1.Effect of Xuanfei Zhisou Prescription on IL-17 Signaling Pathway in COPD Model Rats
Yanli SHI ; Yunchao WANG ; Zhen YANG ; Xinhua WANG ; Zhiyong JIA
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):28-39
ObjectiveTo investigate the effect of Xuanfei Zhisou prescription on the interleukin-17 (IL-17) signaling pathway in model rats with chronic obstructive pulmonary disease (COPD). MethodA total of 60 Wistar rats were randomly divided into a blank group (10 rats) and a model group (50 rats), and COPD model rats were established by tracheal infusion of lipopolysaccharide combined with passive fumigation. After modeling, the rats were divided into the model group, dexamethasone group, and high, medium, and low-dose Xuanfei Zhisou prescription groups (3.6, 1.8, 0.9 g·kg-1·d-1) according to the random number table. Rats in the blank group and model group were given normal saline of 10 mL·kg-1·d-1 by gavage administration, and the intervention groups of Xuanfei Zhisou prescription were given corresponding drugs. Rats in the dexamethasone group were given dexamethasone of 2.57×10-4 g·kg-1·d-1 for 28 days. The level of pulmonary function indexes in rats was measured by a pulmonary function detector. The contents of interleukin-6 (IL-6), interleukin-8 (IL-8), IL-17, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The positive expressions of IL-17A, IL-17RA, nuclear factor-κB activator 1 (Act1), tumor necrosis factor-associated factor 6 (TRAF6), p-p38 mitogen-activated protein kinase (p-p38 MAPK), nuclear factor-κB p65 (NF-κB p65), and phosphorylation were detected by immunohistochemistry (IHC). The protein expression levels of IL-17A, IL-17RA, Act1, and TRAF6 in the lung tissue were detected by Western blot. The mRNA expressions of IL-17A, IL-17RA, Act1, and TRAF6 in the lung tissue were detected by Real-time polymerase chain reaction (Real-time PCR). ResultCompared with the blank group, the serum contents of IL-6, IL-8, IL-17, IL-1β, and TNF-α in the model group were significantly increased (P<0.05), and the flow rate and volume indexes of pulmonary function in the model group were significantly decreased (P<0.05), while the time indexes and other indexes were significantly increased (P<0.05). The mRNA and protein expression levels of IL-17A, IL-17RA, Act1, and TRAF6 in pulmonary tissue and the positive expressions of downstream NF-κB p65, p-NF-κB p65, and p-p38 MAPK were increased (P<0.05). Compared with the model group, the levels of IL-6, IL-8, IL-17, IL-1β, and TNF-α in the serum of all treatment groups were decreased to varying degrees (P<0.05), and the indexes of pulmonary function were improved to different degrees (P<0.05). The mRNA and protein levels of IL-17A, IL-17RA, Act1, and TRAF6 and the positive expression of downstream NF-κB p65, p-NF-κB p65, and p-p38 MAPK in high and medium-dose Xuanfei Zhisou prescription groups were significantly decreased (P<0.05). ConclusionXuanfei Zhisou prescription can effectively resist inflammation of COPD rats. The mechanism may be related to down-regulating the protein expression of IL-17A, IL-17RA, Act1, and TRAF6, inhibiting downstream NF-κB and p38 MAPK signaling pathways, and reducing the release of IL-6, IL-8, TNF-α, IL-17, and IL-1β, thus reducing the airway inflammation response.
2.Establishment of the Microwave Processing Technology of Yellow Wine-processed Curculigo orchioides and Comparison with Traditional Technology
Yuanyuan LI ; Wei WANG ; Chengguo JU ; Tianzhu JIA
China Pharmacy 2021;32(18):2223-2229
OBJECTIVE:To establish the microwave processing technology of yellow wine-processed Curculigo orchioides , and compare it with traditional technology. METHODS :HPLC method was adopted to determine the contents of curculigoside , orcinol glucoside and orcinol gentiobioside in C. orchioides . Based on the single factor tests ,microwave processing technology was optimized and validated with orthogonal test combined with comprehensive weighted scoring method ,with the amount of yellow wine,microwave power ,wetting time and microwave time as factors ,using the contents of curculigoside ,orcinol glucoside , orcinol gentiobioside and ethanol soluble extract as the indexes. The contents of C. orchioides decoction pieces and processed products were compared. RESULTS :The optimal microwave processing technology included that the amount of yellow wine was 20%(the weight of C. orchioides decoction pieces was 20%),microwave power was 300 W,wetting time was 3 h,microwave time was 2 min. After 3 times of validation tests ,average contents of curculigoside,orcinol glucoside ,orcinol gentiobioside and ethanol soluble extract were 0.095 6%,0.723 9%,0.406 6%,10.115 3%,and RSD were 0.71%,0.54%,0.99%,1.44%(n=3). Average comprehensive score were 99.08(RSD=0.69%,n=3). Except for the content of ethanol soluble extract in traditional wine-processed product ,the contents of curculigoside and orcinol gentiobioside in traditional wine-processed product and microwave processed product as well as the content of ethanol soluble extract in microwave processed product were all significantly higher than C. orchioides decoction pieces ;the contents of curculigoside and orcinol gentiobioside in microwave processed product were both significantly higher than traditional wine-processed product (P<0.05). The contents of orcinol glucoside in 2 processed product were significantly lower than C. orchioides decoction pieces ,while the microwave processed product was significantly higher than traditional wine-processed product (P<0.05). CONCLUSIONS :Optimized microwave processing technology is stable and feasible ,and can be used for the processing of yellow wine-processed C. orchioides .
3.Analysis of cognition, attitude and barriers to community pharmaceutical care among family doctor team members in Shanghai suburb
Libo SHI ; Tianzhu ZHU ; Mei SHEN ; Zhongzhuang WANG
Journal of Pharmaceutical Practice 2021;39(6):577-580
Objective To evaluate the cognition, attitude, and barriers of family doctor team members in chongming district of Shanghai to pharmacists joining the team and providing community pharmaceutical care. To provide the reference resources for the establishment of community pharmaceutical care management mode with appropriate suburban characteristics. Methods In a cross-section study conducted in 2020, an online questionnaire was provided to family doctor teams in 18 townships in Chongming District through group WeChat. Descriptive statistical data were used to analyze the cognition, attitude and barrier of family physician team members to community pharmaceutical care. Results Among the 555 participants in the study, 351 (63.24%) were female, 187 general practitioners (33.69%), 226 nurses (40.72%), and 142 public health physicians (25.59%). There were statistically significant differences in CPC cognition among the three classes of family doctor team members (P<0.05). 126 nurses (51.22%) and 84 public health physicians (68.85%) claimed never heard of CPC. 11.48% public health physicians and 23.58% nurses were familiar with the work content and responsibilities of community clinical pharmacists. General practitioners showed relatively high proportion of 34.76%. 34.22% of general practitioners held a "disagree attitude" against that "community pharmaceutical care can improve the medication efficacy for patients". "Insufficient investment in the health sector" and "insufficient community pharmacists" were the main obstacles to the development of community pharmaceutical care. Conclusion The attitude of family doctors in Chongming area to community pharmaceutical care was conservative. Public healthcare persons and nurses had a low awareness to community pharmaceutical care. The development of community pharmaceutical care was limited by the lack of financial investment and manpower.
4.Effect of pre-implanted anti-adhesion patch in a Miles' operation on the incidence of parastomal hernia in elderly patients
Haisheng LIANG ; Lei ZHANG ; Shuai WANG ; Zengliang LIU ; Tianzhu ZHANG ; Peng LI ; Yue CHANG
Chinese Journal of Geriatrics 2019;38(7):775-778
Objective To investigate the effect of pre-implanted anti-adhesion patch under laparoscopic approach in a Miles' operation on the incidence of parastomal hernia in elderly patients.Methods A total of 68 patients with low rectal cancer admitted into our general surgery department were enrolled in the study from January 2012 to January 2015.All patients received laparoscopic assisted Miles operation.According to whether or not the pre-implanted anti adhesion patch was used during the operation,the 68 cases were divided into pre-implanted patch group(n=38)and the non-implanted patch group(n=30).The operation states,postoperative complications,complications during follow-up and 3-year survival rate were compared between the two groups.Results The operation duration was longer in the pre-implanted patch group than in the non-implanted patch group[(235.7± 33.6)min vs.(94.3±28.9)min,t =5.36,P=0.000].The incidence of complications was lower in the pre implanted patch group than in the non-implanted patch group at 3 years after operation[10.5% (4/38)vs.90.0%(27/30),x2 =42.69,P =0.000].The local recurrence rate,distant metastasis rate and 3-year overall survival rate had no significant difference between the two groups[23.7 % (9/38)vs.23.3% (7/30),31.6% (12/38) vs.33.3% (10/30) and 71.1% (27/38) vs.70.0% (21/30)respectively,x2=0.06,0.02 and 0.01,P=0.800,0.878 and 0.925].Conclusions Pre-implanted anti-adhesion patch under laparoscopic approach can significantly reduce the incidence of parastomal hernia in elderly patients after a Miles' operation,which is worthy of promotion.
5.Moyamoya disease with lenticulostriate artery aneurysm rupture: clinical features, treatment and outcomes
Yi WANG ; Shijie NA ; Tao LIU ; Tianzhu XU ; Jiannan MAO ; Yongbo YANG
International Journal of Cerebrovascular Diseases 2018;26(10):750-755
Objective To investigate the clinical features, treatment strategies and outcomes of patients with Moyamoya disease and lenticulostriate artery (LSA) aneurysm rupture. Methods Patients with moyamoya disease and LSA aneurysm rupture admitted to Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medicine School from October 2012 to March 2015 were analyzed retrospectively. They were followed up for 1 year. The modified Rankin Scale (mRS) was used to evaluate the outcomes, and 0-2 was defined as good outcome. The demographic characteristics, image anatomical features, treatment schemes, and outcomes of the patients were summarized. The Pubmed database was used to retrieve other similar studies, which combined with this group of cases for analysis. Results A total of 10 patients were enrolled, 2 males and 8 females, aged 29-72 years, with an average of 43. 6 years. All cases were intracranial hemorrhage, including 3 cases of cerebral hemorrhage, 6 cases of ventricular hemorrhage, and 1 case of subarachnoid hemorrhage. At the time of admission, 7 patients had disturbance of consciousness, and 3 patients underwent emergency extraventricular drainage. Among them, 5 patients received endovascular embolization, 4 had good outcome, 1 had mild neurological deficit (mRS score 3); 2 received surgical treatment, all had good outcome; 3 received conservative treatment, all had re-bleeding, 2 died, and 1 had severe disability (mRS score 4). Fourteen eligible articles were included, and a total of 18 patients were included in the analysis: surgical treatment in 7 cases, endovascular treatment in 6 cases, and conservative treatment in 5 cases (self-healing in 1 case). According to the data of this group of patients and literature reports, the good outcome rate of the patients with early interventional embolization or craniotomy clipping treatment was significantly higher than that of conservative treatment (70. 6%vs. 22. 2%; P = 0. 038). Conclusion There is a certain risk of conservative treatment of Moyamoya disease complicated with LSA aneurysm rupture and requires active treatment. Interventional embolization of the parent artery or surgically clipping of aneurysm can effectively improve the clinical outcome of such patients. Interventional embolization of the parent artery and aneurysm can be selected simultaneously if the condition of the parent artery is allowed. Surgery can be selected when the condition of parent artery is poor and the aneurysm is located in the superficial part.
6.Biodegradable flow-diverting stent for treatment of saccular wide-neck aneurysm in rabbits
Kuizhong WANG ; Qiang LIU ; Lubin YAN ; Shaoji YUAN ; Zhenqiang HAO ; Tianzhu WANG ; Peigang LU
Chinese Journal of Neuromedicine 2018;17(3):263-268
Objective To evaluate the feasibility of biodegradable flow-diverting stent for treatment of intracranial wide-neck aneurysm.Methods Twenty adult New Zealand rabbits were used to induce wide-neck aneurysm models by elastase,and then,these models were divided into 3 groups:in group one (n=7),biodegradable polyglycolic acid flow-diverting stents (PGA-FDs) were implanted into parent arteries and covered the necks of aneurysms;in group 2 (n=7),the bare metal FDs replaced PGA-FDs;the observation period of animals in group one and group 2 was 3 months;in group 3 (n=6),the PGA-FDs were implanted into parent arteries with 6 weeks of observation period.The status of aneurysm occlusion and patency of side branches covered by the stent were assessed,and all animals were sacrificed:HE staining was used to observe the thrombosis in the aneurysms,Elastic fiber staining was used to observe the neointimal thickness of parent arteries,and immuohistochemical staining was employed to detect the expressions of CD34,CD68 and alpha-actin (αSMA) in the parent arteries.Results Complete aneurysm occlusion rate in groups one,2 and 3 was 83.3% (5/6),0% (0/6) and 66.7% (4/6),respectively.The neointimal thickness in parent artery of groups one,2 and 3 was (111±69) μm,(70±48) μm and (190±52) μm,respectively;that in group 3 was significantly higher than that in group one and 2 (P<0.05).The neointima comprised predominantly smooth muscle cells and collagen fibers.Some CD34 positive vascular endothelial cells could be seen in the neointima of parent arteries of groups one and 2.Conclusion The PGA-FD is an effective device to occlude aneurysms without inducing obvious neointimal thickness and side branch stenosis in the 3-month follow-up.
7.Association of the polymorphisms in TP63 rs6790167 with risk for lung cancer in Hain-an Han population
TANG WENJUN ; YAN QIXING ; WANG TIANZHU ; YI GUOHUI ; XUE LI ; LIN HAIFENG
Chinese Journal of Clinical Oncology 2017;44(21):1061-1066
Objective:To evaluate the association between the polymorphisms in TP63 rs6790167 and lung cancer. Methods:We per-formed a case-control study with 258 patients and 270 controls. Genomic DNA was extracted using a blood genome extraction kit. Ge-notyping was conducted using allele-specific polymerase chain reaction (AS-PCR). Results:No significant difference was observed in the genotype of TP63 rs6790167 between cases and controls, and allele frequencies significantly elevated in the cases (P=0.045). In a further stratified analysis by gender and smoking status, the frequencies of the GG genotype and G allele of rs6790167 were signifi-cantly higher in non-smoking male patients with lung adenocarcinoma (P=0.000, P=0.001), individuals with GG genotype had a higher risk for lung adenocarcinoma in non-smoking man (OR=6.66, 95%CI:2.16-20.41). Individuals with G allele had a higher risk for lung ad-enocarcinoma in a non-smoking man (OR=2.54, 95%CI:1.42-4.56) in comparison with individuals with A allele. Conclusion:We found that the polymorphisms of TP63 rs6790167 were associated with the risk for lung adenocarcinoma in non-smoking men and that these polymorphisms may be a predictor for lung adenocarcinoma in non-smoking men.
8.Purification Method of Ultrasonic Emulsification Combined with Ultrafiltration Membrane Separation for Flavonoids in Cacumen Platycladi
Xianyi WANG ; Ruihai LI ; Tianzhu JIA
China Pharmacist 2017;20(4):661-663,667
Objective:To obtain flavonoids of Cacumen Platycladi with high purity by using the method of ultrasonic emulsification combined with ultrafiltration membrane separation.Methods:Seven compounds including myricetrin as the reference substances,an HPLC method was used for the content determination.Using the single factor experiments,ultrasound,microwave,reflux and ultrasonic emulsification were compared,and using the orthogonal experiments,the ultrasonic emulsification time,ethanol concentration,solid-liquid ratio and times were studied.Biomax-5 membrane was adopted to improve the membrane separation.Results:The single factor experiments showed the homogeneous extraction method with the highest contents.The orthogonal experiments showed the optimal extracting conditions as follows:the ultrasonic emulsification time was 15 min,the ethanol concentration was 50%,the material-liquid ratio was 1∶10,and the extraction times was 3.The conditions of ultrafiltration membrane separation were as follows:the flow rate was 1.5 L·min-1,and the membrane separation pressure was 0.8 kg.Conclusion:The combination of ultrasonic emulsification and ultrafiltration membrane separation is feasible in the extraction and purification of flavonoids in Cacumen Platycladi,and the product is with high purity,suggesting the method has good application prospect.
9.Identification of targeting relationship between mouse lncRNA-H19 and miR-199 a-5 p by dual-luciferase reporter assay
Jingying HOU ; Changqing ZHOU ; Shaoxin ZHENG ; Tianzhu GUO ; Huibao LONG ; Quanhua WU ; Tingting ZHONG ; Hao WU ; Lei WANG ; Tong WANG
Chinese Journal of Pathophysiology 2016;32(12):2256-2260
AIM:To validate the association between long noncoding (lncRNA)-H19 and microRNA-199a-5p (miR-199a-5p) through the dual-luciferase reporter gene system by construction of a luciferase reporter vector containing the gene of lncRNA-H19.METHODS:The potential complementary binding sites of lncRNA-H19 and miR-199a-5p were predicted by RegRNA 2.0.The H19 gene or its mutant ( Mut) fragment was cloned into luciferase reporter vector psi-CHECK-2.Restriction enzyme analysis and sequence analysis were used to identify whether the recombinant plasmids of the H19 and H19-Mut were successfully constructed .miR-199a-5p mimics, miR-199a-5p inhibitor, miR-199a-5p mimics neg-ative control or miR-199a-5p inhibitor negative control was co-transfected into the 293T cells with the luciferase reporters containing H19 or H19-Mut.Dual-luciferase reporter assay was performed to detect the luciferase activity in different groups in order to verify the relationship between lncRNA-H19 and miR-199a-5p.RESULTS:The results of double enzyme diges-tion and DNA sequencing showed that the sequence of luciferase reporter vector was correct .The results of dual-luciferase reporter assay indicated that the H 19 reporter gene luciferase activity significantly decreased in miR-199a-5p mimics group by 49%(P<0.01), and the H19 reporter gene luciferase activity was obviously upregulated in miR-199a-5p inhibitor group compared with miR-199a-5p mimics group ( P<0.01).However, miR-199a-5p mimics, miR-199a-5p inhibitor, miR-199a-5p mimics negative control and miR-199a-5p inhibitor negative control showed no effect at H 19-Mut reporter gene.CONCLUSION:lncRNA-H19 binds to miR-199a-5p to exert an inhibitory effect at transcriptional level .
10.Research progress of alternative in vitro methods to evaluate skin sensitization
Ning CHEN ; Ping WANG ; Jingwen XIAN ; Tianzhu XIE ; Meirong QIN ; Junpeng LI ; Yi LU ; Xiaowei WANG ; Haining SHI ; Shaorong LONG
Chinese Journal of Comparative Medicine 2016;26(12):85-90
Skin sensitization (allergic contact dermatitis, ACD), is a serious condition caused by small reactive molecules and is characterized by a delayed-type hypersensitivity .Animal tests were usually used in the evaluation of sensitizing potential of chemical substances in the past .However , there is an increasing interest from the public for reducing and ultimately replacing animal tests .The European Union (EU) has posed a ban on animal testing of cosmetic ingredients that includes skin sensitization since 2013.Therefore, alternative in vitro tests are the main tendency in chemical substances and cosmetic sensitizing potential research in the future .In this study, different kinds of in vitro test methods that were adopted by OECD or on research (LLNA, DPRA, KeratinoSens TM, h-CLAT) were reviewed through recent years literature , comprehensive introduction and evaluation were made to obtain reliable hazard and potency information on potential skin sensitizers .

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