1.KAT7 promotes the aging of cartilage cellular senescence
Xulei WANG ; Zhuping CHU ; Huimin WANG ; Wei WEI ; Shangxue YAN
Acta Universitatis Medicinalis Anhui 2024;59(2):293-297
		                        		
		                        			
		                        			Objective To investigate the role of KAT7 in cartilage cell and tissue ageing by establishing an over-replicating-induced primary mouse cartilage cell ageing model and a mouse natural ageing model.Methods Chon-drocytes of the mouse knee joint were obtained by type Ⅱ collagenase digestion and identified by toluidine blue stai-ning and Col Ⅱ staining.The age-related proteins and KAT7 expression levels in cartilage cells from different gener-ations of mice were discovered using Western blot and cellular immunofluorescence techniques,and the aging of the cells was assessed by SA-β-Gal coloring.The pathological alterations were examined in the joints of 22-month-old mice compared to 2-month-old mice using HE staining and safranin O-solid green staining.Additionally,immuno-histochemical analysis was done to observe the expression of KAT7 and p53 in mouse joint tissue.Results Com-pared with the control group,the expression levels of KAT7 protein and p21 and p53 in aged mouse chondrocytes significantly increased.WM-3835,a commonly inhibitor of KAT7 that possess the capacity on halting the protein expression procedure of gene KAT7 as well as p21 in ageing chondrocytes.SA-β-Gal staining showed a significant increase in positive staining of chondrocytes in the eighth generation(P8)compared to the first generation(P1).Compared with the cartilage tissue of young mice,the cartilage tissue of elderly mice presents a near-bone distribu-tion,with a decrease in cartilage surface integrity,a significant increase in the number of hypertrophic chondro-cytes,and more KAT7 and p53 cells that were positive.Conclusion The expression of KAT7 increases in the ageing chondrocytes and the cartilage tissue of ageing mice,reveales the potential significance of KAT7 correlated to cellular aging process in cartilage.
		                        		
		                        		
		                        		
		                        	
2.Effect of low-dose dexamethasone combined with PB21 on analgesic effect in rats with knee osteoarthritis
Zhuping CHU ; Tianxi DU ; Qiongxia XIE ; Xulei WANG ; Huimin WANG ; Xiaorong LU ; Shangxue YAN
Acta Universitatis Medicinalis Anhui 2024;59(7):1225-1230
		                        		
		                        			
		                        			Objective To examine the impact and partial mechanism of bupivacaine sustained-release drug(code PB21)in combination with low-dose dexamethasone(Dex)on the analgesic time of rats with knee osteoarthritis(KOA).Methods Using the techniques of anterior cruciate ligament transection and meniscus instability,a rat KOA model was created.After eight weeks,SD mice were split into three groups at random:a group for the model,one for Dex(50 μg),one for PB21(1.5 mg),and one for combined administration(1.5 mg PB21/50 μg Dex),with a control group that received a sham operation.The pain thresholds of KOA rats were measured using a Pres-sure Application Measurement(PAM)at different intervals before to delivery and 4,24,36,and 48 hours following administration;to gauge changes in discomfort,a CatWalk was used to assess the rats'average foot strength and maximum contact area before,four,twenty-four,and forty-eight hours after treatment.A portion of the rats were put to sleep at four,twenty-four,and forty-eight hours following the injection,and the joint synovium was removed for paraffin sectioning.Immunohistochemistry was used to identify the expression of GAP43 in the synovium,whereas immunofluorescence was used to identify the expression of CGRP in the same tissue.Results The average strength and maximum contact area of the foot and claw decreased(P<0.01),and the pain threshold decreased(P<0.01)in the model group compared to the sham operation group.The PB21+Dex group experienced a delayed pain threshold lowering time delay when compared to the PB21 and Dex treatment groups alone.Up to 48 hours lat-er,the combination administration group's average strength and maximum contact area of the foot paw remained ele-vated,and there was a statistically significant difference(P<0.05)between the combined administration group and PB21 and Dex alone.GAP43 and CGRP expression levels in synovial tissue were detected.The results indica-ted that PB21 and Dex alone could lower protein expression levels at 4 and 24 h at the two time points,and that the PB21+Dex group could still significantly lower GAP43 and CGRP expression levels at 48 h.At the 48 h time point,the PB21+Dex group was statistically significant when compared to the PB21 and Dex alone administration group(P<0.05).Conclusion In summary low dose dexamethasone can prolong the analgesic effect of PB21 on KOA rats,which is connected to reducing the expression of pain related proteins CGRP and GAP43.
		                        		
		                        		
		                        		
		                        	
3.Correlation between anxiety and perceived stress among medical personnel in a tertiary specialized cancer hospital
ZHUANG Qunfei ; WANG Jinyu ; LI Geling ; JIANG Shunjie ; SHEN Zhuping ; LI Qian
Journal of Preventive Medicine 2023;35(5):388-391
		                        		
		                        			Objective:
		                        			To examine the correlation between anxiety and perceived stress among medical personnel in a tertiary specialized cancer hospital, so as to provide the evidence for improving psychological health among medical personnel. 
		                        		
		                        			Methods:
		                        			In-service doctors, nurses and technicians were sampled from a tertiary specialized cancer hospital, and their demographics, perceived stress and sources of stress were collected using self-designed questionnaires. Anxiety was evaluated using the Self-rating Anxiety Scale (SAS), and the associations of anxiety with perceived stress and source of stress were examined using a multivariable logistic regression model. 
		                        		
		                        			Results:
		                        			A total 800 questionnaires were allocated and 655 valid questionnaires were recovered, with an effective recovery rate of 81.88%. The respondents included 160 men (24.43%) and 495 women (75.57%), and 297 respondents (45.34%) were at ages of 30 to 39 years. There were 14 respondents with no or low level of stress (2.14%), 245 with general level of stress (37.40%), 289 with high level of stress (44.12%), and 107 with extremely high level of stress (16.34%). The prevalence of anxiety was 36.64% among respondents. Multivariable logistic regression analysis identified perceived stress at work (OR=2.205, 95%CI: 1.657-2.675), doctor-patient relationship (OR=2.259, 95%CI: 1.561-3.282) and interpersonal relationship (OR=2.272, 95%CI: 1.387-3.693) as a factor affecting anxiety among medical personnel. 
		                        		
		                        			Conclusion
		                        			The anxiety correlates with stress at work, doctor-patient relationship and interpersonal relationship among medical personnel in a tertiary specialized cancer hospital.
		                        		
		                        		
		                        		
		                        	
4.Inhibition of ADAMTS by iPSC⁃MSCs in vitro protects cartilage matrix in patients with osteoarthritis
Feng Zhang ; Gang Cheng ; Yujiao Wu ; Zhuping Chu ; Xulei Wang ; Huimin Wang ; Kang Wang ; Wei Wei ; Shangxue Yan
Acta Universitatis Medicinalis Anhui 2023;58(9):1522-1528
		                        		
		                        			Objective  :
		                        			 To study the protective effect and mechanism of iPSC⁃MSCs on cartilage matrix in knee oste⁃
oarthritis (KOA) patients in vitro. 
		                        		
		                        			Methods  :
		                        			Cartilage tissues removed from KOA patients with joint replacement surgery were collected and subjected to tissue and cellular experiments , respectively.  Cartilage tissue was cut into small pieces and randomly divided into a control group , an IL⁃1β ( 10 ng/ml) induction group , and iPSC⁃MSCs 96 h and then co⁃cultured with different amounts of iPSC⁃MSCs ( 1 × 104 , 1 × 105 , 1 × 106 ) cells for 72 h.  For in tissues , the pathological changes of isolated cartilage tissues were examined by HE staining.  The levels of ADAMTS⁃4 , ADAMTS⁃5 , and type II collagen expression were analyzed by immunohistochemistry , while the levels of MMP13 , IL⁃6 , and IL⁃10 in culture supernatants were detected by ELISA kits. The 2 to 5 generations of chondrocytes , which were extracted from cartilage tissue of KOA patients , were stimulated with IL⁃1β ( 10 ng/ml) for 48 h and then co⁃cultured with different concentrations of iPSC⁃MSCs ( 1 × 104 , 1 × 105 , 1 × 106 ) cells for 72 h. Immunofluorescence and Western blot detected the expression of RUNX2 , ADAMTS⁃4 , and ADAMTS⁃5 in chondrocytes. 
		                        		
		                        			Results  :
		                        			Comparison with the control group , in the IL⁃1β⁃induced group , the levels of RUNX2 , ADAMTS⁃4 , and ADAMTS⁃5 increased , the level of type II collagen decreased , the levels of MMP⁃13 and IL⁃6 in the culture supernatant increased ( P < 0. 05) , and the level of IL⁃10 decreased ( P < 0. 05) ; Compared with the IL⁃1β⁃induced decreased the expression of RUNX2 , ADAMTS⁃4 , and ADAMTS⁃5 , promoted type II collagen expression and elevated IL⁃10 levels. 
		                        		
		                        			Conclusion  
		                        			iPSC⁃MSCs inhibited ADAMTS⁃4 and ADAMTS⁃5 expression in vitro , reduced cartilage extracellular matrix degradation , and played a role in articular cartilage protection.
		                        		
		                        		
		                        		
		                        	
5.Clinical characteristics and risk factors of acute mechanical small bowel obstruction in adults
Jing TIAN ; Feng WANG ; Zhuping ZHOU ; Min FENG ; Hao WANG ; Wenxian GUAN ; Jian HE ; Jun WANG
Chinese Journal of General Practitioners 2021;20(8):873-880
		                        		
		                        			
		                        			Objective:To analyze the clinical characteristics and the risk factors of acute mechanical small bowel obstruction in adults.Methods:The clinical data of 487 adult patients with acute mechanical small bowel obstruction treated in Drum Tower Hospital Clinical College of Nanjing Medical University from June 2010 to December 2020 were retrospectively analyzed. There were 259 cases of strangulated obstruction (strangulation group) and 228 cases of non-strangulated obstruction (simple group). The cases in the strangulation group were confirmed by operation or pathology, the cases in the simple group were confirmed by non-operative therapy ( n=167) or operation ( n=61). The clinical data, including age, abdominal pain, vomiting, tenderness, rebound pain and muscle defense; the CT signs, including transitional zone, dilatation of intestinal loop, high density of intestinal wall; laboratory test results, including white blood cell count, lactate dehydrogenase (LDH) and C-reactive protein (CRP) were analyzed in both groups. Univariate and multivariate regression were used to identify risk factors of strangulated small bowel obstruction. Results:There were significant significances in gender (male vs.female: χ2=4.35, OR=0.67), rebound pain (χ2=170.98, OR=45.12), muscle tension(χ2=113.79, OR=29.32), American Society of Anesthesiologists score (ASA score)≥3 (χ2=12.94, OR=51.58), white blood cell count ( t=6.19, OR=1.14), LDH ( t=2.37, OR=1.00), CRP ( t=2.23, OR=1.01), albumin ( t=2.04, OR=0.97), mesenteric fluid sign (χ2=49.44, OR=5.40), increased bowel wall density (χ2=222.78, OR=62.66), bowel wall thickening sign (χ2=43.81, OR=3.49), ascites (χ2=237.29, OR=43.01), beak sign (χ2=231.50, OR=56.62), mesenteric fat stranding (χ2=242.65, OR=34.90), and stranding sign(χ2=224.79, OR=53.48) between strangulation group and simple group ( P<0.01). The multivariate regression analysis showed that mesenteric fluid sign ( OR=12.94), muscle tension ( OR=7.28), ascites ( OR=6.42), increased bowel wall density ( OR=4.30), bowel wall thickening sign ( OR=1.85), white blood cell count ( OR=1.14) and gender (male vs. female: OR=0.50) were risk factors of strangulated small bowel obstruction. Conclusion:In acute mechanical small bowel obstruction,for female patients presenting mesenteric fluid sign, muscle tension, ascites, increased bowel wall density, bowel wall thickening sign and increased white blood cell count, the strangulated obstruction is likely to occur.
		                        		
		                        		
		                        		
		                        	
6.Prokaryotic expression, purification and functional identification of epidermal pattern factors in Arabidopsis thaliana.
Zhuping JIN ; Cheng LI ; Lei WANG ; Yanxi PEI
Chinese Journal of Biotechnology 2020;36(4):792-800
		                        		
		                        			
		                        			Stomatal density is important for crop yield. In this paper, we studied the epidermal pattern factors (EPFs) related to stomatal development. Prokaryotic expression vectors were constructed to obtain EPFs. Then the relationship between EPFs and hydrogen sulfide (H2S) was established. First, AtEPF1, AtEPF2 and AtEPFL9 were cloned and constructed to pET28a vectors. Then recombinant plasmids pET28a-AtEPF1, pET28a-AtEPF2 and pET28a-AtEPFL9 were digested and sequenced, showing successful construction. Finally, they were transformed into E. coli BL21(DE3) separately and induced to express by isopropyl β-D-galactoside (IPTG). The optimized expression conditions including IPTG concentration (0.5, 0.3 and 0.05 mmol/L), temperature (28 °C, 28 °C and 16 °C) and induction time (16 h, 16 h and 20 h) were obtained. The bands of purified proteins were about 18 kDa, 19 kDa and 14.5 kDa, respectively. In order to identify their function, the purified AtEPF2 and AtEPFL9 were presented to Arabidopsis thaliana seedlings. Interestingly, the H2S production rate decreased or increased compared with the control, showing significant differences. That is, EPFs affected the production of endogenous H2S in plants. These results provide a foundation for further study of the relationship between H2S and EPFs on stomatal development, but also a possible way to increase the yield or enhance the stress resistance.
		                        		
		                        		
		                        		
		                        			Arabidopsis
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		                        			genetics
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		                        			metabolism
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		                        			Arabidopsis Proteins
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		                        			genetics
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		                        			isolation & purification
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		                        			metabolism
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		                        			Escherichia coli
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		                        			genetics
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		                        			Genetic Vectors
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		                        			genetics
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		                        			Hydrogen Sulfide
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		                        			metabolism
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		                        			Plasmids
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		                        			genetics
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		                        			Seedlings
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		                        			metabolism
		                        			
		                        		
		                        	
7.Research progress on sensitive indexes of nursing quality of patients with cancer pain
Chinese Journal of Modern Nursing 2020;26(29):4131-4135
		                        		
		                        			
		                        			This paper reviews the framework and content of sensitive indexes of nursing quality of patients with cancer pain. Based on Donabedian's quality management theory, it elaborates from three dimensions of structure, process and result, aiming to provide a reference for the construction of unified and standardized sensitive indexes of nursing quality for patients with cancer pain.
		                        		
		                        		
		                        		
		                        	
8.Expression of ubiquitin editing enzyme A20 and its pathway in steatotic hepatocytes and monocytes
Luoyan AI ; Qingqing XU ; Changwei WU ; Dazhi SU ; Xiaohan WANG ; Zhiwei CHEN ; Zhuping FAN
Chinese Journal of Digestion 2015;35(4):247-251
		                        		
		                        			
		                        			Objective To investigate the changes of A20 expression stimulated by free fatty acids (FFA) and its pathway.Methods HepG2 cells and U937 cells were stimulated by 0.5 mmol/L mixed FFA.The expression of A20,phosphor-p65 and phosphor-IκBα of neclear factor (NF)-κB pathway and phosphor-c-Jun N-terminal kinase (JNK),JNK,phosphor-extracellular signal-regulated kinase (ERK),ERK,phosphor-p38 and p38 of mitogen-activated protein kinase (MAPK) pathway were detected by Western blotting.The level of interleukin (IL)-12p,IL-1β,tumor necrosis factor (TNF)-α,IL-6,IL-10 and IL-8 cytokines in the supernatant of cell culture were detected by flow cytometry.T-test was performed for statistical analysis.Results The level of A20 changed along with the stimulated time of FFA.NF-κB and MAPK pathways were activated after FFA stimulation.The secretion of IL-6 and IL-8 increased after HepG2 cells stimulated by FFA and both reached peak at 24 hour.Compared with control group,the difference in IL-8 was statistically significant ((423.8 ± 8.9) pg/mL vs (12.4 ± 4.5) pg/mL,t=41.28,P<0.01).The difference in IL-6 was also statistically significant ((4 082±423.6) pg/mL vs (52.9±29.5) pg/mL,t=9.49,P<0.01).After U937 cells were stimulated by FFA,the secretion of IL-8 increased compared with control group.And in a certain period of time the secretion was time dependence.The maximum secretion of 24 hours was (200.6±5.7) pg/mL vs (5.0±3.9) pg/mL,and the difference was statistically significant (t=28.16,P<0.01).IL-10,IL-12p,IL-1β and TNF-α were detected.Both NF-κB pathway and MAPK pathway were detected.Conclusions The in vitro FFA mediated steatotic cell model could induce the expression change of A20 and secretion of inflammatory cytokines.NF-κB and MAPK pathways involved in the response to FFA in HepG2 cells and U937 cells.
		                        		
		                        		
		                        		
		                        	
9.Immunophenotypic analysis of abnormal plasma cell clones in bone marrow of primary systemic light chain amyloidosis patients.
Yang HU ; Mangju WANG ; Yan CHEN ; Xue CHEN ; Fang FANG ; Shiqin LIU ; Ying ZHANG ; Xueqiang WU ; Ping ZHU
Chinese Medical Journal 2014;127(15):2765-2770
BACKGROUNDPrimary systemic light chain amyloidosis (AL) is a rare plasma cell disease, our purpose was to analyze the immunophenotypic characteristics of the plasma cells in bone marrow in AL patients, and explore whether the detection of abnormal plasma cell clones in bone marrow by flow cytometry (FCM) could be used as an important indicator of AL diagnosis.
METHODSFresh bone marrow samples were collected from 51 AL, 21 multiple myeloma (MM), and 5 Waldenström's macroglobulinemia (WM) patients. The immunophenotype of bone marrow cells were analyzed and compared by FCM using a panel of antibodies including CD45, CD38, CD138, CD117, CD56, and CD19.
RESULTSIn AL, light chain restriction could be identified in 31 cases (60.9%), in which the λ light chain restriction was found in 24 cases (77.4%). In MM, κ light chain restriction was found in 13 cases (61.9%), and λ light chain restriction in eight cases. CD45 on abnormal plasma cells was negative to weakly positive in both AL and MM, but was positive to strongly positive in WM. In the bone marrow plasma cells of the 51 AL, 78.4% were CD56+, 68.6% were CD117+, and 88.2% were CD19-. While in the 21 MM cases, 66.7% were CD56+, 38.1% were CD117+, and 90.4% were CD19-. The plasmacytoid lymphocytes in the five WM patients were CD19+ and CD56-, CD117-.
CONCLUSIONDetection of abnormal plasma cell clones in bone marrow by FCM is valuable for the diagnosis of AL.
Adult ; Aged ; Aged, 80 and over ; Amyloidosis ; immunology ; metabolism ; CD56 Antigen ; metabolism ; Female ; Flow Cytometry ; Humans ; Immunoglobulin Light-chain Amyloidosis ; Immunoglobulin lambda-Chains ; metabolism ; Immunophenotyping ; Leukocyte Common Antigens ; metabolism ; Male ; Middle Aged ; Multiple Myeloma ; immunology ; metabolism ; Proto-Oncogene Proteins c-kit ; metabolism ; Waldenstrom Macroglobulinemia ; immunology ; metabolism
10.Effect of serum IGF-1, IGFBP-3 and HtSDS in rhGH-treated SGA children
Haiying CHENG ; Sheng LIU ; Zhuping WANG ; Yandan YIN ; Xue FENG ; Jin CHEN
Chinese Journal of Primary Medicine and Pharmacy 2014;21(12):1816-1819
		                        		
		                        			
		                        			Objective To investigate the clinical value of serum insulin-like growth factor-1 (IGF-1),insulin-like growth factor-binding protein-3 (IGFBP-3) and mean height standard deviation score(HtSDS) in recombinant human growth hormone (rhGH)-treated small for gestational age (SGA) children.Methods 32 cases of SGA failed of catch-up growth were subcutaneously injected with rhGH every night before bed at a dose of 0.15-0.20u· kg-1 · d-1.And the variation of serum IGF-1,IGFBP-3,HtSDS,body height and growth velocity (GV) were monitored at 3,6,9 and 12 months of the treatment and compared.Results The GV (cm/years) increased to (12.4 ± 3.2),(11.0 ± 2.3),(10.1 ± 3.5),(9.4 ± 1.8) versus (4.1 ± 0.5) before treatment,suggesting a significant catch-up growth after treatment (F =51.35,P < 0.01).Accordingly,HtSDS increased to (-2.55 ± 0.73),(-2.39 ± 0.65),(-2.21 ± 0.58),(-2.09 ± 0.94) versus (-2.81 ± 0.64),suggesting that height gaps were reduced compared to the normal children of the same age and sex after rhGH treatment (F =4.99,P <0.01).The serum levels of IGF-1,IGFBP-3 were significantly increased compared to pre-treatment (F =34.52,14.04,all P < 0.01),especially for serum IGF-1,which maintained at a high level after 3 months of treatment and up to peak after 6 months.However,the ratio of IGF-1/IGFBP-3 showed no increase after treatment (F =1.82,P > 0.05).There was a significant positive correlation between the serum level of IGF-1 and △HtSDS after the rhGH treatment within six months (r3 =0.72,r6 =0.91),while there was no correlation between them after 9 months (r9 =0.26,r12 =0.33).There were no serious adverse events during the rhGH treatment.Conelusion The serum levels of IGF-1,IGFBP-3 were significantly increased after the rhGH treatment,especially for serum IGF-1.There was a significant positive correlation between the serum level of IGF-1 and △HtSDS within six months,but not after 9 months.The rhGH treatment could be a safe and effective strategy for SGA children.
		                        		
		                        		
		                        		
		                        	
            

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