1.Analysis of influence of overweight on appendicitis and laparoscopic appendectomy
Enxu BI ; Quanmiao YU ; Tao XIA ; Ruipeng ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2017;24(8):1235-1238
Objective To analyze the effect of overweight on appendicitis and laparoscopic surgery.Methods Clinical data of 200 patients with laparoscopic appendectomy were analyzed retrospectively.Overweight and non-overweight patients were compared.The risk factors of appendicitis were analyzed.Results The operation time [(56.19 ± 30.42) min vs.(48.40 ± 21.03) min] and length of stay [(5.35 ± 2.04) d vs.(4.52 ± 1.49) d] between the overweight and non-overweight groups had statistically significant differences (t =2.040,P =0.043,t =3.219,P =0.002).Overweight was correlated with appendix inflammation (OR =0.149,P =0.000).Conclusion Over weight patients are prone to develope into complicated appendicitis.It should take surgeon's attention for more difficult operation and more complication.
2.Impact of ischemic preconditioning on dynamics of homing of endothelial progenitor cells after renal ischemia reperfusion injury
Hao LIU ; Ran WU ; Ruipeng JIA ; Bing ZHONG ; Peng YU ; Yan ZHAO
Chinese Journal of Nephrology 2013;(3):199-203
Objectives To investigate the impact of ischemic preconditioning (IPC) on dynamics of homing of endothelial progenitor cells (EPCs) after renal ischemia reperfusion injury (IR).Methods Sixty male Sprague-Dawley rats were randomly divided into three groups after right-side kidney nephrectomy:for sham-operated rats,lumbotomy without vascular clamping was performed; IR rats were clamped renal blood vessels for 40 minutes while IPC rats were pre-treated with 15 min ischemia and 10 min reperfusion.At 3,12,24 h,and 3 days after reperfusion,the pool of circulating,kidneys,lungs and spleens were harvested.The extent of renal injury was assessed by biochemical and histological examination.The dynamics of homing of EPCs was observed by flow cytometry.Results The rats in IPC group exhibited significant improvements in renal function and morphology.Compared with IR group and sham group,the number of EPCs in blood was increased in the IPC group at 12 h and 24 h after reperfusion (P < 0.05).The number of EPCs in kidney was increased at all times pointin the IPC group and IR group as compared to the sham group (P < 0.05.In addition,EPCs number was increased in IPC group compared with the IR group at 12 h and 24 h [(11.36±0.66)% vs (6.37±0.69)%,(6.31±0.70)% vs (4.40±0.60)%,all P< 0.05].Compared with IR group and sham group,the number of EPCs in the lung was increased in the IPC groups at 12 h after reperfusion [(2.95±0.66)% vs (1.78±0.59)%,(1.66±0.61)%,all P < 0.05].The number of EPCs in spleen was increased in the IPC group at 72 h as compared with the IR group and sham group [(0.55±0.06)% vs (0.34±0.07)%,(0.31±0.06)%,all P < 0.05].Conclusions Endogenous EPCs may home to injured kidney after IPC.EPCs can also gather in the lungs and spleen.
3.Study on gene expression level of chondrocytes and cartilage in adult rabbits
Wangping DUAN ; Yongzhuang HAO ; Wenjie SONG ; Ruipeng ZHAO ; Xiaochun REN ; Yu ZHAO ; Qi LI ; Zhenwei SUN ; Pengcui LI ; Xiaochun WEI
Chinese Journal of Rheumatology 2023;27(7):459-463
Objective:To investigate the differences in gene expression levels in knee chondrocytes and chondrons in vitro.Methods:The chondrocytes and chondrons were isolated from full thickness of the 8-months ( n=5) rabbit knees cartilage. Chondrons from right knee were enzymatically isolated using 0.3% dispase and 0.2% collagenase-2 with shaking for 3 hours. Chondrocytes were isolated by 0.4% Pronase and 0.025% collagenase-2 from left knee. The mRNA levels in chondrocytes and chondrons were analyzed by quantitative real-time PCR, including matrix proteins [aggrecan(Agg), collagen(Col-2), Col-6A6, Col-10, Col-11], MMPs and inhibitors (MMP-1, MMP-3, MMP-9, MMP-13, TIMP-1, TIMP-2, TIMP-3), cytoskeletal proteins (Sox-9, vinculin, tubulin, actin), cytokines (IL-β, TNF-α). Analysis was performed using SPSS 16.0 statistical software, and the two-group comparisons were considered as significant by t-test at P<0.05. Results:Compared to the chondrocytes, the Agg [(5.78±0.90) vs (1.89±0.27), t=9.26, P<0.001], Col-2 [(6.29±0.76) vs (3.06±0.60), t=7.46, P<0.001], Col-6A6 [(0.89±0.18) vs (0.22±0.06), t=7.90, P<0.001], Col-10 [(3.83±0.76) vs (1.00±0.26), t=7.88, P<0.001] and TIMP-1 [(1.98±0.85) vs (1.03±0.34), t=2.32, P=0.049], TIMP-2[(3.46±1.50) vs (1.52±1.06), t=2.36, P=0.046], TIMP-3 [(3.96±0.50) vs (1.36±0.18), t=10.94, P<0.001], Sox-9 [(7.09±2.93) vs (3.24±0.77), t=2.84, P=0.022], vinculin [(3.42±1.69) vs (1.46±0.68), t=2.41, P=0.043], tubulin[(9.34±0.71) vs (2.35±0.80), t=14.61, P<0.001] showed higher expression in the chondrons. Compared to the chondrocytes, the MMP-1 [(1.02±0.30) vs (2.67±0.45), t=6.91, P<0.001], MMP-3 [(1.21±0.32) vs (2.52±0.79), t=3.44, P=0.009], MMP-13 [(1.23±0.34) vs (3.42±0.86), t=5.30, P=0.007], IL-1β [(1.02±0.14) vs (2.70±0.49), t=7.37, P<0.001], TNF-α [(0.99±0.08) vs (3.15±0.54), t=8.85, P<0.001] showed lower expression in the chondrons. There were no difference between chondrons and chondrocytes for Col-11, MMP-9, actin ( P>0.05). Conclusion:The gene expression of extracellular matrix components are higher and the gene expression levels of inflammatory factors and MMPs are decreased in chondrons compared with the chondrocytes, suggesting the chondrons have more multiplication potential as seeding cells for tissue-engineered cartilage.
4.Establishment of Specific Chromatogram and TLC Identification for Qingxin Lianziyin
Wenya GAO ; Xiujing MA ; Chang GAO ; Haiyu ZHAO ; Yanyan ZHOU ; Hongjie WANG ; Ruipeng YU ; Yipeng ZHAO ; Cuie YAN ; Lifang GAO ; Nan SI ; Baolin BIAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(11):34-42
ObjectiveTo establish the specific chromatogram and thin layer chromatography(TLC) of Qingxin Lianziyin(QXLZY) benchmark samples, in order to clarify the key quality attributes and provide a reference for the quality evaluation of QXLZY. MethodHigh performance liquid chromatography(HPLC) specific chromatogram of QXLZY benchmark samples was developed by using a YMC Hydrosphere C18 column(4.6 mm×250 mm, 5 μm) with the mobile phase of acetonitrile(A)-0.2% formic acid aqueous solution(B) for gradient elution(0-10 min, 5%-20%A; 10-20 min, 20%A; 20-25 min, 20%-24%A; 25-40 min, 24%-30%A; 40-55 min, 30%-50%A; 55-65 min, 50%-100%A; 65-75 min, 100%A; 75-75.1 min, 100%-5%A; 75.1-90 min, 5%A), and the detection wavelength was 360 nm. Ultra-high performance liquid chromatography-linear ion trap/orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) with electrospray ionization(ESI) was used to identify the components of QXLZY benchmark samples by accurate relative molecular weight and multilevel MS fragment ion information, the detection conditions were positive and negative ion modes and data dependency scanning mode. TLC identification methods for Ophiopogonis Radix, Lycii Cortex, Nelumbinis Semen, Poria, Astragali Radix and Ginseng Radix et Rhizoma in QXLZY were established. ResultA total of 15 characteristic peaks were identified from Glycyrrhizae Radix et Rhizoma, Plantaginis Semen and Scutellariae Radix, and the relative standard deviations of the retention times of 15 characteristic peaks in 15 batches of QXLZY benchmark samples were≤3% with peak 8(baicalin) as the reference peak. A total of 100 compounds, including flavonoids, organic acids, saponins, amino acids and others, were identified in the benchmark samples by UHPLC-LTQ-Orbitrap MS. The established TLC had good separation and was suitable for the identification of Ophiopogonis Radix, Lycii Cortex, Nelumbinis Semen, Poria, Astragali Radix and Ginseng Radix et Rhizoma in QXLZY. ConclusionThe material basis of QXLZY benchmark samples is basically determined by MS designation and source attribution. The established specific chromatogram and TLC of QXLZY are simple, stable and reproducible, which can provide a reference for the development and quality control of QXLZY.
5.Establishment of Specific Chromatogram and Thin-layer Identification Method for Kaixinsan
Wenya GAO ; Min FENG ; Chang GAO ; Haiyu ZHAO ; Yanyan ZHOU ; Hongjie WANG ; Ruipeng YU ; Yipeng ZHAO ; Lili ZHANG ; Yanqing GAO ; Yanhong XIAO ; Cuina CHENG ; Xiaolu WEI ; Nan SI ; Baolin BIAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(13):37-44
ObjectiveTo establish the specific chromatogram and thin layer chromatography(TLC) identification method of Kaixinsan(KXS) samples, in order to clarify the key quality attributes and provide reference for the quality evaluation of KXS. MethodHigh performance liquid chromatography(HPLC) specific chromatogram of KXS was developed with YMC Hydrosphere C18 column(4.6 mm×250 mm, 5 μm), the mobile phase was acetonitrile(A)-0.2% formic acid aqueous solution(B) for gradient elution(0-15 min, 2%-20%A; 15-25 min, 20%-25%A; 25-30 min, 25%-30%A; 30-45 min, 30%-31%A; 45-50 min, 31%-44%A; 50-65 min, 44%-45%A; 65-73 min, 45%-75%A; 73-95 min, 75%-100%A; 95-105 min, 100%A; 105-105.1 min, 100%-2%A; 105.1-120 min, 2%A), the detection wavelength was 320 nm. Ultra high performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) was used to identify the chemical components of KXS with electrospray ionization(ESI), negative ion mode and scanning range of m/z 50-2 000. TLC identification methods for Poria and Ginseng Radix et Rhizoma in KXS were established. ResultThere were 11 common peaks in the specific chromatogram of KXS, attributed to Polygalae Radix, Poria and Acori Tatarinowii Rhizoma. Taking peak 9(α-asarone) as the reference peak, the relative standard deviations of the retention times of 15 batches of KXS samples were<0.2%. A total of 34 compounds were identified by UHPLC-LTQ-Orbitrap MS, including terpenoids, phenylpropanoids, oligosaccharides and ketones. The established TLC had good separation and was rapid, reliable, simple, feasible, suitable for the identification of Poria and Ginseng Radix et Rhizoma in KXS. ConclusionThe specific chromatogram and TLC of KXS are stable and reproducible. The material basis of KXS is basically clarified by MS, which can provide a reference for the development and quality control of KXS.