2.The efficiency and safety assessment of EntransterTM nanoparticle carrier for CD25 siRNA transfection in rat cornea
Qin, QIN ; Yunjie, SHI ; Min, ZHAO
Chinese Journal of Experimental Ophthalmology 2016;34(10):888-895
Background Gene transfection is an effective therapeutic avenue to target many kinds of eye diseases.Non-viral vectors with high transfection efficiency,long-term expression,low toxicity and high expression levels are pivotal in gene therapy of corneal disease.Objective This study was to evaluate and compare the safety and efficiency between EntransterTM and liposome vectors for transfer of CD25 siRNA in rat cornea.Methods Eighty male SPF SD rats were randomly divided into EntransterTM-CD25 siRNA group,liposome-CD25 siRNA group,simple CD25 siRNA group and normal saline solution (NSS) group with the right eye as experimental eyes.Corneal epithelia of the rats were completely removed after ocular surficial anesthesia,and 50 μl EntransterTM-CD25 siRNA,liposome-CD25 siRNA,CD25 siRNA solution and NSS were topical administered in the eyes respectively.Ocular response and green fluorescence number on the corneas were examined under the slit lamp assisted microscope 12 hours,24 hours,3 days and 7 days after use of the drugs.The rats were sacrificed and the corneas were obtained,and corneal histopathological examination was performed by using hematoxylin eosin stain.The gene transferred efficiency in the corneas was evaluated by fluorescence technology,and the safety of EntransterTM and liposome carriers was assessed using TUNEL stain.The expression and location of CD11b in the corneas were detected by immunofluorescence technology.The use and care of the experimental animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Committee.Results The quantity and intensity of fluorescence staining in the corneas were significantly increased in the EntransterTMCD25 siRNA group in comparison with the liposome-CD25 siRNA group,and the corneal fluorescence appeared earlier in the simple CD25 siRNA group,but it disappeared in 24 hours after transfection.Corneal histopathological examination revealed that the corneal edema and inflammatory cell infiltration in corneal epithelium after gene transfection were more dominant in the liposome-CD25 siRNA group than those in the EntransterTM-CD25 siRNA group,simple CD25 siRNA group and NSS group,and no abnormality was seen in the stroma and endothelium.The number of inflammatory cells was more in the liposome-CD25 siRNA group than that in the EntransterTM-CD25 siRNA group,simple CD25 siRNA group and NSS group (all at P =0.00).The number of apoptosis cells was significantly more in the liposome-CD25 siRNA group than that in the EntransterTM-CD25 siRNA group,simple CD25 siRNA group and NSS group in 12 hours and 3 days after transfection (all at P =0.00).Immunofluorescence assay showed the expression of CD11b primarily located in the corneal epithelial and stromal layers.The expression of CD11b was gradually enhanced over time in the liposome-CD25 siRNA group and peaked in 24 hours after transfection.However,the expression was absent in the EntransterTM-CD25 siRNA group,simple CD25 siRNA group and NSS group.Conclusions EntransterTM nanometer material-mediated transfection of CD25 siRNA in corneas of normal SD rats appears to have high transfection efficiency,low toxicity and slight irritating response to corneas,and EntransterTM vector is currently available for the gene therapy of corneal disease.
3.Satoyoshi's syndrome in a case.
Mei DONG ; Min WEI ; Shi-min ZHAO ; Hui-ping SHI ; Ding-jun NU ; Wei YU
Chinese Journal of Pediatrics 2004;42(3):224-224
4.The value of ultrasonography in diagnosis of hydropneumothorax
Ping SHI ; Yanni HAN ; Yan ZHANG ; Yan ZHAO ; Min PAN
Chinese Journal of Primary Medicine and Pharmacy 2009;16(4):594-595
Objective To investigate the value of ultrasound diagnosing for hydropneumothorax. Methods In a prospective double-blind randomized concurrent controlled trial. 213 patients doubted pneumotborax were exam-ined with CT, senography and conventional radiography. Results In 213 cases, hydropneumothorax diagnosed in 30 hemithoraces of 30 patients by CT,29 hemitboraces by ultrasound and 22 hemithoraces by X-ray. The sensitivity, nega-tive predictive value,accuracy by ultrasound and X-ray were 96.7% vs 73.3% ,99.8% vs 98.0% ,99.8 vs 98.1% respectively(P<0.05), the specificity and positive predictive value of both ultrasound and X-ray were 100%. Ultra-sound surpassed the X-ray in detecting pneumothorax ( McNemar test P<0.025 ). Conclusion If ultrasound is served to detect pneumothorax, it can make up the defects of the methods commonly used cuxrently.
5.Analysis of viral pathogen in children with acute lower respiratory tract infections
Man TIAN ; Shengyun SHI ; Min QIN ; Hongxia LIU ; Deyu ZHAO
Journal of Clinical Pediatrics 2010;(2):120-123
Objective To elucidate the etiology feature of viral infection in hospitalized children with acute lower respiratory infection. Methods A total of 5 480 children with acute lower respiratory tract infection, hospitalized from September 2007 to September 2009, were studied. Nasopharyngeal aspirates were screened for 8 types of viruses by direct immunofluorescence (DIF) assay. Results At least one type of viral pathogen was detected in 2 710 out of 5 480 patients and the overall positive rate was 49.5%. The most common virus was RSV (51.1%), followed by hMPV (18.9%), PIVⅢ (12.5%), ADV (7.1%), IFA (4.7%), IFB (2.9%), PIV Ⅰ (1.5%) and PIV Ⅱ (1.2%). The positive rate was highest in children under 6 months (43.5%). The seasonal change of RSV, hMPV was more obvious. The peak of RSV, hMPV appeared in the winter and the spring. The prevalence of viral infection in children with pneumonia, bronchitis, asthmatic bronchitis, non asthmatic bronchitis and asthma were 47.4%、63.6%、 50.5%、 30.1% and 43.5% respectively. Conclusions Viruses are the main cause of lower respiratory tract infections in children, especially in infants and young children. RSV and hMPV were the most common viruses in these years.
6.Ghrelin attenuates renal fibrosis and apoptosis of obstructive nephropathy
Guangxi SUN ; Lipei FAN ; Xianghua SHI ; Min LI ; Ming ZHAO
The Journal of Practical Medicine 2015;(7):1102-1106
Objective To investigate the effect and underlying mechanisms of ghrelin in a rat model of renal fibrosis. Methods Male Sprague-Dawley rats were divided into 4 groups , including sham operation +saline or brain gut peptide treatment group , model + saline or brain gut peptide treatment group. Unilateral ureteral obstruction (UUO) was established by left ureteral ligation. 7 days and 14 days after operation, the rats were sacrificed , while the kidney tissue of obstruction side was harvested for pathlogical changes through Masson coloration. Expression of α-smooth muscle actin (α-SMA), transforming growth factor beta1 (TGF-β1) and phosphorylated Smad3 (p-Smad3) in renal tissues were analyzed through immunohistochemistry. Expression of α-SMA and TGF-β1 mRNA was detected by real-time-PCR. Apoptosis kidneys cells were marked with TUNEL. Results Ghrelin inhibited renal fibrosis by reducing the production of collagen , restraining extracellular matrix (ECM) deposition and decreasing the expression of α-SMA. Meanwhile, ghrelin inhibited the accumulation of myofibroblasts by blocking the transforming growth factor-β1/Smad3 (TGF-β1/Smad3) signaling pathway. Moreover, ghrelin could attenuate renal tubular cell apoptosis induced by UUO injury. Conclusion Ghrelin can reduce renal fibrosis and renal cell apoptosis induced by UUO , demonstrating that ghrelin is a potent antifibrotic agent that may have therapeutic potential for patients with obstructive nephropathy.
7.Renal complications of hepatic glycogen storage disease
Min WEI ; Zhengqing QIU ; Hongmei SONG ; Shimin ZHAO ; Huiping SHI
Chinese Journal of Nephrology 2005;0(11):-
Objective To study renal involvement in hepatic glycogen storage disease(GSD) in childhood. Methods One hundred and eight patients aged less than 21 years old with type Ⅰa GSD (54 cases), type Ⅲ (29 cases) and uncertain type hepatic GSD (25 cases). Urine analysis, urine albumin, urine protein of 24 h, urine ?_2-MG, BUN, creatinine, Ccr were evaluated. Results Of 108 patients with hepatic GSD, 16 patients (20.8%) had proteinuria proven by urine albumin or urine protein of 24 h, their ages first found proteinuria were 8~15 years. Two 15-year-old patients had proteinuria over 1.0g/24h. Among 72 patients, urine ?_2-MG of 51 cases (70.8%) increased (175~10 623mg/L), and the mean urine ?_2-MG of type Ⅰ a GSD was much higher than that of type Ⅲ GSD, 4138.2 and 1790.1mg/L respectively. Of 91 patients, 10 had renal insufficiency, 1/10 (15-year-old girl) had heavy proteinuria (3.5g/24h), elevated BUN (9.3mmol/L) and Scr(1061 ?mol/L). Five elder patients (11~21 years old) had hematuria with renal colic caused by renal calculus. Conclusions Persistent protenuria, increased urine ?_2-MG, decreased Ccr, and renal stones are common complications of hepatic GSD in childhood. Renal function should be thoroughly evaluated during follow-up.
8.Expression of HBME-1,CK19 and CD117 in papillary thyroid carcinoma and its significance
Zheng ZENG ; Ruiping LI ; Min ZHAO ; Yanhua WEI ; Zhongyuan SHI
International Journal of Laboratory Medicine 2016;37(20):2825-2827,2830
Objective To explore the value of three antibodies in the differential diagnosis of papillary thyroid carcinoma ,by de‐tecting the expression of HBME‐1 ,CK19 and CD117 in papillary thyroid cacinoma ,thyroid follicular adenoma and Hashimoto′s thy‐roiditis tissues .Methods Totally 85 cases were collected from January 2013 to December 2015 ,including papillary thyroid cacino‐ma ,thyroid follicular adenoma and Hashimoto′s thyroiditis .They were immunohistochemical stained by HBME‐1 ,CK19 and CD117 .SPSS16 .0 software was used to analyze the relationship between the staining results with different pathological changes . Results The positive rates of HBME‐1 ,CK19 and CD117 were 87 .3% ,98 .2% ,and 7 .3% ,respectively .The positive expression of them in benign and malignant groups had significant difference (P< 0 .05) and their consistency checking Kappa were 0 .582 , 0 .551 ,and 0 .874 ,respectively .Conclusion In the differential diagnosis of papillary thyroid carcinoma and benign lesions ,CD117 is better than HBM E‐1 and CK19 .It′s possible to use a combination of them in practice .
9.Study on the Mechanism of Trimethoprim-Sulfamethoxazole Resistance in Stenotrophomonas Maltophilia
Suying ZHAO ; Hongxiang LIU ; Min LI ; Huiling CAO ; Hengfang SHI
Journal of Modern Laboratory Medicine 2014;(6):25-27,31
Objective Class 1 integrase(intI 1),qacE△1-sul1 and sul2 genes were detected in trimethoprim-sulfamethoxazole resistant Stenotrophomonas maltophilia by PCR to assess the relationship between the antibiotic resistance mechanism for trimethoprim-sulfamethoxazole (TMP-SMZ)and these genes distribution.Methods S.maltophilia isolates were collected from patients treated in Affilitated hospital of Nanjing University of TCM during January to May in 2013,DNA was ab-stracted by boiling method and genes were detected by polymerase chain reaction.Results intI 1 genes were observed posi-tive in 25 of 28 strains resistant for TMP-SMZ,qacE△1-sul1 genes were positive in 21 while sul2 genes positive in 15,the positive rates of intI 1,qacE△1-sul1 and sul2 genes were 89.29%,75% and 53.57%;in 18 trimethoprim-sulfamethoxazole sensitive strains,5 were intI 1 positive,4 were qacE△1-sul1 positive while sul2 were none,the positive rates of intI1 and qacE△1-sul1 were 27.78% and 22.22%.Conclusion Most of stenotrophomonas maltophilia resisted trimethoprim-sulfa-methoxazole had intI 1,qacEΔ1-sul 1 and sul2 genes.
10.Analysis of characteristics of ultrasound and fine needle aspiration cytology in patients with papillary thyroid carcinoma
Xiaojing ZHU ; Zhongyuan SHI ; Yanhua WEI ; Min ZHAO ; Jie LI
Tianjin Medical Journal 2017;45(5):前插3,533-535
Objective To analysis the diagnostic values and characteristics of ultrasound and ultrasound guided fine needle aspiration cytology (FNAC) in patients with papillary thyroid carcinoma (PTC) before operation. Methods The data of ultrasound and ultrasound guided FNAC in 129 patients (including 148 PTC nodules) with PTC were collected from January 2014 to February 2017, and the diagnostic reports and feature descriptions of ultrasound and ultrasound guided FNAC were analyzed retrospectively. Results Among the 148 PTC nodules, the ultrasonographic imaging showed that 84.5% (125/148) with low echo-level solid nodules, 61.5% (91/148) with echo heterogenicity, 77.7% (115/148) with a ratio ≥ 1 in longitudinal/breadth, 69.6 % (103/148) with fuzzy boundary, 75.0% (111/148) with microcalcification in nodules, 97.3% (144/148) without or with incomplete aureole, 64.9% (96/148) with rich blood flow and 7.0% (9/129) with enlargement of cervical lymph nodes. The FNAC diagnosis showed that 78.4% (116/148) was diagnosed with suspected papillary carcinoma, 1.4% (2/148) was diagnosed with malignent tumor, 11.5% (17/148) was diagnosed with atypia of undetermined significance (AUS), 1.4% (2/148) was diagnosed with benign lesion, 0.7% (1/148) was diagnosed with follicular neoplasm and 6.7% (10/148) could not be diagnosed. If the suspected papillary carcinoma and malignent tumor were defined as cytodiagnosis, the diagnostic accordance rate with intraoperative pathology was 79.7%. Conclusion The preoperative accuracy rates of ultrasound diagnosis and ultrasound guided FNAC diagnosis for patients with PTC are high, and the characteristics of the both are also typical. The two examinations before operation are helpful for early diagnosis and treatment formulation for patients with PTC.