ObjectiveTo investigate the expression of miR-155 and miR-146a in peripheral blood mononuclear cells (PBMC) and plasma of rheumatoid arthritis (RA) patients.MethodsPBMC and plasma were separated from the peripheral blood of 34 RA patients and 15 healthy individuals.Total RNAs were isolated and miRNAs were purified.The levels of miR-155 and miR-146a were determined by quantitative reverse transcription PCR(qRT-PCR).U6 was used as housekeeping control.The amount of target miRNA was normalized relative to the amount of U6(ΔCt=ΔCtmiRNA-ΔCtU6).Relative expression levels were expressed as 2 △-ΔCt.Data were analyzed using SPSS 13.0 software.The test of homogeneity of variance and unpaired t-test was used to compare between groups.P values(2-tailed) less than 0.05 were considered as statistically significant.ResultsThe expressions of PBMC and plasma miR-155 were higher in RA patients than those in the healthy control individuals(0.08±0.08 vs 0.05±0.03,t=-2.225,P＜0.05; 5.9±6.7 vs 1.3±2.0,t=-3.677,P＜0.05).The expression of miR-146a in PBMC and plasma of RA patients and controls were(1.3±1.2 vs 0.8±0.6,t=-2.154,P＜0.05)and(741±1001 vs 300±295,t=-1.669,P＞0.05).According to their DAS28 value,RA patients were divided into high activity group (23 cases,DAS28≥5.0) and low disease activity group( 11cases,DAS28＜5.0).The plasma miR-155 and miR-146a expressions were significantly higher in high activity group than those in low activity group.There were no significant differences in the expression of PBMC miR-155 and miR-146a between the two groups.ConclusionThe expression of PBMC and plasma miR-155 and miR-146a are higher in RA patients.The expression of plasma miR-155 and miR-146a are associated with RA patients' activity.Plasma miR-155 and miR-146a may be potential non-invasive biomarkers for RA diagnosis anddisease activity assessment.

ObjectiveTo assess the recent effect of the invasiveness and transsphenoidal microsurgical operation in the treatment of growth hormone-secreting adenom tumor.MethodsThe data of 22 patients with growth hormone(GH) adenomas were collected.All patients were tested preoperative GH level and that within the first 3 days,30 days following surgery.The invasiveness of pituitary adenomas was judged according to Hardy's classification scheme modified by Wilson.ResultsThere were 22 patients with GH-secreting pituitary adenomas,8 males and 14 females,11 patients with invasive adenomas and 11 patients with noninvasive adenomas.The remission rate between the patients with invasive adenomas and the patients with non-invasive adenomas had no statistically significant difference (P ＞ 0.05 ).The postoperative remission rate of patients with GH values ≤ 50μg/L was no significantly lower than that of ＞ 50μg/L( P ＞ 0.05).ConclusionThe recent effects of operation were not significantly different with the invasiveness of tumor.

Objective To summary the experience of microsurgery of meningioma in skull base treatment. Methods To study the clinical data of 10 cases patients of meningioma in skull base who were subjected to microsurgery by using retrospective analysis. Among the 10 patients, there were 3 cases of meningioma of the olfactory groove. There were 2 cases of sphenoid ridge meningioma, meningioma in the middle cranial fossa, meningioma in the cerebellopontine angle respectively.There was 1 case of meningioma of velarium in temporal lobe. Results In 7 cases, the meningioma was completely removed. And in another 3 cases, the meningioma got subtotal resection. During and after the operation, there were no severe complications observed or death occurrence in all the 10 cases of meningioma patients. Two months post of operation,all patients got a significant amelioration in their symptoms. A total of 9 cases were followed up. There was 1 case of IIa type meningioma recurred. Conclusion To elevate the whole shearing rate of meningioma in skull base and reduce the mortality of it,it is most important for appropriate operative route selection and well mircosugical technique application.

Objective To establish a gas chromatography(GC) method for the determination of camphor,isoborneol,borneol,cinnamaldehyde and eugenol in Tonglikang ointment.Methods Polyethylene glycol with 100 %of application concentration was used as stationary liquid phase.GC was equipped with FID detector at 250 ℃and programmed heating method was applied.Results The resolution and the linearity were good with the recovery rate of camphor being 96.21 %(RSD=1.03 %),isoborneol 97.34 %(RSD=0.99 %),borneol 97.07 %(RSD=1.00 %),cinnamaldehyde 96.00 %(RSD=1.37 %),and eugenol 100.99 %(RSD=1.07 %) respectively.Conclusion The method is convenient,rapid and accurate.It can be used to control the quality of Tonglikang ointment.

The cDNA fragment of JEV prME gene was cloned into the baculovirus shuttle vector (bacmid) to construct a recombinant baculovirus vector, defined as AcBac-prME. Then the recombinant baculovirus Ac-prME was obtained by transfecting Sf9 cells with AcBac-prME. Western blot analysis and immunofluorescence results indicated that both prM and E proteins were efficiently expressed in Sf9 cells. Electron microscopy suggested that prME was assembled into JEV-VLPs. To further evaluate the potential of JEV-VLPs as vaccine, the mice were immunized with JEV-VLPs and then challenged with lethal JEV. The results of mice survival and pathological changes demonstrated that the JEV-VLPs performed complete protection against JEV-P3 strain and relieved pathological changes in the mice brain significant. This study suggest that JEV-VLPs would be a potential vaccine for Japanese encephalitis virus.
Animals ; Antibodies, Viral ; immunology ; Encephalitis Virus, Japanese ; genetics ; immunology ; Encephalitis, Japanese ; immunology ; prevention & control ; virology ; Humans ; Japanese Encephalitis Vaccines ; administration & dosage ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Sf9 Cells ; Vaccination ; Vaccines, Virus-Like Particle ; administration & dosage ; genetics ; immunology ; Viral Envelope Proteins ; administration & dosage ; genetics ; immunology

【Objective】 To verify the key performance parameters of Roche Cobas S201 multiplex NAT for blood donors, so as to provide data for the conformation of multiplex NAT performance and establish a scientific and feasible verification scheme. 【Methods】 Comprehensive performance verification on four key parameters of ROCHE COBAS S201 multiplex NAT system, including the 95% detection limit, compliance rate, anti-interference ability and operator comparison were conducted. The data were compared with the specifications provided by the manufacturer to evaluate the performance of laboratory testing system and ensure the quality of blood testing. 【Results】 2 sets of COBAS S201 multiplex NAT system in our laboratory were used to perform 5 times of the 95% detection limit value declared by the manufacturer for individual tests. The experimental results showed that the target NAT can be detected 100% and the lower limit of determination was verified. Single-virus NAT was performed to detect 5 different HBV DNA concentrations (25 ~400 IU/mL), 5 different HCV RNA concentrations (25 ~400 IU/mL), 5 different HIV RNA Concentration (100~1 000 IU/mL), and 5 negative tubes. The experimental results showed that all detected values of 20 tubes were 100% consistent with the true value, and the performance parameter "coincidence rate" was verified. Samples containing lipoemia, hemolysis, with ALT>50 mL/L and syphilis-positive endogenous interfering substances with 3 times of the 95% detection limit value were tested for 3 times, and results showed both response rate (3/3) and yielding rate reached 100%. In the control group (with none or in the normal range of interfering substances), reactivity was detected in samples with extremely low concentration of 3 times of the 95% detection limit, showing no significant difference (P>0.05). Above results fully showed that the existence of interference substances (lipidemia, hemolysis, ALT ≥ 50 and syphilis positive) in the blood of donors had no significant effect on the yielding of HBV/HCV/HIV virus target nucleic acids. Different operators performed the sample loading tests with same instrument, reagent and specimen showed 100% consistency in the results. It could be preliminarily assessed that there was no difference in the operations between the operators in this verification. 【Conclusion】 The verification scheme of the multiplex NAT system for methodology performance index (95% detection limit, coincidence rate, anti-interference ability, and operator comparison) established in this study, showed simple, flexible, scientific and feasible characteristics and could provide reference and data support for domestic blood transfusion services in acid detection.

Objective To modify the purified water analysis methods in the Chinese Pharmacopoeia (Vol .Ⅱ ,Edition 2010) ,and mainly focus on the pH value determination ,analysis of nitrate and nitrite ,and the inspection of microorganism . Methods The usage of regents and colorimetric tubes in the analysis of nitrate and nitrite were inquired ,the feasibility of plat-ing culture used in the inspection of microorganism were proposed ,and the suggestion of using more instrumental analysis in-stead of titrimetric analysis in the purified water inspection wasraised .Results The choice of colormetric apparatus and the precision of the regents were explicit .The plating method was better in convenience and reliability than membrane-filter proce-dure during the microorganism inspection .Instrumental analysis is the trend of purified water inspection and deserved to be rec-ommended .Conclusion Based on our own and others′studies ,the methods of impurity analysis are improved ,which might be accepted in the revision of Chinese Pharmacopoeia in future .

OBJECTIVE:To observe therapeutic efficacy and safety of cefoperazone sodium and sulbactam sodium combined with Shenmai injection in the treatment of elderly stroke associated pneumonia(SAP). METHODS:A total of 84 SAP patients in Dazhou Municipal Central Hospital during Mar. 2016-Mar. 2017 were divided into control group(42 cases)and observation group (42 cases)according to random number table. Based on routine treatment,control group was additionally given Cefoperazone sodium and sulbactam sodium for injection 3 g added into 5% Glucose injection 100 mL,intravenously,twice a day;suitable antibiotics were selected according to the results of pathogenic examination and drug sensitivity test. Observation group was additionally given Shenmai injection 100 mL added into 5% Glucose injection 250 mL intravenously,once a day,on the basis of control group. Both groups were treated for consecutive 2 weeks. Clinical efficacies of 2 groups were observed,and the levels of T-lymphocyte subsets(CD4+,CD8+,CD4+/CD8+)and inflammatory markers(TNF-α,hs-CRP,PCT),length of hospital stay before and after treatment. The ADR was recorded. RESULTS:The total response rate(95.24%)and marked response rate (47.62%)of observation group were significantly higher than those of control group(83.33%,23.81%);length of hospital stay in observation group [(15.24±3.53)d] was significantly shorter than control group [(18.43±4.21)d],with statistical significance (P<0.05).After treatment,the levels of CD4+and CD4+/CD8+in 2 groups were significantly higher than before treatment,and the observation group was significantly higher than the control group;the levels of CD8 +,TNF-α,hs-CRP and PCT in 2 groups were significantly lower than before treatment;the observation group was significantly lower than the control group,with statistical significance(P<0.05);there was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Based on routine treatment,cefoperazone sodium and sulbactam sodium combined with Shenmai injection treat SAP significantly,can effectively improve immune function, reduce inflammation level and shorten the length of hospital stay without increasing the occurrence of ADR.

Objective:To compare the clinical efficacy of transperitoneal robot assisted laparoscopic radical prostatectomy (RARP) versus extraperitoneal single port RARP.Methods:A retrospective analysis was perfoumed on 142 cases of RARP from July 2019 to June 2020 in Robotic Minimally Invasive Center of Sichuan Provincial People's Hospital, including 82 cases(Group A) , aged 70.0(65.6-78.0), undergoing transperitoneal RARP and 60 cases(Group B), aged 68.2 years old(60.1-79.2), undergoing extraperitoneal single port RARP. For group A, PSA was 12.9(5.6-64.0) ng/ml, with 26 cases of less than 10 ng/ml(31.7%), 40 cases of 10-20 ng/ml(48.8%), and 16 cases of more than 20 ng/ml(19.5%), the Gleason score was 7.2(6.0-10.0), with 14 cases(17.1%) of ≤6, 56 cases(68.3%) of 7, and 12 cases(14.6%) of ≥8, and the prostate volume was 61.3(29.0-112.0) ml. There were 49 cases with BMI≤25 kg/m 2, accounting for 59.8%, and 33 cases with BMI>25 kg/m 2, accounting for 40.2%. There were 17 cases(20.7%) of T 1, 44 cases(53.7%) of T 2 and 21 cases(25.6%) of T 3. The proportion of lymph node dissection was 17.1%, and 4 cases(4.9%)had a history of operation. For group B, the PSA was 12.2(1.0-42.6)ng/ml, with 20 cases (33.3%) of <10 ng/ml, 31 cases(51.7%)of 10-20 ng/ml, and 9 cases (15%) of >20 ng/ml. Gleason score was 7.1(6.0-9.0), with 12 cases (20.0%) of ≤6, 42 cases (70.0%) of 7, and 6 cases (10.0%)of ≥8. Prostate volume was 42.4(31.2-72.8)ml on average. There were 37 cases (61.7%) with BMI≤25 kg/m 2, and 23 cases (38.3%)with BMI >25 kg/m 2 . There were 17 cases(28.3%)of T 1, 32 cases(53.3%)of T 2 and 11 cases(18.3%)of T 3.The proportion of lymph node dissection was 11.7% and 4 cases (6.7%) had a history of operation.There was no statistically significant difference in term of age, PSA level, Gleason score, BMI, clinical stage, proportion of lymph node dissection or history of operation between the two groups( P>0.05), but there was statistically significant difference for prostate volume( P<0.05). All operations were performed by the same operator. Four different ways of bladder neck and urethral dissociation was selected according to the intraoperative conditions in Group A, include VIP style, T-shape incision style, VIP plus T-shape incision style or the style along the lateral side of the bladder neck. Small and single anterograde incision stripping of bladder neck was routinely performed in the Group B. Postoperative follow-up was performed to compare the operation time, intraoperative blood loss, bladder neck and urethral anastomosis time, postoperative hospital stay, postoperative exhaust time, postoperative complications, positive rate of surgical margin, indwelling time of urinary catheter, urinary continence satisfaction rate of immediately after operation, 3 months and 6 months after operation, wound healing and aesthetics. Results:All of the operations were successfully completed under robot-assisted laparoscopy, and there was no conversion to open surgery. The operation time was 56.0(45.0-112.0) min in the Group A and 65.4(55.5-96.8) min in the Group B, and there was no statistically significant difference( P>0.05). The intraoperative blood loss was 76.2(30.0-120.5) ml and 55.6(45.5-114.6) ml, respectively, and the difference was not statistically significant( P>0.05). The time of bladder neck urethral anastomosis was 18.9(12.6-25.6) min and 16.2(10.7-19.3) min, respectively, and the difference was not statistically significant( P>0.05). The postoperative hospital stay days were 9.3(8.0-16.0) d and 8.4(7.0-13.0) d, respectively, and the difference was not statistically significant( P>0.05). The postoperative exhaust time was 1.3(0.7-3.0) d and 3.4(2.0-7.0) d, respectively, and the difference was statistically significant( P<0.05). There was 1 case of anastomotic fistula with ureteral injury in Group A, and no serious complication in Group B, and the difference was not statistically significant( P>0.05). The number of positive surgical margin in the two groups was 13(15.9%)and 9(15.0%)respectively, and the difference was not statistically significant( P>0.05). The indwelling time of urinary catheter after operation was 9(7-21) d and 6(4-8) d, respectively, and the difference was statistically significant( P<0.05). The number of patients with satisfactory urinary continence immediately after surgery, 3 months and 6 months after surgery in the two groups were 8(9.8%), 51(62.2%), 62(75.6%) and 17(28.3%), 43(71.7%) and 54(90.0%), respectively. The differences were statistically significant( P<0.05). The total incision lengths in the two groups were 12.1(10.4-13.4) cm and 5.6(5.0-6.0) cm, respectively, and the difference was statistically significant( P<0.05). Conclusions:The extraperitoneal single port RARP is safe and feasible, and the postoperative effect is similar to that of transperitoneal RARP. It has the advantages of shorter recovery time, higher urinary continence satisfaction rate, neater and more beautiful incision. The long-term therapeutic effect needs further confirming by prospective study.

Six patients with prostate cancer, treated by suprapubic extraperitoneal single-port robot-assisted radical prostatectomy, had been studied retrospectively from December 2019 to January 2020. All 6 patients have been treated by suprapubic peritoneum single port robot assisted laparoscopic surgery without other channels. The robot assisted laparoscopic radical prostatectomy via suprapubic peritoneum is safe and feasible when based on reasonable selection criteria of patients. It has been shown that the postoperative recovery was fast and the tumor control and continence rate were good under the short-term follow-up. However, the long-term outcome should be evaluated by a long-term follow-up.