1.Primary astroeytes with mutant SOD1G93A are susceptible to oxidative stress
Weisong DUAN ; Hui BU ; Yansu GUO ; Zhongyao LI ; Chunyan LI
Basic & Clinical Medicine 2010;30(4):378-382
Objective To study the vulnerability of astrocytes bearing mutant SOD1 under the oxidative stress.Methods The cytotoxicity of the serum deprived astrocytes was measured by MTT.The level of ROS was shown by fluorescence of DCF through confocal microscopy.The expression of Nrf2,HO1 and NQO1 in the different cells was detected by Western blot.Results The level of cellular toxicity was higher in the astrocytes bearing mutant SOD1 exposed to the oxidative stress than the astrocytes hearing wild type SOD1.In the astrocytes bearing mutant SOD1,the expression of Nrf2,HO1 and NQO1 decreased.In the presence of mutant SOD1,an unexpected 44 per-cent decrease of Nrf2 was detected.This was associated with a decreases in multiple downstream phase Ⅱ detoxif-ying enzymes and antioxidant enzymes,known as NQO1 and HO1.Furthermore,our results showed that the ex-pression of NQO1 increased 1.5 and 2.5-fold by EGCG at 5 and 10 μmol/L.EGCG also elevated the expression of total Nrf2.Confocal microscopy showed that EGCG caused Nrf2 translocation from the cytoplasm to the nucleus.Conclusion Decrease in Nrf2 expression is the mechanism to explain the vulnerability of astrocytes bearing mutant SOD1 and EGCG strengthened antioxidation function by upregulating the activity of Nrf2.
2.A missense mutation outside the large pocket of the retinoblastoma protein
Huangxuan SHEN ; Qingjiong ZHANG ; Xueshan XIAO ; Shiqiang LI ; Xiangming GUO ; Zhongyao WU
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To understand the effect of the RB1 gene mutation on the function of pRB (the protein product of the RB1 gene) in the patients with retinoblastoma (RB). METHODS: The genomic DNA from retinoblastoma patients was extracted. After amplification, the promoter and all 27 exons were screened by SSCP-heteroduplex method. The mutation was cloned and identified by sequencing. The effect of the mutation product on the function of pRB was analyzed. RESULTS: One missense mutations of the exon 4 of the RB1 gene was identified in the genomic DNA from RB patients. This mutation was outside the large pocket of the pRB. No mutation of the RB1 gene was found in the genome DNA of the patient's parents. This is the fourth report that there was a genome mutation located outside the large pocket of pRB in the RB patients. CONCLUSION: The amino-terminus of the pRB may be essential for growth suppression.
3.Cloning and Expression of GL-7-ACA Acylase in E.coli
Hui LUO ; Xiaojia HU ; Hang ZHOU ; Yizhou TONG ; Hui YU ; Qiang LI ; Zhongyao SHEN ;
Microbiology 1992;0(04):-
To facilitate the expression of GL 7 ACA acylase gene in a recombinant E coli , a fragment of the gene, in which the signal peptide was deleted by PCR method, was inserted into a prokaryotic expression vector, pET 28a By colony PCR method screening, a recombinant plasmid pET ACY was obtained and then transformed into the expression host BL21 (DE3) The influences of induction conditions such as IPTG concentration, the time of induction and the induction temperature on the expression of the recombinant protein were investigated Under optimal condition, the enzyme activity could reach 266 U/L Finally, the recombinant GL 7 ACA acylase can be easily isolated to a purity of about 80% by a simple anion ion exchange chromatography with enzyme activity recovery of 50%
4.Study on effect of different extraction methods on anti-inflammatory and analgesic activity of Anemone hupehensis
Xiang LI ; Yihong XU ; Xiaohui SUN ; Zhongyao HAN ; Yunyan FU ; Junke WU
Chongqing Medicine 2018;47(15):1990-1992,1996
Objective To investigate the effect of different extraction methods on anti-inflammatory and analgesic activity of Anemone hupehensis.Methods The different abstracts were prepared from the whole herb of Anemone hupehensis.The analgesic effect was observed by adopting the mouse torsion and electric heating plate method,and the anti-inflammatory activity was comprehensively evaluated by using the mouse ear tumefaction,toe tumefaction and tampon granulation tumefaction exprements.Results Compared with the blank model group,the anti-inflammatory action difference of low dose in the water layer parts of mouse ear tumefaction,toe tumefaction and tampon granulation tumefaction had no statistical significance(P>0.05),and the extracting parts of rest doses all had significant anti-inflammatory and analgesic effect (P<0.05).Ethyl acetate part had strongest activity in the electric heating plate experiment.N-butanol part had strongest activity in the ear tumefaction,toe tumefaction,tampon granulation tumefaction experiments and torsion method.Conclusion The whole herb of Anemone hupehensis has prominent anti-inflammatory and analgesic effect,and the ethyl acetate part E and N-butanol part are main effective parts.
5.Short-term efficacy of hip arthroscopic surgery assisted by platelet-rich plasma in the treatment of femoroacetabular impingement syndrome
Zhongyao LI ; Mingyang AN ; Yidong WU ; Kangkang YU ; Boda WANG ; Yibo LI ; Dongqiang GU ; Yaoting WANG ; Long WANG ; Mingxin WANG ; Jiapeng ZHENG ; Chunbao LI
Chinese Journal of Trauma 2023;39(10):885-892
Objective:To compare the short-term efficacy of hip arthroscopic surgery assisted by platelet-rich plasma (PRP) and hip arthroscopy alone in the treatment of femoroacetabular impingement (FAI).Methods:A retrospective cohort study was performed on the clinical data of 133 FAI patients admitted to Fourth Medical Center of PLA General Hospital from January 2019 to January 2021. The patients included 86 males and 47 females, aged 19-71 years [(39.1±12.6)years]. A total of 67 patients were treated with hip arthroscopy alone (hip arthroscopy group), and 66 patients were treated with PRP after hip arthroscopy under ultrasound guidance (hip arthroscopy+PRP group). The two groups were compared before, at 12 months after surgery and at the last follow-up regarding the following items: Visual Analogue Scale (VAS), Modified Harris Hip Score, International Hip Outcome Tool-12 (iHOT-12), and Hip Outcome Score Activities of Daily Living Scale (HOS-ADL). The incidence rate of complications after surgery was compared between the two groups.Results:A total of 108 patients were followed up for 24-36 months [(28.5±3.8)months], while 25 patients were lost to follow-up because of withdrawal of consent, wrong telephone number, etc, including 11 patients (16.4%) in the hip arthroscopy group and 14 patients (21.2%) in the hip arthroscopy+PRP group. The values of VAS in the hip arthroscopy group before, at 12 months after surgery and at the last follow-up were 5.00(5.00, 7.00)points, 3.00(2.00, 3.75)points, and 1.00(0.00, 2.00)points, respectively; the values of Modified Harris Hip Score were 49.00(39.00, 57.00)points, 76.00(69.25, 82.00)points, and 86.00(82.00, 88.00)points, respectively; the values of iHOT-12 were 0.45(0.28, 0.58)points, 0.69(0.58, 0.80)points, and 0.81(0.70, 0.92)points, respectively; the values of HOS-ADL were 0.52(0.42, 0.68)points, 0.87(0.75, 0.93)points, and 0.93(0.86, 0.99)points, respectively. The scores of VAS in the hip arthroscopy + PRP group before, at 12 months after surgery and at the last follow-up were 6.00(5.00, 7.00)points, 3.00(2.00, 3.75)points, and 1.00(0.00, 2.00)points, respectively; the values of Modified Harris Hip Score were 46.50(37.00, 56.75)points, 78.00(72.00, 84.00)points, and 84.50(82.00, 88.00)points, respectively; the values of iHOT-12 were 0.42(0.26, 0.51)points, 0.66(0.58, 0.74)points, and 0.81(0.68, 0.88)points, respectively; the values of HOS-ADL were 0.54(0.38, 0.65)points, 0.87(0.72, 0.96)points, and 0.94(0.86, 1.00)points, respectively. In both groups, VAS, Modified Harris Hip Score, iHOT-12, and HOS-ADL were significantly improved at 12 months after surgery and at the last follow-up compared with those before surgery, and were further improved at the last follow-up compared with those at 12 months after surgery (all P<0.01). There were no significant differences in VAS, Modified Harris Hip Score, iHOT-12 and HOS-ADL between the two groups before, at 12 months after surgery and at the last follow-up (all P>0.05). There was no significant difference in the incidence rates of postoperative hip pain and clicking between the two groups (both P>0.05). Conclusion:Hip arthroscopy can considerably improve short-term hip symptoms and function in FAI patients, but the use of PRP treatment after hip arthroscopy cannot further improve its short-term efficacy in FAI patients.
6.Analysis of the efficacy of arthroscopic transverse release of iliotibial band through peritrochanteric space for the treatment of external snapping hip
Yidong WU ; Kangkang YU ; Zhongyao LI ; Lu GAN ; Qi JIA ; Zhongyuan ZHAO ; Yang HE ; Zhikai GUO ; Chunbao LI
Chinese Journal of Orthopaedics 2024;44(1):18-24
Objective:To analyze the clinical efficacy of arthroscopic transverse release of the iliotibial band through peritrochanteric space for the treatment of external snapping hip.Methods:A total of 30 patients (12 males and 18 females) with bilateral external snapping hip underwent arthroscopic transverse release of the iliotibial band through peritrochanteric space in Department of Sports Medicine, Senior Department of Orthopaedics, the Fourth Medical Center, Chinese PLA General Hospital were retrospectively analyzed from May 2021 and June 2022. The average age was 32.5±8.2 years (range, 17-51 years). At the same time, 30 patients who underwent arthroscopic external release of the iliotibial band through the external surface of the iliotibial band (external iliotibial band group) were selected as control group, including 13 males and 17 females, aged 29.5±6.8 years (range, 11-45 years). The visual analogue scale (VAS), modified Harris hip score (mHHS), and gluteal muscle contracture disability scale (GDS) were compared between the two groups at preoperative, 6 months postoperative, and final follow-up.Results:All patients successfully completed the operation and were followed up for 17.5±3.3 months (range, 12-25 months). The VAS scores of the two groups at the last follow-up were lower than those before operation ( P<0.05). The mHHS scores before operation, 6 months after operation and at the last follow-up in the peritrochanteric space group were 76.5 (67.0, 85.5), 98.5 (94.8, 100.0) and 100.0 (97.0, 100.0), respectively, and those in the external iliotibial band group were 80.5 (70.0, 86.0), 100.0 (96.0, 100.0) and 100.0 (99.5, 100.0). The differences in mHHS scores between the two groups were statistically significant for intragroup comparisons ( P<0.05); of these, 6 months postoperatively and at the last follow-up were greater than preoperatively, with statistically significant differences ( P<0.05); the differences at 6 months postoperatively and at the last follow-up were not statistically significant ( P>0.05). There was no significant difference in mHHS scores between groups at different time points ( P>0.05). The GDS before operation, at 6 months after operation and at the last follow-up were 47.0 (35.8, 64.5), 90.0 (81.0, 94.0) and 93.5 (89.8, 98.0) in the peritrochanteric space group, and 51.0 (38.0, 64.5), 50.0 (81.0, 94.0) and 93.5 (89.8, 98.0) in the external iliotibial band group, respectively. The differences in GDS between the two groups were statistically significant for intragroup comparisons ( P< 0.05); of these, 6 months postoperatively and at the last follow-up were greater than preoperatively, with statistically significant differences ( P<0.05); the differences at 6 months postoperatively and at the last follow-up were not statistically significant ( P>0.05). There was no significant difference in GDS between groups at different time points ( P>0.05). Conclusion:Arthroscopic transverse release of the iliotibial band through peritrochanteric space for the treatment of external snapping hip can effectively reduce hip pain and improve hip function, with satisfactory clinical results, and can be used as an alternative treatment to transverse release through the external surface of the iliotibial band.
7.Optimization of the Extraction Technology of Senecio scandens by Orthogonal Design Combined with Informa- tion Entropy Weighting Method
Yan LI ; Yanfan PAN ; Tao ZHANG ; Zhongyao HAN
China Pharmacy 2020;31(12):1470-1474
OBJECTIVE:To optimize the extraction technology of Senecio scandens ,and to provide reference for the further development and utilization of the medicinal material. METHODS :The method of reflux extraction with 80 ℃ water bath was used to extract S. scandens . HPLC method was adopted to determine the contents of chlorogenic acid and hyperoside. The determination was performed on Diamonsil C 18 with mobile phase consisted of 0.2% glacial acetic acid water solution-methanol (82∶18,V/V)at the flow rate of 1.0 mL/min;the column temperature was set at 35 ℃;the detection wavelength was 327 nm,and the sample size was 5 μL. UV-Vis spectrophotometry was used to determine the contents of total flavonoids(by rutin )and total alkaloids (by matrine)and the detection wavelengths were set at 509 nm and 208 nm,respectively. Based on single factor tests ,using ethanol volume fraction (A,%),solvent folds (B,mL/g),extraction time (C,h),extraction times (D,times)as factors ,using the contents of chlorogenic acid ,hyperoside,total flavonoids and total alkaloids as indexes ,and t he weight coefficients of above indicator components were calculated based on information entropy weighting method so as to calculate their comprehensive scores , then L 9(34)orthogonal design was used to further optimize the extraction technology. The validation tests were also performed. RESULTS:The optimal extraction technology of S. scandens included that 8-fold 50% ethanol,extracting for 3 times,lasting for 1.5 h each time. Results of 3 times of validation tests showed that RSDs of the contents of chlorogenic acid ,hyperoside,total flavonoids and total alkaloids were all lower than 1.5%(n=3). CONCLUSIONS :The optimized technology is reproducible , stable and feasible ,and can be used for the extraction of S. scandens .
8.Analysis of Ongoing Change Characteristics of the Contents of Syringin and Total Flavonoids in Different Medicinal Parts of Toricellia angulata from Guizhou
Zhongyao HAN ; Wei SONG ; Yan LI ; Tao ZHANG ; Wenshuang TANG ; Fujun ZHOU
China Pharmacy 2020;31(17):2124-2128
OBJECTIVE:To study ongoing change characteristics of the contents of syringin and total flavonoids in different medicinal parts (root bark ,tree bark ,leaf)of Toricellia angulata from Guizhou ,and to provide reference for the development and application of T. angulata . METHODS :The root bark ,tree bark and leaf parts of T. angulata during different harvesting periods (Jan.-Dec.) were taken as the research samples. The content of syringin was determined by HPLC. The determination was performed on Agela Promosil C 18 column with mobile phase consisted of 0.5% phosphoric acid solution-acetonitrile (gradient elution)at the flow rate of 1.0 mL/min. The detection wavelength was set as 210 nm,and column temperature was 35 ℃. The sample size was 5 μL. The content of total flavonoids was determined by UV-visible spectrophotometry under detection wavelength of 510 nm. RESULTS :The linear range of syringin and total flavonoids were 0.095 9-1.150 8 mg/mL(r=0.999 6)and 0.072 2- 1.083 0 mg/mL(r=0.999 9),respectively. RSDs of precision ,stability and repeatability tests were all less than 3%(n=6). The average recoveries were 101.74%(RSD=2.36% ,n=6)and 99.63%(RSD=2.19% ,n=6),respectively. During different harvesting periods ,the contents of syringin in root bark ,tree bark ,leaf of T. angulata collected on Aug. ,May and Sept. were the highest,and the contents of total flavonoids in samples collected on Feb. ,Dec. and Sept. were the highest. The contents of syringin in different medicinal parts of T. angulata were in descending order as follows as tree bark >root bark >leaf,and the content of syringin was commonly relatively high in tree bark part ;the content of total flavonoids in different medicinal parts of T. angulata were in descending order as follows as root bark >tree bark >leaf,and the contents of total flavonoids in three medicinal parts was generally low. The content of total flavonoids in root bark was the highest in Feb. of that year ,and the content of syringin in root bark at same month was second only to Aug. of that year ;the content of syringin in tree bark was the highest in May ,and the content of total flavonoids in tree bark at same month was second only to Oct. and Dec. of that year ;the contents of total flavonoids and syringin in leaf were the highest in Sept. of that year. CONCLUSIONS :It is suggested that Feb. is the best time for harvesting root bark ,May for tree bark and Sept. for leaf of T. angulata .
9.Predictive value of preoperative alkaline phosphatase to prealbumin ratio in prognosis and postoperative complications in patients with hepatocellular carcinoma after radical tumor resection
Shengdeng CHEN ; Zhiqiang MOU ; Zhongyao CHEN ; Jian WEN ; Qiu LI
Journal of Clinical Hepatology 2023;39(1):118-127
Objective To explore the predictive value of preoperative alkaline phosphatase to prealbumin ratio (APR) in prognosis and postoperative complications for patients with hepatocellular carcinoma (HCC) after radical tumor resection. Methods A total of 217 HCC patients who underwent radical tumor resection in the Department of Hepatobiliary Surgery of the Affiliated Hospital of Southwest Medical University from January 2013 to August 2021 were retrospectively recruited and their clinical data were statistically analyzed. The X-tile software was used to obtain the optimal cutoff value of APR. The χ 2 test was conducted to analyze association between preoperative APR and other clinicopathological characteristics. The Kaplan-Meier curve was plotted and the Log-rank test was performed to analyze survival of patients. The univariate and multivariate Cox proportional hazards regression models were used to analysis factors affecting the prognosis of HCC patients. The univariate analysis and multivariate Logistic regression were used to identify factors related with postoperative complications. The receiver operating characteristic (ROC) curve was used to determine the predicting value of APR. Results The optimal cutoff value for APR ratio was 0.5 and these 217 patients were divided into the low- and high APR groups (111 vs 106 cases) accordingly. Compared with the low-APR group, the proportion of patients with ALT (> 50 U/L), Alb (< 40 g/L), the CNLC of the III stage, open surgery, liver cirrhosis, multiple tumor lesions, postoperative complication, and major complication were significantly increased in the high-APR patients (all P < 0.05). Moreover, the 1-, 3-, and 5-year OS were 86.0%, 74.9%, and 71.3%, respectively in the low-APR patients, while the numbers were 79.2%, 57.5%, and 47.0%, respectively, in the high-APR patients, indicating that patients in high-APR group had significantly worse OS ( P =0.002). AFP ( HR =1.774, 95% CI : 1.107-2.843, P =0.017), CNLC stage ( HR =2.708, 95% CI : 1.514-4.844, P =0.001), tumor size ( HR =1.696, 95% CI : 1.060-2.714, P =0.028), and APR ( HR =2.022, 95% CI : 1.244-3.285, P =0.004) were all independent risk predictors for OS. The 1-, 3-, and 5-year RFS were 82.3%, 69.4%, and 61.3%, respectively, in the low-APR patients, whereas the numbers were 76.2%, 54.4%, and 44.2%, respectively in the high-APR patients, suggesting that high-APR patients had significantly worse recurrence-free survival ( P =0.016). The CNLC stage ( HR =2.509, 95% CI : 1.423-4.422, P =0.001), tumor size ( HR =1.725, 95% CI : 1.119-2.660, P =0.014), and APR ( HR =1.619: 95% CI : 1.037-2.527, P =0.034) were all independent FRS predictors. Hypertension ( OR =3.09, 95% CI : 1.385-6.893, P =0.006), open surgery ( OR =4.198, 95% CI : 1.779-9.907, P =0.001), liver cirrhosis ( OR =2.376, 95% CI : 1.194-4.729, P =0.014), and APR ( OR =2.151, 95% CI : 1.160-3.986, P =0.015) were all independent risk predictors for the postoperative major complications. The AUC for APR, ALP, a nd PA in predicting the major complications was 0.625 (95% CI : 0.547-0.702), 0.613 (95% CI : 0.534-0.693), and 0.554 (0.474-0.634). Conclusion Preoperative APR could be used to predict prognosis and postoperative major complications of HCC patients after radical tumor resection.
10.Advance of diagnosis and medical treatment for neonatal necrotizing enterocolitis
Luquan LI ; Lu GUO ; Zhongyao ZHANG ; Xiaoyu HU ; Keran LING
Chinese Journal of Applied Clinical Pediatrics 2020;35(11):810-814
Necrotizing enterocolitis (NEC) is one of the acquired intestinal inflammatory disease in neonate, with a high mortality rate.The morbidity and mortality rate of NEC remain at high level although recognition for NEC has been improved in last decades.Understanding of this disease has been furthered in latest years.The research advance of NEC is reviewed in present which is focus on the diagnosis and medical treatment to provide new evidence for further study.