1.Effects of Gubi Decoction on serum related inflammatory factors and PI3K/Akt/mTOR signaling pathway in osteoarthritis model rats
Liqing ZHONG ; Shaobo DING ; Runkai HU ; Weichao HAN ; Shufen HE ; Baocheng XIE
International Journal of Traditional Chinese Medicine 2021;43(4):357-362
Objective:To explore the effect of Gubi Decoction on serum related inflammatory factors and PI3K/Akt/mTOR signaling pathway in osteoarthritis model rats. Methods:Seventy SPF rats were randomly divided into the blank group, sham operation group, Glucosamine sulfate group, and the low, medium and high dose Gubi Decoction groups. Except the blank group and sham operation group, knee osteoarthritis animal models were prepared by the modified Hulth method in each group. On the 28th day after successful model preparation, the high, medium and low dose Gubi Decoction groups were given Gubi Decoction 24, 12 and 6 g/kg by gavage respectively; glucosamine sulfate group was given glucosamine sulfate tablet suspension 3 g/L by gavage, once a day for 28 days. The levels of TNF-α and IL-1β in serum were detected by ELISA. The gene expressions of PI3K, Akt and mTOR in cartilage tissue were detected by Real-PCR. The protein expressions of PI3K, Akt, p-PI3K, p-Akt and mTOR were detected by Western blot. Results:Compared with the model group, the knee joint diameter[(11.17 ± 1.81) mm, (11.60 ± 1.38) mm, (10.80 ± 1.17) mm vs. (12.57 ± 0.98) mm] of the rats in the glucosamine sulfate group and the medium and high dose Gubi Decoction groups significantly decreased ( P<0.05). The content of TNF-α [(111.43 ± 21.98) ng/L, (53.42 ± 13.25) ng/L vs. (157.89 ± 23.60) ng/L], IL-1β [(67.50 ± 18.44) ng/L, (48.22 ± 9.63) ng/L vs. (96.11 ± 14.85) ng/L] in the medium and high dose Gubi Decoction groups significantly decreased ( P<0.05), and the expression of PI3K (1.87 ± 0.17, 1.24 ± 0.49 vs. 2.19 ± 0.47), Akt (1.50 ± 0.51, 1.10 ± 0.32 vs. 2.68 ± 0.63), and mTOR (1.32 ± 0.54, 1.10 ± 0.33 vs. 2.94 ± 0.55) mRNA in the medium and high dose Gubi Decoction groups significantly decreased ( P<0.05). The expression of PI3K, Akt, p-PI3K, p-Akt in the low, medium and high dose Gubi Decoction groups significantly decreased ( P<0.05), and the expression of mTOR in the medium dose Gubi Decoction group significantly decreased ( P<0.05). Conclusion:Gubi Decoction can significantly reduce the level of inflammatory factors in the serum of osteoarthritis model rats, and its anti-inflammatory and analgesic effects may be related to PI3K/Akt/mTOR signaling pathway.
2.A case of vertical one-and-a-half syndrome caused by unilateral thalamo-mesencephalic infarct
Shilin YANG ; Shufen CHEN ; Shenyi KUANG ; Zhiyuan DONG ; Huiying WANG ; Xiang HAN
Chinese Journal of Neurology 2021;54(6):593-596
Vertical one-and-a-half syndrome (VOHS) is a rare ophthalmoplegia syndrome caused by unilateral thalamo-mesencephalic infarct. It is manifested as a conjugate upgaze palsy and a monocular paresis of downward gaze. Recognition of VOHS helps clinical localization. No cases of VOHS with photos or videos of ocular motility have been reported in China. Herein a case of VOHS caused by unilateral thalamo-mesencephalic infarct is reported, in order to improve awareness about the ocular sign.
4.Effects of leucine on adipogenesis in 3T3-L1 preadipocytes during and after differentiation
Jiangtao YANG ; Jiaying XU ; Jun JIAO ; Ru ZHANG ; Shufen HAN ; Liqiang QIN
Chinese Journal of Preventive Medicine 2016;50(6):535-540
Objective To observe the effects of leucine on adipogenesis in 3T3-L1 preadipocyte during and after differentiation, and to investigate possible mechanisms. Methods Respectively, 0.0 (control), 0.5, 1.0 and 2.0 mmol/L leucine was added in 3T3-L1 cells and cell proliferation was measured by MTT. Then, 3T3-L1 preadipocyte was induced to differentiate. Leucine was added during whole differentiation period, or after differentiation for 4 days. The cells were stained with Oil Red O dye to observe lipid droplet. The culture media were collected and used to determine glycerol contents. Meanwhile, protein expressions related to lypolytic enzymes, leptin signaling pathway were determined by Western blot. Results MTT result showed that cell viabilities were (100.00 ± 12.10)%, (102.73 ± 12.38)%, (103.94 ± 14.65)%, (108.70 ± 5.05)% in 0.0, 0.5, 1.0 and 2.0 mmol/L leucine groups, respectively, there were no significant differences in cell proliferation among 4 groups (F=1.07, P=0.383). When 0.0, 0.5, 1.0 and 2.0 mmol/L leucine was added during differentiation, the relative number of lipid droplet was 1.00 ± 0.06, 0.94±0.09, 0.82±0.08 and 0.79±0.04, respectively (F=11.74, P<0.001), and it was significantly lower in 1.0 and 2.0 mmol/L leucine groups than in control group(P=0.002 and P<0.001, respectively). There was no significant difference in lipid droplet when leucine was added after differentiation (F=0.16, P=0.924). When leucine was added during differentiation, the increment of glyceride contents in medium was (65.04 ± 11.75) , (71.45 ± 23.71), (79.37 ± 17.63) and (110.32 ± 25.36) μmol/L, respectively (F=2.92, P=0.100) . And it was significantly higher in 2.0 mmol/L leucine group (110.32 ± 25.36)μmol/L than in control group (65.04 ± 11.75)μmol/L(t=2.73, P=0.026). No significant difference of the increment of glyceride contents among 4 groups was observed when leucine was added after differentiation (F=0.80, P=0.528). Western blot results showed that leucine treatment during differentiation upregulated expression level of hormone-sensitive lipase phosphorylation (after 0.0 and 2.0 mmol/L leucine treatment,the protein levels were 1.00 ± 0.08 vs. 2.54 ± 0.27, P<0.001), and downregulated the protein expression levels of perilipin A, leptin and leptin-related pathway, such as leptin receptor, Janus kinase 2 and suppressor of cytokine signaling-3 (after 0.0 and 2.0 mmol/L leucine was added, the protein levels were(1.00 ± 0.03)vs.(0.31 ± 0.07),(1.00 ± 0.08)vs.(0.22±0.07),(1.00±0.07)vs.(0.21 ± 0.04),(1.00 ± 0.03)vs.(0.35 ± 0.05),(1.00 ± 0.06)vs.(0.34 ± 0.05), P<0.001). Leucine treatment after differentiation had no effects on these protein expressions (all P>0.05). Conclusion Leucine inhibits adipogenesis during 3T3-L1 preadipocyte differentiation by the regulation of lypolytic enzymes and leptin signaling pathway;however, leucine has no effect on adipogenesis when differentiation completed.
5.Effects of leucine on adipogenesis in 3T3-L1 preadipocytes during and after differentiation
Jiangtao YANG ; Jiaying XU ; Jun JIAO ; Ru ZHANG ; Shufen HAN ; Liqiang QIN
Chinese Journal of Preventive Medicine 2016;50(6):535-540
Objective To observe the effects of leucine on adipogenesis in 3T3-L1 preadipocyte during and after differentiation, and to investigate possible mechanisms. Methods Respectively, 0.0 (control), 0.5, 1.0 and 2.0 mmol/L leucine was added in 3T3-L1 cells and cell proliferation was measured by MTT. Then, 3T3-L1 preadipocyte was induced to differentiate. Leucine was added during whole differentiation period, or after differentiation for 4 days. The cells were stained with Oil Red O dye to observe lipid droplet. The culture media were collected and used to determine glycerol contents. Meanwhile, protein expressions related to lypolytic enzymes, leptin signaling pathway were determined by Western blot. Results MTT result showed that cell viabilities were (100.00 ± 12.10)%, (102.73 ± 12.38)%, (103.94 ± 14.65)%, (108.70 ± 5.05)% in 0.0, 0.5, 1.0 and 2.0 mmol/L leucine groups, respectively, there were no significant differences in cell proliferation among 4 groups (F=1.07, P=0.383). When 0.0, 0.5, 1.0 and 2.0 mmol/L leucine was added during differentiation, the relative number of lipid droplet was 1.00 ± 0.06, 0.94±0.09, 0.82±0.08 and 0.79±0.04, respectively (F=11.74, P<0.001), and it was significantly lower in 1.0 and 2.0 mmol/L leucine groups than in control group(P=0.002 and P<0.001, respectively). There was no significant difference in lipid droplet when leucine was added after differentiation (F=0.16, P=0.924). When leucine was added during differentiation, the increment of glyceride contents in medium was (65.04 ± 11.75) , (71.45 ± 23.71), (79.37 ± 17.63) and (110.32 ± 25.36) μmol/L, respectively (F=2.92, P=0.100) . And it was significantly higher in 2.0 mmol/L leucine group (110.32 ± 25.36)μmol/L than in control group (65.04 ± 11.75)μmol/L(t=2.73, P=0.026). No significant difference of the increment of glyceride contents among 4 groups was observed when leucine was added after differentiation (F=0.80, P=0.528). Western blot results showed that leucine treatment during differentiation upregulated expression level of hormone-sensitive lipase phosphorylation (after 0.0 and 2.0 mmol/L leucine treatment,the protein levels were 1.00 ± 0.08 vs. 2.54 ± 0.27, P<0.001), and downregulated the protein expression levels of perilipin A, leptin and leptin-related pathway, such as leptin receptor, Janus kinase 2 and suppressor of cytokine signaling-3 (after 0.0 and 2.0 mmol/L leucine was added, the protein levels were(1.00 ± 0.03)vs.(0.31 ± 0.07),(1.00 ± 0.08)vs.(0.22±0.07),(1.00±0.07)vs.(0.21 ± 0.04),(1.00 ± 0.03)vs.(0.35 ± 0.05),(1.00 ± 0.06)vs.(0.34 ± 0.05), P<0.001). Leucine treatment after differentiation had no effects on these protein expressions (all P>0.05). Conclusion Leucine inhibits adipogenesis during 3T3-L1 preadipocyte differentiation by the regulation of lypolytic enzymes and leptin signaling pathway;however, leucine has no effect on adipogenesis when differentiation completed.
6.The application of acridine orange fluorescent staining method in circulating tumor cell
Min LIU ; Fuling MA ; Rong HAN ; Mei LI ; Shufen LIU ; Shumin ZHANG
Tianjin Medical Journal 2015;(7):792-795
Objective To establish the screening platform of circulating tumor cells (CTCs) using acridine orange fluo?rescent (AO-F) dyeing method, and to apply it in the screening of peripheral blood CTCs in patients with kidney cancer. Methods Twenty-seven patients with metastatic renal cell carcinoma was included in this study. Primitive tumor cells and kidney cancer cell line 769-P were cultured with different concentrations of fetal bovine serum. Smears were prepared and observed under fluorescence microscopy. The percentage of AO-F positive staining of 769-P cells under 5 random sights was calculated. The sensitivity of AO-F staining to cells was evaluated. The 5 mL morning fasting venous blood was obtained from 10 subjects with healthy check-up. The 1×106 cell suspension was prepared. The logarithmic phase of renal tumor cells was used to prepare tube containing 500, 200, 100, 50 and 10 tumor cell suspension, which were mixed with 1×106 nucleated cells to establish CTCs model of renal cancer. AO-F staining method was used to detect the expression of AO-F positive cells. The correlation between expression of AO-F positive cells and clinical parameters was analyzed. Results The prima?ry cells and cell line 769-P showed similar bright color and morphological characteristics. The percentage of AO-F positive staining in 769-P cells was 93%±3%under 5 random sights. The recovery rates (%) of four groups (500, 200, 100 and 50 tu?mor cell suspension) were 10.2±3.8, 9.2±2.3, 10.8±2.6 and 10.5±1.9, respectively. There were no significant differences in recovery rates between four groups (P>0.05). The group of 10 tumor cell suspension could find AO-F positive staining cells occasionally. Zero case was positive in controls. Nine of 27 patients were positive and the rate was 33.33%. There were no significant statistical differences in AO-F positive rates between gender, age, tumor size, pathological pattern, Furhman stage, metastasis of lung and presence of tumor (P>0.05). Conclusion It is confirmed that the method of CTCs staining with AO-F, which has high specificity and reproducibility, is feasible to detect CTCs and worthy of being studied. There is a certain reference value to predict tumor recurrence and metastasis.
7.Effects of compound whole grain-soybean on insulin resistance and serum adipocytokines in impared fasting glucose population.
Shufen HAN ; Hong ZHANG ; Jing CHI ; Yaqi LIU ; Siyu ZHOU ; Chengkai ZHAI
Chinese Journal of Preventive Medicine 2014;48(1):23-27
OBJECTIVETo evaluate the effects of compound whole grain-soybean on insulin resistance and serum adipocytokines levels in impared fasting glucose population.
METHODSAccording to inclusion and exclusion criteria, 163 cases of impared fasting glucose (IFG) Chinese Han population from the age of 40 to 75 years old, were screened from 12 community health centers of three main districts of Nanjing city by the multi-stage cluster and simple randomization method from March to September, 2008. The IFG subjects were randomly divided into the intervention group (87 individuals) and control group (76 individuals) by quasi-experimental design. The intervention group was provided with compound whole grain-soybean and health education, while only health education was provided for the control group. Body mass index (BMI), waist-to-hip ratio (WHR), lipid profiles, fasting blood glucose (FBG), fasting insulin (FINS) and homeostasis model assessment of insulin resistance (HOMA-IR), adipocytokines including leptin, lipocalin 2 (LCN-2) and adiponectin (ADP) levels were measured before and after the half a year intervention period. Chi square test was used to analyze the distribution differences. Two-sample t-test was used to compare the differences of the two groups before and after the half a year intervention period, and paired t-test was used to compare the differences between before and after intervention in the intervention group or control group. Wilcoxon rank sum test was used to compare the differences of all indexes between after and before dietary intervention.
RESULTSAfter dietary intervention for half a year on the IFG population, BMI ((24.87 ± 3.69) kg/m(2)), FBG((6.27 ± 0.24) mmol/L), FINS((7.14 ± 1.05) mU/L) , HOMA-IR (1.99 ± 0.31), leptin ((13.07 ± 2.22) µg /L), LCN-2 ((67.42 ± 18.20) µg/L) of intervention group were decreased significantly compared to the levels of BMI ((25.16 ± 4.07) kg/m(2)), FBG((6.40 ± 0.28) mmol/L), FINS ((7.32 ± 1.54) mU/L), HOMA-IR (2.08 ± 0.45), leptin ((13.43 ± 2.52) µg/L), LCN-2((74.87 ± 17.81) µg/L) before dietary intervention, t values were 4.48, 7.08, 2.05, 3.39, 3.28 and 6.36, respectively, and all P values were < 0.05, while ADP ((5.07 ± 1.51) mg/L) of intervention group after dietary intervention was increased significantly compared to the level of ADP ((4.92 ± 1.53) mg/L) before dietary intervention, t = -2.47 and P < 0.05. The medians (P25, P 75) of differences after and before dietary intervention in the intervention group were BMI (-0.25(-0.68, 0.02) kg/m(2)), FBG (-0.08 (-0.20, 0.00) mmol/L), FINS (-0.15(-0.32, 0.00) mU/L), HOMA-IR (-0.07(-0.12, -0.03)), leptin (-0.36(-0.77, 0.12) µg/L), LCN-2 (-5.85(-14.29, -0.71) µg/L) and ADP (0.15(-0.13, 0.36) mg/L), and the medians of differences of after and before dietary intervention in the control group were BMI (0.00(-0.23, 0.29) kg/m(2)), FBG (0.00(-0.03, 0.04) mmol/L), FINS (-0.01(-0.13, 0.04) mU/L), HOMA-IR (-0.01(-0.05, 0.02)), leptin (-0.07 (-0.57, 0.46) µg/L), LCN-2 (-0.85(-5.39, 1.63) µg/L) and ADP (0.02(-0.19, 0.13) mg/L). There were significantly statistical differences between them (Z values were -3.65, -4.88, -3.08, -5.23, -2.16, -4.43 and 3.05, all P values were <0.05).
CONCLUSIONDietary intervention of compound whole grain-soybean can improves glucose level, increase insulin sensitivity and ameliorate insulin resistance state of IFG population.
Adipokines ; blood ; Adiponectin ; blood ; Aged ; Apolipoproteins ; blood ; Blood Glucose ; metabolism ; Female ; Humans ; Insulin ; blood ; Insulin Resistance ; Male ; Middle Aged ; Soybeans ; Waist-Hip Ratio
8.Analysis on the Results of Blood Lead of Children in Panyu,Guangzhou
Xing XIAO ; Jinhua HE ; Yuguang LI ; Zeping HAN ; Shufen HU ; Hui HUANG
Journal of Modern Laboratory Medicine 2014;(6):145-147
Objective To understand the children health care and the prevention of lead pollution in Panyu in the future by re-search on the detection result of blood lead from Panyu children during 2011 and 2012.Methods The detection of whole blood lead was by atomic absorption spectrometer from body examination in 6 308 children in Panyu Center Hospital be-tween 2011 and 2012:①The blood lead result from assigned children were divided into five age groups:infancy group (1~12 months,1 511 cases),toddler age group (1~2 years,1 558 cases),preschool age group (3~6 years,1 704 cases),school age group (7~11 years,1 076 cases)and adolescence group (12~18 years,459 cases).②Based on different age group,each group was separated by male and female.③According to the detection year,each group was also put into 2011 and 2012 re-spectively.Results The overall level of children blood lead in Panyu,Guangzhou in 2011 and 2012 was 37.01±29.18μg/L, and the total poisoning rate was 1.2 9%.Along with the growth of age,the overall level of children blood lead was in up-wards trend but the poisoning rate kept in a stable level beginning from infant stage.The blood lead level of boys in Panyu was 38.87±30.18μg/L,and the poisoning rate was 1.91%;the blood lead level of girls was 34.13±27.32μg/L,and the poisoning rate was 1.29%.The blood lead levels and the poisoning rates of boys were higher than girl’s in most of age groups.Conclusion Compared with other big cities like Guangzhou,the blood lead level of children in Panyu was declined but it’s not the standard performance,and there’s a certain extent of lead pollution.So in addition to reinforce pollution con-trol from government,parents and children should enhance the knowledge,education and precaution of lead poisoning.
9.The literature research on current residency standardized training in China
Qichao NIU ; Shufen YANG ; Bing HAN ; Yang YANG
Chinese Journal of Medical Education Research 2014;13(12):1267-1271
Objective To sum up the status and existing problems of standardized training of resident physician in China,and to promote residency standardized training.Methods With essayas retrieving items,resident + standardization training as the search term,all relevant papers published in CNKI from January 2004 to January 2014 with a title containing residency standardized training were reviewed.By means of literature analysis,the literature in 8 aspects was classified including the year of publication,status of journal,training base selection conditions,training objects and training time,content,specialist fields,management,assessment methods and shortages.Excel was used to make descriptive analysis of the results.Results After elimination,we took 152 papers as experimental group.In detail,12 literatures about training base selection conditions demonstrated that the level and type of hospital was the main criteria for training base;28 literatures about training objects and training time indicated that trainees were oriented to the pattern of 5+3,the latitude of PhD and Masters could be decreased properly; 36 literatures about training content focused on clinical skills and expertise.Only 19 literatures was concerned with specialist fields,which needed enriching.12 articles about management implemented management of supervisor responsibility system.22 literatures about the assessment method recommend that priority should be given to clinical skills examination to explore the hospital problems of resident standardization training.Conclusion The standardized training of resident physician has been carried out in China,but there are still some problems which need to be further improved.
10.Investigation of the awareness of the oral care with machinery ventilates among new nurses
Shuyan MA ; Zhongping YIN ; Shufen HAN ; Ping LI ; Yun HUANG ; Xin YOU ; Yanhua XU
Chinese Journal of Modern Nursing 2014;20(6):644-646
Objective To investigate the awareness of the oral care with machinery ventilates among new nurses , and provide reference for nursing pre-service training plan .Methods Totals of 108 new nursing staff from comprehensive hospitals was recruited , investigating the awareness of the oral care in machinery ventilates with a self-designed questionnaire and analyzing the results .Results Nurses involved in professional oral care training showed no statistical difference between the different levels of education background (χ2 =0.183, 0.077, 0.000, respectively;P>0.05).The score of machinery ventilates importance in patients with sputum care was (7.9 ±3.0).The score of endotracheal intubation care was (7.6 ±2.8).The score of nursing record was (7.1 ±2.6), and score of basic oral care importance was (5.9 ±2.7).Conclusions New nurses have a low level of oral care knowledge for machinery ventilates , and more detailed oral care training plan should guide them in the subsequent stage of continuing education .

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