Objective To explore the preparation of specific immune RMA(iRNA) on anti-rabies and further study immunotherapy of rabies virus exposure. Methods Horses were immunized with the rabi-es virus and their livers were isolated from the horse of antiserum, from which total RNA was extracted and purified by sodium lauryisulfonate, phenol, chloroform, ethyiene glycol monomethyl ether, cetyltrimethyam-moniumbromide and alcohol. Results Pure preparation physico-chemical characterization was analyzed, and it's weight was 0.15% of weight of liver. The RNA contained 2.86% DNA and 1.16% protein. The iRNA with a maximum UV absorbance at 258 nm and A_(258/280) about 2.0. The test of RNA was positive, which had a relative molecular mas of 13.7×10~3 by high performance liquid chromatography(HPLC), and its hy-perchromic effect was 50.67%. The vesults of biological activity was showed that the rate of leucocyte adher-ence inhibition(LAI) was 41.73%, The protective rate was 50% and prolonging the life was 31.62%. Conclusion The results obtained with the practical value were identical and provide a basis on medicines of anti-rabies.

In order to obtain a more stable PEGylated interferon alpha-2b, and prolong its half life, interferon alpha-2b (IFN alpha-2b) was modified with monomethoxy polyethylene glycol propionaldehyde (mPEG-ALD) 20000. It was found that the optimized reaction condition for the maximum bioactivity and highest PEGylation degree of the mono PEGylated interferon alpha-2b was as follows: in 20 mmol/L, pH 6.5, citric acid and sodium dihydrogen phosphate buffer, the concentration of IFN alpha-2b was 4 mg/mL, and the molar ratio of PEG/IFN alpha-2b was 8:1, and the reaction time was 20 h at 4 degrees C. Under the optimized reaction condition, the mono PEGylation degree reached to 55%. Ion exchange chromatography was used to separate and purify mono PEGylated interferon alpha-2b from the reaction mixture. The purity of mono PEGylated interferon alpha-2b was higher than 97% characterized by HPLC. The bioactivity of the mono PEGylated interferon alpha-2b was 13.4% of the native IFN alpha-2b, while its half life in SD rat is much longer than the native IFN alpha-2b. The mono PEGylated interferon alpha-2b is also stable in aqueous.
Animals ; Antiviral Agents ; chemistry ; pharmacokinetics ; Drug Stability ; Humans ; Interferon-alpha ; chemistry ; pharmacokinetics ; Polyethylene Glycols ; chemistry ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins

Objective To evaluate the diagnostic value of circulating tumor cells(CTCs) in prostate cancer (Pca) through studying the relationship between CTCs and Gleason scores and pathological TNM stage in Pca patients. Methods A total of 238 patients including 161 Pca patients as cancer group, 35 male patients with benign prostatic diseases as benign group and 42 male with non-prostate disease as control group, who were treated in our hospital from July 2016 to January 2018,were enrolled. Venous blood of every patient was collected and CTCs were enriched and identified by immunocytochemistry CD45 capturing leukocyte and fluorescence in situ hybridization with chromosome 8 (CEP8-FISH). Cells displaying CD45-/DAPI+/CEP8>2 were characterized as CTCs. One-way ANOVA was used to exam the correlations of the number of CTCs with Gleason scores and pathological TNM stage. Results CTCs ≥2 were detected in 74.53％(120/161) of Pca patients and 20.00％(7/35)of benign prostatic diseases patients and 7.14％(3/42)of control group (χ2=79.605,P<0.05). In group Gleason scores 6, the numbers of CTCs were 2.00 ± 2.42, the ratios of CTCs≥5 and tetraploid were 13.33％ (2/15)and 26.67％(4/15) respectively. In 7 scores group, the results were 3.14±2.68,17.72％(14/79) and 34.18％(27/79)respectively;In 8 scores group, the results were 3.57 ± 2.70, 33.33％(7/21)and 42.86％ (9/21)respectively; In 9 scores group, these three results were 4.65±4.41, 43.48％(20/46) and 45.65％(21/46)respectively. The numbers of CTCs in the≤pT2b (20), pT2c(27), pT3a(19), pT3b(16)and≥pT4(12) groups were 2.25±2.45, 3.56±2.79, 4.05±3.47, 4.69±2.12 and 5.17±3.21 respectively. The ratios of CTCs≥5 were 25.00％(5/20), 25.93％(7/27), 26.32％(5/19), 50.00％(8/16) and 58.33％ (7/12)respectively. The proportions of tetraploid were 20.00％(4/20), 25.93％ (7/27), 31.58％(6/19), 50.00％(8/16) and 58.33％(7/12) respectively. There were significant differences between CTC and Gleason scores (F=3.200, P<0.05)and pathological stage (F=2.673, P<0.05). The ratios of CTCs≥5 increased with the increase of Gleason scores (χ2=11.592, P<0.05). Conclusions The detection of CTCs could be used for the differential diagnosis of Pca and benign prostatic disease. There were notable correlations between the numbers of CTCs and Gleason scores and pathological stage in Pca patients, especially between CTCs≥5 and Gleason scores.

Objective:To study the correlation between circulating tumor cells (CTC) and the degree of pathological invasion, recurrence and metastasis of urothelial carcinoma, and so to explore the clinical value of CTC detection in bladder cancer.Methods:A total of 142 patients with urothelial carcinoma in Huadong Hospital Affiliated to Fudan University were enrolled as cancer group from July 2016 to January 2018. According to the degree of tumor invasion, cancer group was divided into the non-muscle-invasive group (49 cases) and the muscle-invasive group(93 cases). In addition, 52 patients with benign urinary tract lesions admitted were selected as the benign group and 56 patients with non-urinary tract diseases and non-tumor as the control group. A total of 3.2 ml of venous anticoagulant blood from each subject was collected. CTC was enriched by negative enrichment using the magnetic beads coated with monoclonal antibody Cluster 45 of differentiation (CD45) to capture and remove white blood cells, and identified by chromosome 8 probe(CEP8) fluorescence in situ hybridization (FISH) technique. CD45-/4′,6′-diamidino-2-phenylindole+/CEP8>2(CD45-/DAPI+/CEP8>2) cells were judged as CTC. SPSS22.0 statistical software was used for statistical analysis.Results:≥2 CTCs/3.2 ml in blood was set as cutoff value. CTC positive rates in bladder cancer group, benign group and control group were 70.42%(100/142), 28.85%(15/52) and 8.93%(5/56), respectively, and there was a significant difference (χ 2=70.496, P=0.000). There was a statistically difference ( U=2 863.5, P=0.011) in the mean count of CTC(2 CTCs/3.2 ml vs 4 CTCs/3.2 ml) between the two groups. The proportion of≥5 CTCs/3.2 ml in the muscle-invasive group was 40.86% (38/93), which was significantly higher than that in the non-muscle-invasive group, 18.37% (9/49) (χ 2=7.330, P=0.007). Cystoscope follow-up of 65 patients treated with transurethral resection of the bladder tumor showed that the recurrence and metastasis rate in patients with≥5 CTCs/3.2 ml was as high as 47.62% (10/21), compared with 11.36% (5/44) of patients with<5 CTCs/3.2 ml (χ 2=10.530, P=0.001). Among 59 patients undergoing radical cystectomy, no significant difference was found in tumor diameter >3 cm, positive surgical margins and positive lymph nodes among all groups according to CTC negative or positive and CTC number ( P>0.05). But the recurrence and metastasis rate of patients with ≥5 CTCs/3.2 ml (59.10%) was significantly higher than that of patients with <5 CTCs/3.2 ml (6/30)(χ 2=8.364, P=0.004). Conclusion:The number of CTC increased with the deepening of tumor invasion; Tumor recurrence and metastasis increased significantly in the patients with ≥5/3.2 ml CTCs in blood.

Objective To explore the relationship between plasma microRNA 126 ( miR-126 ) level and coronary collateral circulation ( CCC) formation and to determine whether the miR-126 in plasma could serve as a blood-based biomarker for CCC in patients with severely narrowed coronary arteries ( CAD ).Methods In this prospective study , a total of 120 consecutive CAD patients with ≥95% stenosis in one epicardial coronary artery were enrolled.Thirty healthy people served as normal control.They were divided into two groups according to Rentrop grades:patients with grade 2 and 3 collateral development ( good CCC group, n=64) and patients with grade 0 and 1 collateral development (poor CCC group, n=56).Plasma miR-126 was measured by RT-PCR and serum VEGF was evaluated by ELISA method.Results Fasting plasma glucose ( FPG) was significantly lower in patients with good CCC than in patients with poor CCC ((5.99 ±1.48) mmol/L vs.(6.40 ±2.50) mmol/L).Plasma miR-126 levels and VEGF levels were significantly lower in CAD patients than in healthy people (0.04 ±0.01 vs.0.07 ±0.02, P=0.023 and (2 110 ±455) ng/L vs.(2 574 ±450) ng/L, P=0.011, respectively).miR-126 and VEGF levels were significantly higher in good CCC group than in poor CCC group ( miR-126:0.06 ±0.02 vs.0.03 ±0.01, P=0.021;VEGF:(2 549 ±614) ng/L vs.(1 759 ±452) ng/L, P=0.008).In CAD patients with good CCC, the miR-126 level was positively correlated to the VEGF expression (r =0.712,P=0.005) while there was no correlation between miR-126 level VEGF in CAD patients with poor CCC ( r =0.342, P =0.483).Multivariate analysis revealed that plasma miR-126 ( OR=2.145,95%CI 1.691 -2.988, P =0.001) and VEGF ( OR =1.279, 95%CI 1.068 -2.295, P =0.013 ) were independent predictors of collateral formation in patients with severely narrowed coronary arteries.In CAD patients , the area under the miR-126 ROC curve is 0.951 ( P=0.002).Conclusion Plasma miR-126 level is positively correlated to the CCC formation and is an independent predictor of CCC development in patients with severely narrowed coronary arteries , suggesting that plasma miR-126 might be a useful new , stable blood biomarker for predicting CCC formation in patients with severely narrowed coronary arteries.

Objective To observe whether there was difference of inferior glenohumeral joint capsule thickness(ICT)measured on coronal and transverse axillary section with high-frequency ultrasound.Methods ICT of 56 patients with frozen shoulder(FS group)and 115 healthy controls(HC group)were measured on coronal and transverse axillary sections with high-frequency ultrasound.The ultrasonic findings were compared between groups,while ICT measured on different sections were compared within groups.Results In FS group,ICT thickened,presented as low echo with poor boundary clarity,with reduced and uneven internal echo.No echo areas could be detected when there was fluid accumulation,and concomitant blood flow signal could be observed.In HC group,the inferior glenohumeral joint capsule presented as moderate echo with clear boundary,with uniform low or equal echo.No significant difference of ICT values measured on coronal or transverse section was found within both groups(both P＞0.05).Conclusion ICT measured on coronal and transverse axillary section with high-frequency ultrasound were not significantly different.

OBJECTIVETo explore the relationship between plasma microRNA126 (miR-126) level and coronary collateral circulation (CCC) formation and to determine whether the miR-126 in plasma could serve as a blood-based biomarker for CCC in patients with severely narrowed coronary arteries (CAD).

METHODSIn this prospective study, a total of 120 consecutive CAD patients with ≥ 95% stenosis in one epicardial coronary artery were enrolled. Thirty healthy people served as normal control. They were divided into two groups according to Rentrop grades: patients with grade 2 and 3 collateral development (good CCC group, n = 64) and patients with grade 0 and 1 collateral development (poor CCC group, n = 56). Plasma miR-126 was measured by RT-PCR and serum VEGF was evaluated by ELISA method.

RESULTSFasting plasma glucose (FPG) was significantly lower in patients with good CCC than in patients with poor CCC ((5.99 ± 1.48) mmol/L vs. (6.40 ± 2.50) mmol/L). Plasma miR-126 levels and VEGF levels were significantly lower in CAD patients than in healthy people (0.04 ± 0.01 vs. 0.07 ± 0.02, P = 0.023 and (2 110 ± 455) ng/L vs. (2 574 ± 450) ng/L, P = 0.011, respectively). miR-126 and VEGF levels were significantly higher in good CCC group than in poor CCC group (miR-126: 0.06 ± 0.02 vs. 0.03 ± 0.01, P = 0.021;VEGF:(2 549 ± 614) ng/L vs. (1 759 ± 452) ng/L, P = 0.008) . In CAD patients with good CCC, the miR-126 level was positively correlated to the VEGF expression (r = 0.712, P = 0.005) while there was no correlation between miR-126 level VEGF in CAD patients with poor CCC (r = 0.342, P = 0.483) . Multivariate analysis revealed that plasma miR-126 (OR = 2.145, 95% CI 1.691-2.988, P = 0.001) and VEGF (OR = 1.279, 95% CI 1.068-2.295, P = 0.013) were independent predictors of collateral formation in patients with severely narrowed coronary arteries. In CAD patients, the area under the miR-126 ROC curve is 0.951 (P = 0.002).

CONCLUSIONPlasma miR-126 level is positively correlated to the CCC formation and is an independent predictor of CCC development in patients with severely narrowed coronary arteries, suggesting that plasma miR-126 might be a useful new, stable blood biomarker for predicting CCC formation in patients with severely narrowed coronary arteries.

Biomarkers ; Collateral Circulation ; Coronary Angiography ; Coronary Artery Disease ; blood ; Coronary Circulation ; Coronary Disease ; Heart ; Humans ; MicroRNAs ; blood ; Multivariate Analysis ; Plasma ; Prospective Studies ; ROC Curve

OBJECTIVE To systematically evaluate the efficacy and safety of midazolam and dexmedetomidine/propofol for the sedation of critically ill patients undergoing mechanical ventilation， and to provide evidence-based reference for clinical treatment. METHODS Retrieved from PubMed， Embase， Web of Science， Cochrane Library， Clinical trials. gov， China Journal Full Text Database， Chinese Science and Technology Journal Database， Wanfang database and China Biomedical Literature Database， the data on the efficacy and safety of midazolam and dexmetomidine/propofol for the sedation of critically ill patients undergoing mechanical ventilation were collected from the establishment of the database to March 31， 2023. After extracting data from clinical studies that met the inclusion criteria， the meta-analysis was conducted by using the RevMan 5.3 statistical software. RESULTS A total of 31 literature were included， with a total of 2 765 patients. Results of meta-analysis showed that the mechanical ventilation time [MD＝14.13， 95%CI （13.75， 14.52）， P＜0.000 01] and the length of hospitalization in the intensive care unit [MD＝0.92， 95%CI （0.54， 1.30）， P＜0.000 01] of patients in the midazolam group was longer than dexmedetomidine/ propofol group. The incidence of bradycardia in midazolam group was lower dexmedetomidine/propofol group [OR＝0.60， 95%CI （0.41， 0.90）， P＝0.01]， but there was no statistically significant difference in the incidence of hypotension between the two groups [OR＝0.69， 95%CI （0.47， 1.01）， P＝0.06]. The incidence of delirium [OR＝3.88， 95%CI （2.74， 5.49）， P＜0.000 01]， ventilator- associated pneumonia [OR＝2.32， 95%CI （1.19， 4.51）， P＝0.01]， and respiratory depression [OR＝5.70， 95%CI （3.09， 10.52）， P＜0.000 01] in midazolam group were higher than dexmedetomidine/propofol group. CONCLUSIONS Compared with dexmedetomidine/propofol， midazolam increases patients’ mechanical ventilation time and the length of hospitalization in the intensive care unit in terms of efficacy， and increases the risk of delirium and pulmonary complications in terms of safety， but has a smaller cardiovascular impact.