosage of IFX was 5 mg/kg to 10 mg/kg at week 0,2,6to induce remission,and every 8 weeks on maintenance therapy.Of 22 patients,16 patients were active CD.One case dropped out.At week 14,of the remaining 15 cases,11 cases achieved clinical remission,two cases achieved clinically effective and two cases were ineffective.Crohn′s disease activity index (CDAI) (112±80) and ESR [(13±11) mm/1 h] of week 14 decreased compared with that of week 0 [(186±88),(21± 15) mm/1 h,P=0.04 and 0.007].Two cases of 10 patients with fistula dropped out as a result of ineffective,while eight cases had a partial response and six patients sustained response during the maintenance therapy,but no fistula closed and completely disappear.Seven patients reviewed by endoscopy after five times IFX therapy (24 weeks),after therapy the simple endoscopic score for Crohn′ s disease (SES-CD) ( 3.21 ± 2.89 ) decreased compared with that before treatment (5.86±3.02) (Z＝-2.38,P＝0.018).Eleven times of adverse events were found in nine patients,infusion reaction and respiratory tract infection were more common and no severe adverse effect was observed.Conclusions IFX can rapidly improve clinical symptoms and with good safety.The effects in mucosal healing and fistula closure may occur at early medication.

To evaluate the effects of San'ao decoction (SAD) and its analogous prescriptions (APs), compounds of traditional Chinese herbal medicine for asthma, on airway inflammation in mice with respiratory syncytial virus (RSV)- and ovalbumin (OVA)-induced asthma.

OBJECTIVE: To investigate the efficacy of CO2 laser treatment for patients with precancerous laryngeal lesions under phonomicrosurgery and to explore the points for attention in operation. METHOD: They were all treated with phonomicrosurgery techniques as mucosal epitheliumablation or mucosal stripping by using CO2 laser. Eight patients with laryngeal papilloma were excised by CO2 laser. RESULT: All patients were treated with CO2 laser surgery successfully. During follow-up of 6 to 39 months, all patients survived. Local recurrence or canceration were detected in 3 cases, of which 2 cases with laryngeal papilloma underwent CO2 laser treatment in one year post-operatively, while the other case with severe dysplasia underwent laryngeal vertical partial laryngectomy and post-operative radiotherapy one and half year postoperatively due to canceration. No local recurrence occurred until the last follow up. No severe complications such as dyspnea and hemorrhage occured. CONCLUSION CO2 laser surgery is an effective and minimally invasive treatment for precancerous laryngeal lesions. Through selecting the appropriate patient and paying attention to the operation during surgery, the adhesion of vocal cord can be reduced or even be avoided after CO2 laser surgery.
Adult ; Aged ; Carbon Dioxide ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Laryngeal Neoplasms ; surgery ; Laryngectomy ; methods ; Laser Therapy ; Lasers, Gas ; therapeutic use ; Male ; Microsurgery ; Middle Aged ; Precancerous Conditions ; surgery ; Treatment Outcome

As one of the commonest postoperative complications after liver transplantation,fungal infection has a high incidence and mortality.Nowadays,the development of the organ transplantation technique in China has already entered into the era of donation after citizen's death (DCD).Donors of DCD are mainly derived from the patients after brain death,cardiac death or death of both brain and cardiac.These donors usually suffer from long-term hypotension,ischemia,anoxia and secondary recessive or dominant infections during emergency treatment.Hence,preventing the fungal infections in donor organs and recipients after transplantation plays an important role in improving the success rate of liver transplantation.This review focuses on the clinical significance of preventing fungal infection in DCD of liver transplantation.