Objective To investigate the regulatory effect of small interfering RNA(siRNA)silencing zinc finger protein A20 on pyroptosis of rheumatoid arthritis(RA)fibroblast-like synoviocytes(HFLS-RA).Methods Hu-man FLS-RA cell line MH7A cells were cultivated,A20 siRNA silencing group was synthesized for knocking down the human A20 gene,and then specific A20 gene siRNA and siRNA-NC(negative control)were transfected into MH7A cells using liposome method.RT-qPCR was applied to detect the expression of NLRP3 and Caspase-1 mRNA in cells.The protein expression of NLRP3 and Caspase-1 was detected by Western blot,and IL-1β and IL-18 in cell culture medium were detected by ELISA method.Transmission electron microscopy(TEM)was used to detect pyroptosis.Results After A20 knockdown,the mRNA and expression of NLRP3 and Caspase-1 in MH7A cells in the siRNA-A20 group were significantly increased as compared with the siRNA-NC group(P＜0.01).The concentration of IL-1β and IL-18 in the cell culture supernatant of the siRNA-A20 group was sig-nificantly increased compared with the siRNA-NC group(P＜0.01).Compared with the siRNA-NC group,some cells in the siRNA-A20 group showed swollen and ruptured.The integrity of the cell membrane was also lost,and a large area of edema was present in the cell.In addition,a blurred depression of the local nuclear membrane was noted,while an increase in heterochromatin pyknosis was accompanied by their uneven distribution as well as their aggregation around the nuclear membrane.Conclusions Silencing of A20 gene with siRNA might promote NLRP3/Caspase-1 mediated pyroptosis in HFLS-RA,which lays an experimental foundation for new clinical treatment meth-ods of RA.

Objective To explore method of establishing and evaluating an asthma rat model with phlegm and blood stasis syndrome.Methods 60 SD male rats were randomly divided into 5 groups,a normal group,asthma group,combination of disease and syndrome(combination)group,DM group,and KCLW group,with 12 rats in each group.Asthma models were established using ovalbumin(OVA).A syndrome model of phlegm and blood stasis was established using a high-fat diet combined with the ice water bath method.We evaluated the asthma model through animal behavior observation,pathological section observation,inflammation index detection,and airway reactivity measurements.The phlegm and blood stasis syndrome model was evaluated via measurements of rat body mass,blood glucose,blood lipids,coagulation function,and hemorheological indexes and by observing symptoms and syndrome determination by Kechuan Liuwei mixture.Results(1)After OVA induction,the rats in the asthma model group and combination group showed symptoms such as shortness of breath,open mouth breathing,abdominal movement,restlessness,and irritability.HE staining showed the disordered arrangement of the bronchial mucosa in lung tissue,local detachment,thickening of the basement membrane and the bronchial tube wall,narrowing of the lumen,extensive infiltration of inflammatory cells,and congestion of capillaries.Compared with the normal group,the asthma model group and combination group(P＜0.05)had increased serum IL-4,IL-6,and TGF-β1.Penh values were increased after stimulation with various concentrations of Mch(P＜0.05).(2)Rats in the combination group showed symptoms such as chills,curling up with minimal movement,purple and dark claws,purple and black bruises on the tail,loose stools,and unclean perianal area.Compared with the rats in the asthma model group,rats in the combination group had increased body mass(P＜0.05)and blood glucose,triglyceride,and total cholesterol levels(P＜0.05),a shortened thrombin time(P＜0.05),increased fibrinogen content(P＜0.05),and significantly increased whole-blood viscosity at low,medium,and high shear rates(P＜0.05).The indexes were significantly improved after Kechuan Liuwei mixture administration.Conclusions The asthma rat model with phlegm and blood stasis syndrome can be established through OVA induction and high-fat diet combined with ice water bath.The model can be evaluated through behavioral observation,index measurements,and syndrome determination via formulas.

Objective:To investigate the safety, economic effect, and short-term clinical efficacy of ambulatory total hip arthroplasty (THA) in the treatment of hip osteopathy within.Methods:This study retrospectively reviewed patients who underwent 48-hour outpatient THA and conventional primary THA from July 2020 to July 2021. Gender, age, body mass index (BMI), clinical diagnosis, place of resident, length of hospital stay, duration of the surgery, hemoglobin, albumin, C-reactive protein (CRP) and D-dimer before and 1 day after surgery, the visual analogue scale (VAS) was obtained postoperative day 2, hip joint modified Harris score before and 1 month after surgery, readmission and reoperation within 1 month after operation as the data of evaluations were extracted and compared in this study.Results:A totally of 150 cases were involved in this study, which including 75 cases with 48-hour outpatient primary THA and 75 cases with conventional primary THA. There was no significant difference between the two groups in terms of age, sex, BMI, education level, preoperative diagnosis, and preoperative Harris score ( P>0.05), but a significant difference was found in term of residence (χ 2=6.29, P=0.043), that the patients in the outpatient group were all from Zhejiang Province and 48% (36/75) of them were from Hangzhou City. While, in the conventional group, 6 patients were from other provinces. The length of stay was 2.13±0.52 days and operation time was 59.73±18.91 minutes in the outpatient group, which were both shorter than those (6.71±1.44 days and 66.91±22.40 min) in the conventional group ( t=25.91, P<0.001 for the length of hospital stay; t=2.12, P=0.036 for operation time). Compared with the conventional group, outpatient group saved the average hospital cost (4.60±0.44 vs. 5.20±0.72 ten thousand yuan, t=6.16, P<0.001). The VAS on the second day after surgery 3.45±0.75 was higher in the outpatient group than in the conventional group (3.45±0.75 vs. 3.16±0.94 points, t=2.09, P=0.039). The modified Harris score was without statistical significance ( t=0.42, P=0.677) 1 month after surgery in both groups. 75 patients in the outpatient group, 7 patients delayed discharge (were not discharge within 48 hours), and the rate of delayed discharge was 9.3%. Reasons for delayed discharge included poor pain control in two cases, one case had postoperative nausea and vomiting, one case had failed to meet rehabilitation standards, hypotension in one case, the intraoperative infection in one case and postoperative fever within 48 hours in one case. Conclusion:Outpatient THA can reduce the length of hospital stay, operative time and total cost of hospitalization. It has similar safety and early clinical efficacy as conventional THA. However, a small proportion of patients would delay discharge.

Objective:To evaluate the impacts of tissue heterogeneity on dose calculation of cervical brachytherapy by comparing the doses calculated by two clinically used dose calculation method and the CT image-based Monte Carlo (MC) method.Methods:This study retrospectively selected 11 patients with cervical cancer treated with 3D brachytherapy in Anhui Provincial Cancer Hospital from January 2018 to June 2020. The dose distribution of each plan was calculated via three methods, dose calculation method described in American Association of Physicist in Medicine(AAPM) Task Group No. 43 Report (TG43-BT), Acuros BV(BV-BT) used to perform accurate dose calculations in high-dose-rate (HDR) brachytherapy with phantom heterogeneity, and CT image-based EGSnrc tool kit used to perform Monte Carlosimulation (MC-BT). The dose volumes( V3 Gy, V6 Gy, V9 Gy, and V12 Gy), target volume doses( D98, D90, D50), D2 cm 3 of organs at risk (OARs) calculated by the three methods were compared. Results:The HRCTV D90obtained by TG43-BT was 6.274 Gy, which was even overestimated by around 5% compared to the result calculated by MC-BT. Meanwhile, TG43-BT overestimated the dose volumesand the target volume doses compared to MC-BT.Except for D50 and V12 Gy, the differences between the doses to tumor calculated by BV-BT and MC-BT were not statistically significant( P>0.05). There was also no significant statistical difference between the D2 cm 3 of rectum, small intestine, and sigmoid calculated by BV-BT and MC-BT ( P>0.05). In contrast, the dose to D2 cm 3 of bladder determined by MC-BT was 4.609 Gy, which was notably higher than those deter mined by TG43-BT and BV-BT. Conclusions:TG43-BT overestimated the doses to tumor targets and most OARs since the effects of tissue heterogeneity were not taken into consideration. BV-BT performed efficient calculation and most of the dose distributionin target volume and OARs obtained by BV-BT were consistent with that calculated by MC-BT. Nevertheless, low accuracy occurred for the regions near the sources and full bladder, which warrants further caution in clinical evaluation.

Objective:To investigate the impacts of the composition and physical density of tissue on the dose distribution of implanted 125I seeds, in order to provide references for the clinical dose calculation and assessment of implanted radioactive particles. Methods:The OncoSeed 6711 physical model of 125I seeds was established using thes of twareegs_brachy and was validated through the calculation of dose rate constant and the radial dose function [ g( r)] in water. Then, based on the element composition and physical density of different types of tissue, the g( r) and absorbed dose ratein water, prostate, breast, muscle, and bone were calculated. Results:The calculated dose rate constant (0.950 cGy·h -1·U -1) and g( r)in water approached the values in related literature. The absorbed dose in bone was 6.042 times than that in water at a distance of 0.05 cm from the implanted source. The difference between the absorbed doses in breast and water was more than 10% at a distance of less than 1.7 cm from the implanted source. The difference between the absorbed doses in prostate/muscle and water was less than 5% at the same radial location. Conclusions:The dose distribution of 125I seeds in some types of human tissue is significantly different from that in water, which should be carefully considered in clinical dose calculation.

OBJECTIVE： To study the effects and mechanism of Le’ermai capsule on oxidative stress injury in the cerebral cortex of cerebral ischemia-reperfusion injury rats. METHODS： Totally 65 rats were randomly divided into sham operation group （normal saline）， model group （normal saline）， positive control group （Naoxintong capsule， 3.40 g/kg）， Le’ermai capsule high-dose and low-dose groups （0.37， 0.18 g/kg）， with 13 rats in each group. Except that sham operation group received sham operation， and middle cerebral artery occlusion/reperfusion （MCAO/R） model was induced by suture-occluded method. 24 h after modeling， they were given relevant medicine intragastrically， once a day， for consecutive 7 d. 2 h after last administration， cerebral ischemic area was determined by TTC staining. The pathological changes of cerebral tissue were observed by HE staining. The protein expressions of HO-1， SOD1， SOD2 and Nrf2 in cerebral cortex were detected by Western blot assay. RESULTS： Compared with sham operation group， the area of cerebral ischemic was increased significantly in model group（P＜0.05）， interstitial edema was serious， inflammatory cell infiltration increased significantly； and the protein expression levels of HO-1 and SOD1 in cortex tissue were significantly increased （P＜0.05）， and the protein expression levels of SOD2 and Nrf2 were significantly decreased （P＜0.05）. Compared with model group， the area of cerebral ischemic area of Le’ermai capsule high-dose and low-dose groups were significantly decreased （P＜0.05）. The interstitial edema， inflammatory cell infiltration and microcytes proliferation were decreased. The protein expression levels of HO-1， SOD1， SOD2 and Nrf2 in the cerebral cortex tissue were significantly increased （P＜0.05）. CONCLUSIONS：Le’ermai capsule can improve cerebral ischemia/reperfusion injury to certain extent， and which may be associated with up-regulating the protein expression of antioxidant factors HO-1， SOD1， SOD2 and Nrf2 in the cortex.

Objective: To analyze many indicators during perioperative period of liver transplantation in patients with end-stage liver disease, only to seek related factors that can accurately predict short-term(≤three months)outcome. Methods: We analyzed retrospectively clinical data of consecutive patients with end-stage liver diseases undergone liver transplantation in a single treatment center. Logistic regression analysis was used to analyze the perioperative indicators including recipient gender, age, body mass index, preoperative serum albumin level, serum sodium concentration, urea nitrogen level and donor-recipient blood group, et al. Correlated factors were analyzed by the method of multivariate logistic regression. Statistical processing package was SAS 9.1.3 soft. The difference was statistically significant with P<0.05. Results: 18/165 patients died within 3 months after transplantation(mortality rate: 10.9 %). According to the result of univariate analysis, the indicators correlated with early mortality which were statistically significant were preoperative serum sodium, blood urea nitrogen, PT-INR, CTP score, MELD score and MELD-Na score. On the base of the result of Logistic multiple regression. However, only MELD-Na score was associated with 3 months prognosis(P=0.001, β=-2.510, OR=0.088, 95 % CI=0.037~0.349). Conclusions Preoperative MELD-Na score is an independent risk factor for short-term survival in patients with end-stage liver disease. Higher MELD-Na score is, the early mortality is higher.

BACKGROUND AND OBJECTIVES: The International Society for Cellular Therapy (ISCT) proposed a set of minimal markers for identifying human mesenchymal stromal cells (hMSCs) in 2007. Since then, with the growing interest of better characterising hMSCs, various additional surface markers have been proposed. However, the impact of how culture conditions, in particular, the culture surface, vary the expression of hMSC markers was overlooked. METHODS AND RESULTS: In this study, we utilized the RNA sequencing data on hMSCs cultured on different surfaces to investigate the variation of the proposed hMSC biomarkers. One of the three ISCT proposed positive biomarker, CD90 was found to be significantly down regulated on hMSCs culture on fibrous surfaces when compared to flat surfaces. The detected gene expression values for 177 hMSCs biomarkers compiled from the literature are reported here. Correlation and cluster analysis revealed the existence of different biomarker communities that displayed a similar expression profile. We found a list of hMSCs biomarkers which are the least sensitive to a change in surface properties and another list of biomarkers which are found to have high sensitivity to a change in surface properties. CONCLUSIONS: This study demonstrated that substrate properties have paramount effect on altering the expressions of hMSCs biomarkers and the proposed list of substrate-stable and substrate-sensitive biomarkers would better assist in the population characterisation. However, proteomic level analysis would be essential to confirm the observations noted.
Biomarkers ; Chemistry ; Gene Expression ; Humans ; Mesenchymal Stromal Cells ; Quality Control ; Regenerative Medicine ; Sequence Analysis, RNA ; Surface Properties ; Transcriptome

Objective To investigate the effect of sodium houttuyfonate on the expression of PI3K and AKT1 and mTOR mRNA in the lung of rats with chronic obstructive pulmonary disease(COPD),and reveal the possible mechanism of the COPD treated with sodium houttuyfonate. Methods Twenty-four male Wistar rats were randomly divided into normal control group,model control group,dexamethasone group and sodium houttuyfonate group(n=6 for each). The rat models of COPD were established by intratracheal instillation of lipopolysaccharide and smudging. The expressions of PI3K and AKT1 and mTOR mRNA were determined by real-time PCR. The morphological changes of the lung tissue was examined by histopathology. Results Compared with the normal control group,the expressions of PI3K and AKT1 were significantly in-creased and mTOR mRNA was significantly decreased in the model group(P<0.01,P<0.05). Compared with the mod-el group,the expressions of PI3K and AKT1 were significantly decreased and mTOR mRNA was significantly increased in the sodium houttuyfonate group and dexamethasone group(P<0.01,P<0.05). Compared with the dexamethasone group, the expression of mTOR mRNA was significantly increased in the sodium houttuyfonate group(P<0.05). The pathological observation indicated that there were local pulmonary consolidation and a extensive neutrophil infiltration in the alveolar cav-ity. Prominent pulmonary interstitial fibrous hyperplasia was observed in the model group. The pathological manifestations were much ameliorated than those of the model group,and only mild interstitial pneumonia and a slight fibrous hyperplasia were seen in the sodium houttuyfonate and the dexamethasone groups. Conclusions Sodium houttuyfonate reduces the in-jury of lung tissue and has protective effect on COPD rats. The mechanism is probably related to the down-regulatation of expression of PI3K and AKT1 mRNA and up-regulatation of expression of mTOR mRNA in COPD rats.

OBJECTIVE:To investigate clinical efficacy and safety of linezolid in the treatment of intracranial infection after neurosurgery operation. METHODS:Medical information of 39 intracranial infection patients receiving linezolid in Xijing Hospital of Air Force Medical University during Jul. 1st,2015-Aug. 29th,2016 were analyzed retrospectively. The clinical efficacy andsafety of linezolid in the treatment of intracranial infection after neurosurgery operation were evaluated according to indexes of intracranial infection patient,such as symptoms,signs,lab indexes test and bacterial culture results. RESULTS:Total response rate of 39 intracranial infection patients after neurosurgery operation was 79.49％ after linezolid treatment. After treatment,the patients' body temperature,white blood cell,neutrophil absolute value,white blood cell in cerebrospinal fluid and cerebrospinal fluid protein level were all significantly lower than before treatment,with statistical significance(P<0.05). Of 39 patients,cerebrospinal fluid of 27 patients were cultured before treatment,and 8 cases(29.6％)of which were positive,among which there were 6 cases (75.0％) of Gram-positive bacteria such as Staphylococcus and Enterococcus. No obvious ADR related to linezolid was found in patients. CONCLUSIONS:Linezolid can effectively control the intracranial infection caused by Gram-positive bacteria such as Staphylococcus and Enterococcus with good safety.