1.Rapid detection of Pseudomonas aeruginosaby loop-mediated isothermal amplification
Kefeng CAO ; Jilu SHEN ; Xuelong WANG
Acta Universitatis Medicinalis Anhui 2017;52(3):450-453
A rapid and sensitive method of loop-mediated isothermal amplification(LAMP) was established to detect Pseudomonas aeruginosa(P.aeruginosa).Three pairs of LAMP primers(inner,outer and ring primers) were designed according to the gbca gene of P.aeruginosa.Since adding hydroxy naphthol blue(HNB) to the reaction system, a positive reaction was indicated by a colorchange before and after the reaction,and was verified by agarose gellectrophoresis.Both LAMP and PCR were applied to detect clinical specimens, the sensitivity and specificity of the detection method were evaluated,and were compared with those of conventional PCR.A LAMP method for detecting P.aeruginosa was successfully established.The LAMP method showed specificity for P.aeruginosa without other bacteria amplification.The established LAMP method in this study enables rapid,sensitive and specific detection of P.aeruginosa,and can be applied for grass roots and small scale laboratories as well as field surveillance.
2.Research on mechanism of effect of total flavonoidsextracted from Polygonum perfoliatum L.onanti-hepatic fibrosis in rats
Houkang CAO ; Ya GAO ; Simao HUANG ; Kefeng ZHANG
Chinese Pharmacological Bulletin 2017;33(9):1303-1308
Aim Based on the effect of total flavonoids extracted from Polygonum perfoliatum L.(TFP) against dimethylnitrosamine(DMN)-induced hepatic fibrosis(HF), to investigate the anti-fibrotic mechanism of TFP.Methods Ninety SD rats were divided into normal group, model group, colchicines(0.1 mg·kg-1) group, and TFP(200, 100, 50 mg·kg-1) group.Except the rats of normal group, other rats were injected intraperitoneally with volume fraction 0.5% DMN solution(2 mL·kg-1) for eight weeks, once every two days.From the first day of modeling, each administration group was given the corresponding dose of drugs to intervene, and the normal group and model group were given an equal volume of solvent, once a day.At the end of the eighth week, the blood and liver tissues were collected.Liver tissue was taken at a fixed position, and the degree of liver tissue was observed by HE staining.The contents of serum ALT, AST, SOD and MDA were measured using colorimetric method;the levels of serum HA, LN, PCⅢand Ⅳ-C were detected using enzyme-linked immunosorbent assay(ELISA);the levels of TNF-α, IL-1β and IL-6 in liver tissues were detected by ELISA;the expression of α-SMA, TGF-β1, p-JAK2 and p-STAT3 were detected by Western blot.Results Compared with the model group, TFP(200, 100, 50 mg·kg-1) could improve the liver tissue lesions, reduce the expression of ALT, AST, HA, LN, PCⅢ, IV-C and MDA, increase SOD activity, reduce the levels of TNF-α, IL-1β and IL-6, and inhibit the expression of α-SMA, TGF-β1, JAK2, STAT3, p-JAK2 and p-STAT3.Conclusion TFP could inhibit DMN-induced HF of rats, which may be involved with antioxidant and inhibiting expression of TGF-β1, JAK2/STAT3 signaling pathway and inflammatory response.
3.Effects of DCP on alcoholic fatty liver disease in rats via anti-inflammation and antioxidation
Ya GAO ; Kelan ZHANG ; Mingli ZHONG ; Houkang CAO ; Simao HUANG ; Kefeng ZHANG
Chinese Pharmacological Bulletin 2016;32(12):1665-1669
Aim To study the effects of Dicliptera chinensis polysaccharide(DCP)on alcoholic fatty liver disease(AFLD)in rats based on anti-inflammation and antioxidation.Methods 60 rats were randomly divid-ed into six groups:control group,model group,silybin group and DCP of high,medium and low dose groups. The control group was fed with normal diet, other groups were fed with high sugar and high fat diet,and given 5% alcohol 5 mL·kg-1 by gavage.The alcohol consistency increased 5%every week until AFLD mod-els in rats were made after 7 weeks.Except control group,other groups were fed with high sugar and high fat diet,and given 35% alcohol 5 mL · kg-1 and DCP.All rats were killed after five weeks,and blood and liver tissues were collected.The activity of alanine aminotransaminase (ALT),aspartate aminotransferase (AST ), alkaline phosphatase (AKP ), triglyceride (TG),total cholesterol (TC ),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cho-lesterol(HDL-C)in serum were detected by using bio-chemical method. The contents of malondialdehyde (MDA),superoxide dismutase (SOD),reduced gluta-thione(GSH)in liver tissues were detected.The con-tents of tumor necrosis factor-α(TNF-α),interleukin-6 (IL-6 )and transforming growth factor-β1 (TGF-β1 ) were determined by enzyme-linked immunosorbent as-say(ELISA)in liver tissues.The liver tissues were ob-tained and histologic analysis was done through HE. Results DCP reduced the activity or content of ALT, AST,AKP,TG,TC,LDL-C,HDL-C,TNF-α,IL-6, TGF-β1 in serum and liver tissues of rats(P<0.05 ), and increased the activity or content of HDL-C,SOD and GSH (P<0.05 ).DCP could remarkably inhibit the NF-κB expression in liver tissues(P<0.01 ).The pathological examination indicated that DCP could ob-viously alleviate the inflammation and fat denaturation of the liver cells.Conclusion DCP can inhibit the de-velopment of AFLD.The mechanism may be related to antioxidation,free radical scavenging, inhibition of lipidperoxidation,anti-inflammation,and inhibition of the TGF-β1 and NF-κB expression.
4.Protective Effects and the Mechanism Study of Water Extract of Rabdosia serra on Hepatic Fibrosis Induced by Carbon Tetrachloride in Rats
Qiongmei XU ; Yuelong LI ; Houkang CAO ; Chun CHEN ; Gang WANG ; Yizhun ZHU ; Kefeng ZHANG
China Pharmacy 2018;29(20):2791-2796
OBJECTIVE:To study the protective effects and the mechanism of Rabdosia serra water extract(RWE)on hepatic fibrosis(HF)induced by carbon tetrachloride(CCl4)in rats. METHODS:Sixty male SD rats were randomly divided into normal group,model group,colchicine group(0.12 mg/kg),and RWE low-dose,medium-dose and high-dose groups(4,8,16 g/kg,by crude drug),with 10 rats in each group. Except for intraperitoneal injection of olive oil for normal group,other groups were given 40% CCl4olive oil solution intraperitoneally to induce HF model. Since the first day of modeling,each treatment group was given relevant medicine (10 mL/kg) intragastrically, while normal group and model group were given constant volume of water intragastrically,once a day,for consecutive 6 weeks. After medication,biochemical process or ELISA were used to determine the contents of ALT,AST,HA,LN,PCⅢ and Ⅳ-C in serum,the activities or contents of SOD,GSH-Px,MDA,TNF-α,IL-6 and IL-1β in liver tissue. Pathological changes of liver tissue in rats were observed by HE staining. The expression of α-SMA and TGF-β1 in liver tissue were detected by Western blot. RESULTS:Compared with normal group,the contents of ALT,AST,LN,HA,PCⅢ and Ⅳ-C in serum,the contents of MDA,TNF-α,IL-6 and IL-1β in liver tissue were all increased significantly in model group (P<0.01);the activities of SOD and GSH-Px in liver tissue were decreased significantly(P<0.01). Liver fibrosis was obvious, and the relative expression of α-SMA and TGF-β 1were increased significantly (P<0.01). Compared with model group,the contents of ALT,AST,HA,LN,PCⅢ and Ⅳ-C in serum as well as the contents of MDA,TNF-α and IL-6 in liver tissue in colchicines group and RWE groups,the contents of IL-1 β in liver tissue of rats in colchicines group,RWE medium-dose and high-dose groups were all decreased significantly (P<0.05 or P<0.01). The activities of SOD and GSH-Px in liver tissue of rats were increased significantly in colchicines group and RWE groups(P<0.05 or P<0.01). The fibrosis degree of liver tissue was significantly reduced, while the relative expression of α-SMA and TGF-β 1decreased significantly (P<0.01). CONCLUSIONS:RWE can protect CCl4-induced HF model rats,the mechanism of which may be associated with regulating lipid metabolism,relieving liver lipid peroxidation injury and anti-oxidative stress response,inhibiting the release of inflammatory factors and the expression of TGF-β1.
5.The impact of apoB/apoA1 on lupus nephritis and renal insufficiency and its diagnostic value
Wei ZHAO ; Bo HUANG ; Kefeng CAO
Journal of Chinese Physician 2024;26(2):223-227
Objective:To analyze the impact and diagnostic value of apoB/apoA1 on lupus nephritis (LN) and its renal dysfunction.Methods:A total of 134 patients diagnosed with systemic lupus erythematosus (SLE) at the Taihe Hospital of Anhui University of Traditional Chinese Medicine from July 2019 to January 2023 were selected and divided into LN group ( n=82) and simple SLE group ( n=52). According to the glomerular filtration rate (e-GFR), LN was divided into a group with normal renal function ( n=42) and a group with renal insufficiency ( n=40). We compared the differences in clinical data between different groups and analyzed the diagnostic value of apoB, apoA1, and apoB/apoA1 for LN and its renal insufficiency. Results:The results of binary logistic regression analysis showed that apoB/apoA1, SLE disease activity index (SLEDAI), anti double stranded DNA (dsDNA), complement 3 (C3), and albumin (ALB) were independent influencing factors for LN ( OR=4.033, 1.179, 3.148, 0.374, 0.879, all P<0.05). The area under the curve (AUC) of apoB, apoA1, and apoB/apoA1 for diagnosing LN were 0.623, 0.662, and 0.742, respectively. The diagnostic efficacy of apoB/apoA1 was higher than that of apoB and apoA1, and the differences are statistically significant (all P<0.05). Regardless of whether confounding factors were adjusted or not, apoB/apoA1 were all risk factors for LN with e-GFR>60 ml/(min·1.73 m 2), e-GFR 30-60 ml/(min·1.73 m 2), and e-GFR<30 ml/(min·1.73 m 2) (all P<0.05). ApoB/apoA1, anti dsDNA, C3, and ALB were all independent influencing factors for renal insufficiency LN ( OR=3.778, 2.669, 0.415, 0.884, all P<0.05). The AUC for diagnosing renal insufficiency LN in apoB, apoA1, and apoB/apoA1 were 0.623, 0.640, and 0.730, respectively. The diagnostic efficacy of apoB/apoA1 was higher than that of apoB and apoA1, and the differences were statistically significant (all P<0.05). Conclusions:ApoB/apoA1 is an independent influencing factor for the occurrence of LN and renal insufficiency LN, and has good diagnostic value for LN and renal insufficiency LN.
6.Non-alcoholic fatty liver disease and liver transplantation
Pengcheng ZHANG ; Kefeng DOU ; Zhaoxu YANG ; Ruohan ZHANG ; Hongtao ZHANG ; Wei PENG ; Yanbing CAO ; Weimin LI
Chinese Journal of Hepatobiliary Surgery 2020;26(2):155-157
Non-alcoholic fatty liver disease (NAFLD) is characterized by increased fat depositions in the liver while the patients do not have drinking history.NAFLD has a prevalence of 10% ~40% in global,25% ~26% in Western populations.From 2004 to 2013,the numbers of new patients on the waitlist who had NASH increased by 170% in America.The prevalence of NAFLD in China is 20%.With the decrease of HBV and HCV and the increase of diabetes mellitus type 2 and obesity,NAFLD will become the most common chronic liver disease in China over the next 20 years.NAFLD related end-stage liver disease will become the most common indication of liver transplantation.In this paper,the epidemiological features,pathogenesis,indication and prognosis of liver transplantation are reviewed.
7.Influencing of high sodium donor liver transplantation from the death of a citizen′s organ donation on the prognosis of recipients
Yang CAO ; Hongtao ZHANG ; Wei PENG ; Qingqiang WANG ; Hengchao YU ; Kaishan TAO ; Kefeng DOU ; Zhaoxu YANG
Chinese Journal of Digestive Surgery 2023;22(2):251-259
Objective:To investigate the influencing of high sodium donor liver transplan-tation from the death of a citizen′s organ donation (DCD) on the prognosis of recipients.Methods:The retrospective cohort study was constructed. The clinicopathological data of 125 pairs of donors and recipients who underwent DCD liver transplantation in Xijing Hospital of Air Force Military Medical University from January 2015 to June 2021 were collected. Of the 125 donors, there were 93 males and 32 females. Of the 125 recipients, there were 92 males and 33 females, aged 48(41,55)years. According to the last time of serum sodium level of donor liver in the 125 recipients, 9 donor livers with serum sodium level ≥170 mmol/L were divided into group 1 (extremely high sodium), 33 donor livers with serum sodium level ≥150 mmol/L and <170 mmol/L were divided into group 2 (moderate high sodium), and 83 donor livers with serum sodium level <150 mmol/L were divided into group 3 (normal sodium), respectively. Observation indicators: (1) postoperative recover situations; (2) follow-up and survival analysis. Measurement data with normal distribution were represented as Mean± SD. Repeated measures were analyzed by repeated measures ANOVA. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was analyzed using the Kruskal-Wallis test. Count data were described as absolute numbers, and Pearson chi-square test or Fisher exact probability were used for data test. The Kaplan-Meier method was used to draw survival curves and Log-rank test was used for survival analysis. Results:(1) Postoperative recover situations. The changes of alanine transaminase (AlT), aspartate aminotransferases (AST), total bilirubin (TBil), alkaline phosphatase (ALP), prothrombin time (PT), international normalized ratio (INR), albumin (Alb) and creatinine (Cr) from the first day to the 14th day after operation were (736±972)IU/L to (75±46)IU/L, (1 290±1 651)IU/L to (38±20)IU/L, (102±98)μmol/L to (33±11)μmol/L, (66±34)IU/L to (104±54)IU/L, (19.9±3.3)seconds to (11.3±1.0)seconds, 1.76±0.31 to 1.00±0.08, (34±5)g/L to (38±3)g/L and (91±41)μmol/L to (76±19)μmol/L, respectively, in the recipients of group 1. The above indicators were (505±377)IU/L to (48±46)IU/L, (855±727)IU/L to (24±17)IU/L, (64±42)μmol/L to (32±22)μmol/L, (68±51)IU/L to (91±46)IU/L, (16.8±3.5)seconds to (11.9±1.2)seconds, 1.47±0.30 to 1.04±0.09, (33±4 g/L) to (40±5)g/L and (106±32)μmol/L to (97±27)μmol/L in the recipients of group 2 and (637±525)IU/L to (65±60)IU/L, (929±1 193)IU/L to (33±27)IU/L, (66±48)μmol/L to (33±36)μmol/L, (64±28)IU/L to (125±64)IU/L, (17.2±4.7)seconds to (13.3±12.8)seconds, 1.51±0.42 to 1.05±0.13, (35±6)g/L to (39±4)g/L, (105±44)μmol/L to (94±40)μmol/L in the recipients of groups. Results of overall effect showed there were significant differ-ences in the change trend of TBil (time effect, inter-group effect, interaction effect) in recipients among the three groups after liver transplantation ( Fgroup=5.42, Ftime=22.78, Finteraction=3.85, P<0.05). There were significant differences in the time effect of ALT, AST, ALP, PT, INR, Alb, Cr in recipients among the three groups after liver transplantation ( Ftime=50.17, 36.24, 19.24, 10.55, 59.61, 41.94, 10.82, P<0.05). (2) Follow-up and survival analysis. All recipients were followed up. Cases with early postoperative liver dysfunction, cases with donor liver failure 1 year after operation, cases with biliary complica-tions 1 year after operation, cases with vascular complications 1 year after operation, cases with rejection 1 year after operation were 2, 1, 0, 0, 0 in the recipients of group 1. The above indicators were 2, 1, 3, 0, 1 in the recipients of group 2 and 10, 8,20, 1, 6 in the recipients of group 3. There was no significant difference in the above indicators among the three groups ( χ2=1.58, 0.60, 5.19, 1.62, 0.97, P>0.05). The 1-year and 3-year cumulative survival rates of the donor liver were 100.00% and 100.00% in the recipients of group 1 after liver transplantation. The above indicators were 94.74% and 77.16% in the recipients of group 2 and 91.57% and 89.30% in the recipients of group 3. There was no significant difference in the cumulative survival rate of donor liver among the three groups ( χ2=2.69, P>0.05). The 1-year and 3-year cumulative survival rates were 100.00% and 100.00% in the recipients of group 1 after liver transplantation. The above indicators were 93.74% and 77.16% in the recipients of group 2 and 89.40% and 86.00% in the recipients of group 3. There was no significant difference in the cumulative survival rate among the three groups ( χ2=1.94, P>0.05). Conclusion:Donor livers with high serum sodium level can be used in the DCD liver transplantation.
8.Prediction Analysis on the Potential Suitable Areas of Zanthoxylum nitidum(Roxb.)DC.in China under Climate Change
Zhiyuan ZHANG ; Mingli ZHONG ; Bo LI ; Kefeng ZHANG ; Houkang CAO
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(6):7-13
Objective To predict and analyze the potential suitable areas of the Guangxi characteristic medicinal plant Zanthoxylum nitidum(Roxb.)DC.in China under climate change;To provide reference for ecological conservation and artificial cultivation of Zanthoxylum nitidum(Roxb.)DC.Methods Based on the 49 pieces of geographical distribution information of Zanthoxylum nitidum(Roxb.)DC.in China,19 environmental factors,and the maximum entropy(MaxEnt)model,a species distribution model was constructed to predict and analyze the potential growth areas of Zanthoxylum nitidum(Roxb.)DC.under current and future climate change.Results Under the current climate change,the potential suitable areas of Zanthoxylum nitidum(Roxb.)DC.in China mainly depended on 4 environmental factors:precipitation in the coldest season(Bio19),precipitation in the warmest season(Bio18),precipitation in the driest season(Bio17),and average annual temperature(Bio1).Under the future climate change,the center of mass of the high suitable areas of Zanthoxylum nitidum(Roxb.)DC.will mainly migrate to the northeast and northwest,and the migration distance will range from 13.63 km to 153.95 km.Compared with the present,the stable and unchanged high suitable areas of Zanthoxylum nitidum(Roxb.)DC.under future climate change will be reduced to about 21.68%,and mainly distributed in southwest Guangxi,and these stable and unchanged areas will become the first choice for semi-artificial planting of Zanthoxylum nitidum(Roxb.)DC.Conclusion The results are helpful to provide scientific basis for formulating reasonable ecological conservation and artificial cultivation strategies to reduce the impact of climate change on the population spread of Zanthoxylum nitidum(Roxb.)DC.