1.Exploring Mechanism of Different Processed Products of Baiyaojian and Its Formula in Treating Ulcerative Colitis Based on Serum Metabolomics and Flow Cytometry
Ruisheng WANG ; Yafang HOU ; Wenwen CAO ; Yahong ZHAO ; Zhenling ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):172-181
ObjectiveTo explore the potential mechanism of different processed products of Baiyaojian and its compound Xiangmei pills in rats with ulcerative colitis(UC) by comparing the pharmacodynamic and metabolomic differences. MethodEighty SD rats were acclimatized and kept for 3 d, and randomly divided into 8 groups[blank group, model group, mesalazine group(0.4 g·kg-1), Baiyaojian group(1.89 g·kg-1), stir-fried Baiyaojian group(1.89 g·kg-1), carbonized Baiyaojian group(1.89 g·kg-1), and Xiangmei pills low and high dose groups(1.89, 5.67 g·kg-1)], with 10 rats in each group. Rats in the blank group were administered physiological saline by gavage, and rats in the remaining 7 groups were orally administered 5% dextran sodium sulfate(DSS) solution daily for 8 consecutive days to induce UC model. After successful modeling, each dosing group was given the corresponding dose of drug solution by gavage, and the blank and model groups were given equal amounts of saline by gavage, and the drug was administered continuously for 8 d. Then serum levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, IL-10 and IL-1β were measured by enzyme-linked immunosorbent assay(ELISA), hematoxylin-eosin(HE) staining was used to observe the histopathological changes of colon tissue, the proportion of T helper 17 cells(Th17) and regulatory T cells(Treg) in the peripheral blood of rats in each group was detected by flow cytometry. The endogenous metabolites in serum of rats were detected by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS), and the differential metabolites were characterized by combining principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA), and were analyzed according to the variable importance in the projection(VIP) value>1.0 and P<0.05, and potential metabolic pathways were analyzed according to Human Metabolome Database(HMDB). ResultCompared with the blank group, the colon tissue of the model group was congested and the mucosa was ulcerated, the colon length was significantly reduced(P<0.01) and the quality was significantly increased(P<0.05), the proportion of Th17/Treg in the peripheral blood and the serum levels of TNF-α, IL-6 and IL-1β were significantly increased, while the IL-10 expression wassignificantly reduced(P<0.05, P<0.01). Compared with the model group, the colon tissue of UC rats in each treatment group was improved with scattered ulcers, reduced inflammatory cell infiltration, significantly increased colon length, and significantly decreased mass(P<0.05), the proportion of Th17/Treg in the peripheral blood decreased, the expression of TNF-α,IL-6 and IL-1β was significantly reduced(P<0.05, P<0.01), while the IL-10 expression was significantly increased(P<0.01). The therapeutic effect of different administration groups on UC was in the order of high dose group of Xiangmei pills>low dose group of Xiangmei pills>carbonized Baiyaojian group>stir-fried Baiyaojian group>Baiyaojian group. And a total of 26 differential metabolites were screened in the metabolomics results. Compared with the blank group, 14 differential metabolites were up-regulated and 5 metabolites were down-regulated in the model group, and 14, 9, 14, 12 and 17 metabolites could be recalled in the Baiyaojian group, stir-fried Baiyaojian group, carbonized Baiyaojian group, Xiangmei pills low and high dose groups. The main metabolic pathways involved were citrate cycle pathway, pantothenic acid and coenzyme A biosynthesis pathway, aromatic hydrocarbon receptor(AhR) signaling pathway, glycolysis/gluconeogenesis pathway. ConclusionThe therapeutic effect of Baiyaojian on UC is significantly improved after charcoal stir-frying, and the efficacy is more prominent when combined with Angelicae Dahuricae Radix and Mume Fructus Carbonisata, which can provide a basis for the development of Baiyaojian compound preparations.
2.Comparison of Effect of Ancient and Modern Processing Methods on Chemical Components of Lilii Bulbus Decoction Based on UHPLC-Q-Orbitrap HRMS
Yongqi ZHAO ; Hongwei ZHANG ; Zhenling ZHANG ; Yiming WANG ; Dehua LI ; Mengmei SUN ; Yake ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(7):177-184
ObjectiveTo compare the effects of different processing methods in ancient and modern times on the chemical components of Lilii Bulbus decoction, and to provide experimental support for the origin processing, decoction piece processing and clinical application of this herb. MethodUltra high performance liquid chromatography tandem quadrupole electrostatic field orbitrap high resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) was used for structural identification of the compounds using excimer ions, secondary MS and characteristic fragment ions, and referring to relevant literature and database information. Principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used to screen the main differential components, the differential components were quantitatively studied by high performance liquid chromatography(HPLC), in order to compare the types and contents of chemical components in the decoction of different processing products of Lilii Bulbus. ResultA total of 24 chemical components were identified from the decoction of different processed products of Lilii Bulbus, water extract and scalding liquid of fresh Lilii Bulbus, including 17 phenols, 5 saponins and 2 alkaloids. Compared with the fresh Lilii Bulbus decoction, the contents of regaloside A, p-coumaric acid, colchicine and other components in the decoction of dry Lilii Bulbus processed by scalding method decreased, the content of regaloside C in the decoction of dry Lilii Bulbus processed by steaming method decreased, and the contents of regaloside A and regaloside C in the decoction of fresh Lilii Bulbus processed by water immersion also decreased. Compared with the decoction of dry Lilii Bulbus processed by scalding method, the overall content of components in the fresh Lilii Bulbus decoction and the decoction of fresh Lilii Bulbus processed by water immersion was higher, the contents of components in the decoction of dry Lilii Bulbus processed by steaming method was higher, except for the slightly lower content of regaloside C. ConclusionDifferent processing processes have a certain effect on the types and contents of chemical components in Lilii Bulbus decoction. Scalding process is beneficial to the preservation of Lilii Bulbus, but can cause the loss of effective components. Compared with scalding method, steaming method can prevent browning of Lilii Bulbus and reduce the loss of its active ingredients. The processing method of removing foam after overnight immersion proposed by ZHANG Zhongjing may be more conducive to the treatment of Baihe disease, which can provide reference for the clinical rational application and mechanism research of different processed products of Lilii Bulbus.
3.Optimization of the pressurized processing technology of Strychnos nux-vomica boiled with mung beans based on genetic algorithm
Yiming WANG ; Hongwei ZHANG ; Hongwei ZHANG ; Zhenling ZHANG ; Yongqi ZHAO
China Pharmacy 2023;34(19):2333-2338
OBJECTIVE To optimize the pressurized processing technology of Strychnos nux-vomica boiled with mung beans. METHODS The least squares method was used to establish a one-dimensional model for the effects of four factors, namely, processing time, processing pressure, mung bean dosage and water added, on the contents of strychnine and toxiferine, and the multivariate model hypothesis was proposed by analyzing the function of one-dimensional model. Based on the orthogonal experiment, the genetic algorithm was used to solve the undetermined coefficients in the model. A bi-objective optimization model based on strychnine and toxiferine content was constructed according to the actual conditions, and the optimal technology was obtained by solving the model function and validated. RESULTS The optimal processing technology was boiling S. nux-vomica with mung beans at 2.393 MPa saturated steam pressure for 5.5 h, and then draining; rinsing to remove mung beans, scraping off the bark of S. nux-vomica and cutting into slice of 0.6 mm; using 180 g of mung beans and 15 L of water per 500 g of S. nux- vomica. CONCLUSIONS The optimized pressurized processing technology is stable and feasible, and can provide a reference for the optimization of processing technology of S. nux-vomica boiled with mung beans.
4.Effects of processing and decoction methods on the components of Huatan qushi huoxue decoction
Minghao LIU ; Yasen HUANG ; Zhenling ZHANG ; Lihui ZHANG ; Sutong LIU ; Wenxia ZHAO
China Pharmacy 2022;33(16):1956-1961
OBJECTIVE To establish the fingerprint of Huatan qushi huoxue decoction (HQHD),and to explore the effects of processing and decoction methods on its components. METHODS Using salvianolic acid B as reference ,HPLC fingerprints of 10 batches of single and mixed decoction of crude drugs ,single and mixed decoction of processed products (original formula referred to 8 ingredients were crude drugs ;processed formula referred to processed products of Alisma orientale ,Salvia miltiorrhiza , Curcumae Radix and Bupleuri Radix ,and crude drugs of other ingredients ;single decoction referred to the mixing of each ingredient after being decocted separately ;mixed decoction refers to decocting after mixing all ingredients )were stablished by Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine (2012 edition). The similarity evaluation,common peak identification and attribution ,chemical pattern recognition analysis were also carried out. RESULTS There were 37 common peaks in each fingerprint of 10 batches of single and mixed decoction of crude drugs ,single and mixed decoction of processed ,the similarities with control fingerprint were higher than or close to 0.950. Nine common peaks were identified,i.e. rutin (peak 12),hesperidin(peak 13),salvianolic acid B (peak 16),quercetin(peak 20),silybin(peak 22), luteolin(peak 23),autrantio-obtusin(peak 29),23-acetylalismol C (peak 34),saikosaponin b 2(peak 35);decoction pieces of 8 ingredients all contributed to the fingerprints of HQHD. Principal component analysis (PCA)showed that the 4 kinds of HQHD samples were grouped into one category ,respectively. The clustering result of partial least squares-discriminant analysis was consistent with that of PCA. Corresponding components of peak 1,15,17,18 and 36,salvianolic acid B and luteolin m ay be the differential markers of the quality for mixed decoction samples of crude drugs and processed products ; corresponding components of peak 1,7,17-19,salvianolic acid B and hesperidin may be the differential markers of the quality for single decoction samples of crude drugs and processed products;corresponding components of peak 1,17-19,36, salvianolic acid B and luteolin may be the differential markers of the quality for single decoction and mixed decoction samples of crude drugs ;corresponding components of peak 7,17-19,21, hesperidin,salvianolic acid B ,rutin,luteolin and autrantio-obtusin may be the differential markers of the quality for single decoction and mixed decoction samples of processed products. CONCLUSIONS The established fingerprint of HQHD is stable and reliable. The quality differential components of different decoction samples are luteolin ,hesperidin,etc. The quality differential components of samples processed or not are rutin ,hesperidin,autrantio-obtusin,etc.
5.Effects of miR-369-3p on the proliferation and apoptosis of bladder cancer cells by regulating the expression of VEGFC gene
Zhenling ZHAO ; Huanjun SHAO ; Lina HAO ; Jun LIU
Journal of Chinese Physician 2018;20(1):92-95,99
Objective To observe the effect of miR-369-3p on vascular endothelial growth factor C (VEGFC) gene in bladder cancer cells EJ and 5637 and observe its effect on cell proliferation and apoptosis.Methods Bladder cancer cell lines EJ and 5637 were transfected with miR-NC (control group) or transfected with miR-369-3p (experimental group).The target gene of miR-369-3p was predicted by Targets can target gene prediction software.Fluorescence real-time quantitative polymerase chain reaction (qPCR) was used to detect the changes of VEGFC at mRNA level.The changes of VEGFC,p-Raf-1 and phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) at the protein level were detected by Western blotting.The effect of miR-369-3p on the proliferation of bladder cancer cells was detected by cell counting kit (CCK-8) and clone formation experiment.The effect of miR-369-3p on apoptosis was detected by flow cytometry.Results After transfection with miR-369-3p,the expression of VEGFC at mRNA and protein levels was significantly decreased (P < 0.05),the expression of p-Raf-1 and p-ERK1/2 protein was significantly decreased,the cell proliferation ability was significantly decreased (P < 0.05),the number of clones formed was significantly decreased (P < 0.05) and the apoptosis was significantly increased (P < 0.01).Conclusions miR-369-3p can inhibit the proliferation and promote the apoptosis of bladder cancer cells by interfering with the expression of VEGFC gene,which may provide a new target for the biological therapy of bladder cancer.
6.Application of simethicone before colonoscope examination
Zhenling YANG ; Miao ZHAO ; Qiren HUANG ; Pingguang LEI ; Fen ZHU
China Journal of Endoscopy 2017;23(4):26-29
Objective To explore the application effect of simethicone combined with compound polyethylene glycol electrolyte powder before colonoscope examination. Methods 106 cases underwent colonoscope examination from October 2013 to December 2015 were enrolled in the study. Then all the cases were divided into 2 groups randomly, each with 53 cases. Patients in the control group were treated with simple oral administration of compound polyethylene glycol electrolyte powder, patients in observation group were combined using simethicone and polyethylene glycol electrolyte powder. The preoperative bowel preparation score, lesion detection and the changes of liver and renal function and electrolyte in the two groups were recorded respectively. Results Patients in the observation group were better than the control group in both the bowel preparation score and the detection rate (P < 0.05). There were no obvious adverse reactions in the course of the two groups. Conclusions Simethicone can eliminate intestinal bubbles, and combined use with polyethylene glycol electrolyte powder before colonoscope examination can significantly improve intestinal cleaning effect, improve the colonoscope examination image, enhance the detection rate of lesions.
7.Prenatal diagnosis and pregnancy outcomes in 42 fetuses with pleural effusion
Qinghua WU ; Xiyang MA ; Huirong SHI ; Xiangdong KONG ; Huina LIU ; Zhenling WEI ; Nan BAI ; Junhong ZHAO ; Ruonan ZHU ; Shumin REN ; Ning LIU ; Qiaoling BAI
Chinese Journal of Perinatal Medicine 2017;20(7):521-526
Objective To investigate the value of prenatal diagnosis in identifying the etiology and predicting the prognosis of fetal pleural effusion (FPE).Methods Forty-two cases of FPE were recruited in this study from January 2012 to September 2016.Ultrasound scan and genetic tests were performed on all fetuses.Seven fetuses with severe FPE were given pleurocentesis.Pregnancy outcomes of all the fetuses were followed up.Results FPE was commonly accompanied with other abnormalities,such as ascites,hydrops,hydramnion,hygroma colli,abnormal posturing,joint contractures,arrhythmia and micromandible.Chromosomal abnormality was detected in 11 fetuses (26.2%),of which ten were further confirmed by karyotype analysis,including six with 45,X,three trisomy 21 and one trisomy 18,and one was detected with a 9.83 Mb uniparental disomy (UPD) located at 12q24.21q24.31 by gene chip.One fetus was diagnosed with--SEA/--SEA thalassemia.All of the 12 families decided to terminate the pregnancies after genetic counseling.Among the other 30 fetuses,seven with severe FPE and normal karyotype underwent pleurocentesis.Five of the seven cases were with favorable outcomes,one with progressive hydrops was aborted and one neonate with severe hydrops died after birth.Spontaneous regression of FPE with good outcome was found in two cases.Parents of the other 21 fetuses chose to terminate the pregnancies.Conclusions Prenatal diagnosis is important to identify the etiology and predict the outcome of FPE.Chromosomal abnormality is a relatively common cause of FPE,and 45,X and trisomy 21 are the most common abnormalities.Intrauterine intervention is beneficial for FPE without chromosomal or other definite genetic abnormalities.Genetic test may be of great value for pregnant counseling.
8.MMACHC gene mutation analysis in the prenatal diagnosis of methylmalonic aciduria with homocystinuria
Aojie CAI ; Ya′nan ZONG ; Ning LIU ; Zhenling WEI ; Ying BAI ; Zhenhua ZHAO ; Xiangdong KONG
Chinese Journal of Laboratory Medicine 2016;39(8):613-617
Objective To analyse MMACHC mutations for 45 pedigrees with combined methylmalonic aciduria and homocyctinuria by Sanger sequencing, and to discuss the utility of prenatal genetic diagnosis for these pedigrees.Method Peripheral blood was collected from 45 probands and their parents from 2012-2015 in Genetic Counselling Clinic of the First Affiliated Hospital of Zhengzhou University, and the DNA were extracted from the blood.Then the coding sequence of MMACHC gene was amplified by PCR, and the PCR products were further sequenced to detect mutations for each pedigree.For 12 families, chorionic villus sampling was performed on the pregnant women to make prenatal genetic diagnosis.Result There were 14 distinct mutations detected in the 45 pedigrees, and the most frequent mutations are c.609G>A(W203X),c.658-660delAAG(K220del)and c.80A>G (Q27A).Two of those mutations have not been reported before:one is a splicing site mutation c.81+1G>A;while the other is a missense mutation c.665A>G,p.Y222C.Most mutations were found in exon 4.Among the 12 pedigrees who received prenatal diagnosis, 2 fetuses were normal, 7 fetuses were carriers of heterozygous mutation, and the other 3 fetuses were patients with compound heterozygous mutation or homozygous mutation.The couples whose fetuses were normal or carriers continued the gestation, while the couples whose fetuses were patients decided to terminate the pregnancy.After delivery, the outcome of the fetuses was the same as the prenatal diagnose results.Conclusion Two novel mutations of MMACHC were identified and prenatal genetic diagnosis helps to avoid the delivery of combined methylmalonic aciduria and homocyctinuria patients.
9.Preparation and evaluation of enrofloxacin microspheres and tissue distribution in rats.
Fan YANG ; Jijun KANG ; Fang YANG ; Zhensheng ZHAO ; Tao KONG ; Zhenling ZENG
Journal of Veterinary Science 2015;16(2):157-164
New enrofloxacin microspheres were formulated, and their physical properties, lung-targeting ability, and tissue distribution in rats were examined. The microspheres had a regular and round shape. The mean diameter was 10.06 microm, and the diameter of 89.93% of all microspheres ranged from 7.0 microm to 30.0 microm. Tissue distribution of the microspheres was evaluated along with a conventional enrofloxacin preparation after a single intravenous injection (7.5 mg of enrofloxacin/kg bw). The results showed that the elimination half-life (t(1/2beta)) of enrofloxacin from lung was prolonged from 7.94 h for the conventional enrofloxacin to 13.28 h for the microspheres. Area under the lung concentration versus time curve from 0 h to infinity (AUC(0-infinity)) was increased from 11.66 h.microg/g to 508.00 h.microg/g. The peak concentration (Cmax) in lung was increased from 5.95 microg/g to 93.36 microg/g. Three lung-targeting parameters were further assessed and showed that the microspheres had remarkable lung-targeting capabilities.
Animals
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Anti-Bacterial Agents/*adverse effects
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Drug Delivery Systems/instrumentation/*methods
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Female
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Fluoroquinolones/*adverse effects
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Half-Life
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Humans
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Injections, Intravenous
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Lung/*drug effects
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Male
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*Microspheres
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Rats
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Rats, Sprague-Dawley
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Tissue Distribution
10.Effect of modified closed negative pressure drainage on abnormal healing of abdominal incision
Yanni ZHU ; Lanju DONG ; Dongmei ZHAO ; Yanling BAI ; Zhenling CAI
Modern Clinical Nursing 2014;(6):67-69
Objective To explore the curative effect of modified closed negative pressure drainage on abnormal abdominal incision. Methods Sity-three patients with abnormal healing of abdominal incision in our hospital from January 2012 to June 2013 were selected as the observation group. Another 50 patients from January to December 2011 were assigned to the control group. The former was treated after debridement with human recombinant surface growth factor and modified closed negative pressure drainage,while the latter after debridement with anti-infection and dressings of ethacridine or gentamicinsolution as well as with infrared therapy. The two groups were compared in terms of the frequency,time and cost of changed dressings,and the healing time of the incisions.Results There was insignificant difference in the cost of changed dressings between the two groups(P>0.05). However,the differences were statistically significant in terms of the frequency and time of changed dressings and the healing time of the incisions(P<0.05). The observation group was significantly superior to the control group.Conclusions Modified closed negative pressure drainage is effective in accelerating the healing in abnormal abdominal incisions,reducing the frequency and the time of changed dressings,and easing the economic burdens of the patients,which is suggested to be popularized and applicable in the pimary hospitals.

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