[Objective] To observe the effect of stanozolol on proliferation and differentiation of cultured growth plate chondrocytes in vitro.[Methods] At 3 week of age,Sprague–Dawley rats received 2.5 mg/kg in slow-released GnRHa (triptorelin) which was repeated every 2 weeks for 2 times,at 7-week old.The tibial growth plate cartilage were aseptically dissected and tripsin and EDTA digested for 0.5 h,then collagenase digested for 3 h at 37 ℃.Chondrocytes were cultured in DMEM:F12 medium for 48 h,the cells were starved for 24 h in serum-free DMEM:F12 medium before stanozolol treatment.In dose-effect groups,chondrocytes were incubated in serum-free media in various concentrations of stanozolol for 48 h.In time-course groups,chondrocytes were incubated in serum-free media in various times of stanozolol (10-8 mol/L).immunohistochemical staining of collagen Ⅱ,Ⅹ,PCNA,and MTT were conducted.[Results] The results of MTT,PCNA,and typeⅡcollagen synthesization demonstrated stanozolol enhanced the proliferation of the chondrocytes,time-course studies had shown that the proliferation were maximally stimulated by stanozolol after 2 or 3 days of incubation and decreased again after longer periods of incubation.Stanozolol stimulated the proliferation of the chondrocytes dose-dependently at 10-11 mol/L and 10-8 mol/L,maximally stimulatory concentrations of Stanozolol was 10-9 ～ 10-8 mol/L,and decreased again after higher concentration of stanozolol.Stanozolol did not stimulated type X collagen synthesization from 10-11 mol/L ～ 10-8 mol/L,but experiments showed that type X collagen was already stimulated after incubation in Stanozolol (10-7 ～ 10-5 mol/L).Time-course studies had shown those typeⅩcollagen synthesizations were stimulated by stanozolol after 4 ～ 5 days of incubation.[Conclusion] Stanozolol enhances the proliferation of chondrocytes of pubertal female rat treated with GnRHa in vitro (time-course- dependent and concentration -dependent).

Objective To explore the inhibitive effect of BSP on Caspase dependent apoptosis of preosteoclasts RAW264.7 cells by bone sialoprotein (BSP).Methods RAW264.7 cells were treated with the inhibitors of signaling pathway molecules and/or BSP.The activities of caspase3/8/9 in RAW264.7 cells were detected with caspase3/7/8/9 kits and the interaction between Caspase-8 and integrin αVβ3 was detected by Co-IP.Results The activities of Caspase3/8/9 in BSP-treated RAW264.7 cells were decreased and the apoptosis of RAW264.7 cells were inhibited significantly.In addition,there was a positive correlation between caspase-8 activity and caspase-3 activity.There was interaction between caspase-8 and integrin αVβ3.Conclusion The anti-apoptotic effect of BSP on RAW264.7 cells is mainly through the regulation of exogenous apoptosis pathway and the apoptosis induced by caspase-8 is inhibited by the binding of BSP and αvβ3.

Objective To investigate the expression of miR-520d-3p in 2 and 4 Gy X-ray irradiated A549 cells, serum samples of non-small cell lung cancer (NSCLC) patients and its significance. Methods Real time quantitative PCR (RT-qPCR) was employed to detect the expression miR-520d-3p in 2 and 4 Gy X-ray irradiated A549 cells in vitro. Besides, 0, 2 and 4 Gy X-ray irradiated A549 cells were used to prepare animal model of nude mice lung metastasis through mainline. The expressions of miR-520d-3p and lung tissue of nude mice were detected, and the serum samples of NSCLC patients were collected to test the expression of miR-520d-3p . Results Compared with the control group ( 0 Gy ) , the expression of miR-520d-3p was up-regulated significantly in 2 and 4 Gy X-ray irradiated A549 cells after 48 hours (1.00±0.03, 1.47±0.10, 1.84 ±0.09 respectively), and there was a significant difference (F= 94.350, P= 0.000). Furthermore, compared the control group with injection after 10 weeks, the expressions of miR-520d-3p in nude mice lung tissue in 0, 2 and 4 Gy X-ray irradiated groups were increased (2.33 ±0.13, 8.24 ±0.25, 3.46 ±0.14, respectively) (F=1.787, P= 0.227), and the expressions in serum were increased (11.43±2.30, 10.22±4.62, 8.99 ±3.67, respectively) (F= 1.547, P= 0.246). Out of 20 serum samples of NSCLC patients (including 10 cases of adenocarcinoma, 10 cases of squamous carcinoma ), 11 cases (55%) were detected with up-regulated miR-520d-3p expression. Conclusions 2 and 4 Gy X-ray could increase the expression of miR-520d-3p of A549 cells in vitro and in vivo, which might be correlated with the enhanced invasion and metastasis of A549 cells induced by X-ray in vitro and in vivo. Furthermore, the expressions of miR-520d-3p were increased significantly in over 50%serum samples of NSCLC patients, which might be a new marker for the diagnosis of NSCLC.

Objective To establish a stem cell life-cycle management information system to realize collection,storage,inquiry,processing and statistical analysis of the stem cell and to provide information for the research on stem cell translation medicine.Methods The functional modules of the system were determined based on the analyses on the requirements of stem cell bank management and its operational flow.The system design was explored from the aspects of technical framework,safety control and fuzzy query.Results The system was gifted with a technical architecture and anallocation scheme based on Spring MVC mode,which proved its efficiency by the trial.Conclusion Stem cell resource library can be used as an important supplement to the clinical sample life-group database,providing data and technical basis support for large-scale clinical samples to transform,integrate,manage and share large data resources.

Objective To explore the key signaling pathways and molecules of bone sialoprotein (BSP) to promote proliferation and differentiation of preosteoclasts RAW264.7. Methods RAW264.7 cells were treated with BSP and the inhibitors of signaling pathway molecules. MTS and TRAP staining kits were used to evaluate the effects of proliferation and differentiation. The activity assay kits of Calcineurin, AKT, JNK, ERK and p38 were used to detect their activities. Results Inhibiting ERK and p38 can inhibit cell proliferation induced by BSP significantly, while inhibiting AKT, Calcineurin and PI3K can reduce the role of promoting the differentiation by BSP. With increasing treatment time of BSP, the activity of Calcineurin in RAW264.7 cells increased. Furthermore, the activity of AKT was maximum at 48 h after treated with BSP, while the activity of JNK, ERK and p38 were maximum at 24 h after treated with BSP. Conclusions BSP mainly regulates the ERK and p38 of MAPK pathway to promote RAW264.7 cell proliferation, and regulates AKT, PI3K and Calcineurin pathways to promote RAW264.7 cell differentiation.

Objective To investigate the expression of miR-424* in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of non-small cell lung cancer(NSCLC) patients,and to explore its potential role in the diagnosis and prognosis of lung cancer.Methods A549 cells were irradiated with 2 and 4 Gy X-rays,and some of irradiated cells were injected into nude mice through tail vein.Real time quantitative PCR (RT-qPCR) assay was employed to detect the expression of miR-424 * in 2 and 4 Gy X-ray irradiated A549 cells in vitro and in vivo,as well as in clinical lung tissues and serum sample of lung cancer patients.Results Compared with the control group,the expression of miR-424* was up-regulated significantly in X-ray irradiated A549 cells at 1,2,12,24 and 48 hpost irradiation,respectively (2 Gy:t =-45.886--6.709,P ＜0.05;4 Gy:t =-29.087--7.833,P ＜ 0.05).Furthermore,the expression of miR-424 * was up-regulated in the lung and serum of nude mice with injection of 0,2 and 4 Gy X-ray irradiated A549 cells,compared with control group (fold change was 9.72,8.58 and 4.7 with 2 Gy irradiation and 11.93,9.22 and 8.99 with 4 Gy irradiation,t=-13.243,-12.409,-9.833 in lung andt=-6.436,-3.052,-3.609 in serum,respectively,P ＜ 0.05).Out of 11 tissue samples of NSCLC patients,6 were detected with up-regulated miR-424* expression,and no significant discrepancy of miR-424* expression was detected in two type of NSCLC tissue samples.On the contrary,43 serum samples were detected with up-regulated miR-424* expression out of 84 serum samples (51.20％) of NSCLC patients (fold change range 1.97 to 17.71),and significant discrepancy of miR-424* expression was shown in two subtypes of NSCLC serum samples [adenocarcinoma:39.10％ (18/46) and squamous carcinoma:65.8％ (25/38)],as well as in serum samples of NSCLC patients with radiotherapy [41.5％ (22/53)] and without radiotherapy [67.7％ (21/31)] (t=5.919,5.387,P ＜0.05,respectively).Conclusions 2 and 4 Gy X-ray irradiation could up-regulate the expression of miR-424* in A549 cells,which might be correlated with the enhanced metastasis of A549 cells induced by X-ray in vivo and in vitro.Furthermore,the expression of miR-424* was up-regulated in over 50％ of the tissue and serum samples of NSCLC patients,which might be correlated with the diagnosis of NSCLC subtype and prognosis of radiotherapy.

Objective: To improve the opening method of humidifier, enhance the efficiency and success rate of opening the humidifiers. Methods: Use all the recycled humidifier from respirators in the wards from 1st of May to 31st of October 2015 as control group. Use the recycled humidifiers from 1st Feb to 31st July 2016 as the test group. Use metal tools to open humidifier bottles in the control group while use the newly developed and improved opening device to open the bottles in the test group to test and compare the damage rate. Results: The testing group that used the practical opening device for humidifier has a decrease in damage rate from 5.97% (16/268) to 1.22% (3/245), and this difference before and after was statistically significant (χ²=8.082, P<0.01). Conclusions The use of the opening device for humidifier can enhance the intact rate, greatly improve working efficiency and save the cost.

Objective:This study is to investigate the predictive value of serum levels of TIMP-2 and insulin-like growth factor-binding protein 7(IGFBP7) in patients with DCD(donation after cardiac death) kidney transplantation.Methods:A prospective research design was used to select DCD kidney transplant patients admitted to the Li Huili Hospital of Ningbo University from January 2018 to October 2020.Inclusion criteria: ①Complete data; ②There were no serious complications affecting the function of the transplanted kidney in the early postoperative period.Exclusion criteria: ①Incomplete data; ②Patients were unable or unwilling to cooperate with the study; ③Severe complications affecting the function of the transplanted kidney occurred early after the operation.The ELASE method was used to quantitatively detect the serum TIMP-2 and IGFBP7 levels at 6, 12, 24, 48, 72 hours and 7 days after renal transplantation, and monitor the serum creatinine values during the same period and 21 days after the operation. According to the occurrence of DGF, the measured values of TIMP-2 and IGFBP7 at different time points and their product's ability to predict the occurrence of DGF after kidney transplantation were analyzed. The receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to evaluate the diagnostic efficacy of TIMP-2 and IGFBP7 for DGF.Results:A total of 33 patients were enrolled, 7 patients (21.2%) in the DGF group and 26 patients (78.8%) in the non-DGF group. Between the two groups, the donor glomerular filtration rate were [98.5(15.8-132.5)ml/(min·1.73m 2) and 79.1(60.6-102.5)ml/(min·1.73m 2)], recipient gender (male/female: 3/4 cases and 10/16 cases), recipient age [48(34-56) Years old and 45(23-61) years old], the recipient's preoperative creatinine [1114.0(731.4-1293.0)μmol/L and 858.4(657.6-1051.9)μmol/L], the recipient's preoperative urea nitrogen [15.0(13.2-19.6)mmol/L and 17.3(13.6-20.9)mmol/L], receptor preoperative albumin [43.5(38.5-45.3)mmol/L and 41.2(37.5-46.1) mmol/L], recipient dialysis method [hemodialysis/peritoneal dialysis: 3/4 cases and 9/17 cases], warm ischemia time [6(5-7) and 5(4-6) min, there was no statistically significant difference] ( P>0.05). The values of serum IGFBP7 and TIMP-2×IGFBP7 in the DGF group were higher than those in the non-DGF group at all time points ( F=15.753, P=0.040; F=13.000, P=0.024), while serum TIMP-2 was not significant between the two groups difference ( F=1.157, P=0.075). For the diagnostic value of DGF, the AUC of serum IGFBP7 at 48 h after surgery was 0.863 (95% CI 0.696-1.000, P=0.004). When 5.97 ng/ml was used as the cut-off value, the sensitivity was 85.7% and the specificity was 80.8 %. The AUC of TIMP-2×IGFBP7 at 48 hours after surgery was 0.819 (95% CI 0.641-0.996, P=0.011). When 62.06(ng/ml) 2 was used as the cutoff value, the sensitivity was 71.4% and the specificity was 80.8%.There was no statistical difference in the area under the curve between the two ( P>0.05). There were differences in the dynamic trend of serum IGFBP7 and creatinine in the DGF group. Serum IGFBP7 at 7 days after surgery was positively correlated with creatinine at 21 days after surgery. Conclusion:Serum IGFBP7 and TIMP-2×IGFBP7 could predict the occurrence of DGF after DCD donor kidney surgery. The predictive value changes with time. Among them, 48h and 7d after surgery are the most valuable. However, serum TIMP-2 has not been found to have predictive value in this study.

Body temperature is an essential physiological parameter. Conducting non-contact, fast and accurate measurement of temperature is increasing important under the background of COVID-19. The study introduces an infrared temperature measurement system based on the thermopile infrared temperature sensor ZTP-135SR. Extracting original temperature date of sensor, post-amplification and filter processing have been performed to ensure accuracy of the system. In addition, the temperature data of environmental compensation which obtained by polynomial fitting is added to the system to further improve measurement accuracy.
Algorithms ; Body Temperature ; COVID-19 ; Humans ; Temperature ; Thermometers

Oxygen therapy is an effective clinical method for the treatment of respiratory disorders, oxygen concentrator as a necessary medical auxiliary equipment in hospitals, its research and development has been a hot spot. The study reviewed the development history of the ventilator, introduced the two preparation technique of the oxygen generator pressure swing absorption (PSA) and vacuum pressure swing adsorption (VPSA), and analyzed the core technology development of the oxygen generator. In addition, the study compared some major brands of oxygen concentrators on the market and prospected the development trend of oxygen concentrators.
Oxygen ; Oxygen Inhalation Therapy ; Hospitals ; Ventilators, Mechanical ; Equipment Design