1.Effect of Faecalibacterium prausnitzii in Colitis-associated Colorectal Cancer
Lijuan XIE ; Xuejia LU ; Chenggong YU
Chinese Journal of Gastroenterology 2015;(9):517-522
Background:Faecalibacterium prausnitzii( Fp) is one of the most abundant bacterium in human intestinal microbiota,and is closely correlated with the process of colitis-associated colorectal cancer(CAC). Aims:To observe the effect of Fp on CAC,and investigate the possible mechanism. Methods:The model of CAC was induced by azoxymethane (AOM)and dextran sodium sulfate( DSS). Fifty-two C57BL/ 6J mice were randomly divided into 4 groups:group A (AOM + DSS),group B(AOM + DSS + Fp),group C(AOM + DSS + Fp supernatant)and group D(control group). All the mice were sacrificed on day 92. DAI was assessed,serum levels of TNF-α and IL-10 were determined by ELISA. HE staining was used to examine the grade of tumor. Expressions of VEGF,COX-2,NF-κB in tumor tissue were measured by immunohistochemistry. Results:The tumorigenesis rates of group A,B,C were 100% ,100% and 77. 8% ,respectively;mainly were high-grade intraepithelial neoplasia. The tumor load in group A was significantly higher than that in group B (P < 0. 01),and the spleen index in group B was significantly higher than that in group C(P < 0. 01). Serum level of TNF-α was significantly lower(P < 0. 05)and IL-10 was significantly higher(P < 0. 05)in group A than that in group B. No significant differences in expressions of VEGF,COX-2,NF-κB were found among group A,B and C. Conclusions:Fp had no obvious effect on the occurrence rate of CAC,and Fp supernatant could decrease the incidence of CAC in mice. Fp and its supernatant could reduce the tumor load via regulating the expressions of TNF-α,IL-10.
2.Process of Purification of Dihydromyricetin by Cooling Crystallization
Xuejia XIE ; Chenguang WANG ; Xiaolong HOU ; Wei XIONG ; Wenqing WANG
Herald of Medicine 2017;36(8):912-917
Objective To development a cooling crystallization process that is suitable for industrial preparation of purified dihydromyricetin.Methods Screen design was used to investigate effects of process parameters such as,temperature,concentration ethanol aqueous,quantity of activated charcoal and adsorption time on yield and purity of dihydromyricetin.Purity was verified by high performance liquid chromatography and thin layer chromatography.The solubility of dihydromyricetin in water at viable temperature and ethanol proportion was also determined by UV spectrophotometry.The solid form was characterized by thermal analysis and powder X-ray diffractometry.Results When temperature was > 85 ℃,ethanol concentration < 10%,dosage of activated charcoal 0.1%-0.3%,and adsorption time 1-3 min,yield of dihydromyricetin was more than 70%,and the purity greater than 98%.The crystals,prepared by cooling crystallization from water and ethanol aqueous,had the same physical form and crystal habit.Conclusion Cooling crystallization from low concentration of ethanol aqueous gets higher yield and the process is more robust than crystallization from water.
3.Clinical observation of formula for clearing heat,eliminating phlegm and relaxing bowel on treating encephalopathy complicated with pulmonary infection
Junying LV ; Kai LI ; Liping HUANG ; Aize XIE ; Xuejia ZHENG ; Juan LIANG ; Shuangyan LIU
The Journal of Practical Medicine 2017;33(6):1001-1004
Objective The efficacy and safety of rectal instillation with formula for clearing heat, eliminating phlegm and relaxing bowel on treating encephalopathy complicated with pulmonary infection were evaluated by randomized controlled clinical trials. Methods Eighty eligible cases of encephalopathy complicated with pulmonary infection were randomly divided into treatment group and control group. Control group was given conventional treatment and treatment group was given rectal instillation treatment with formula for clearing heat , eliminating phlegm and relaxing bowel. The changes of index were observed before and after treatment in two groups including temperature,antifebrile time,white blood cell count,neutrophils ratio,C-reactive protein,procalcitonin, pneumonia severity index,and the efficacy score of TCM syndrome. Results Rectal instillation with formula for clearing heat and eliminating phlegm to relax bowel can be better improved encephalopathy complicated with pulmonary infection with syndrome of phlegm-heat obstructing in which the patients have yellow sputum, constipation,yellow urinary and other symptoms. It can reduce TCM syndrome score and level of serum calcitonin, relieve inflammation,enhanc antibacterial effect,and improve the clinical curative effect. Conclusions The curative effects of rectal instillation with formula for clearing heat and eliminating phlegm to relax bowel on treating encephalopathy complicated with pulmonary is definite which has no adverse reaction and safety use. It is worthy of further popularizing in clinic.
4.Effect of FOXP3 expression in pancreatic carcinoma cells on the maturation and immunologic function of dendritic cells
Xuejia LU ; Ying LYU ; Chao MA ; Wenjia LIU ; Lijuan XIE ; Xiaoping ZOU
Chinese Journal of Pancreatology 2015;15(5):325-330
Objective To investigate the influence of FOXP 3 expression in pancreatic carcinoma cells (PCCs) on the maturation and immunologic function of dendritic cells (DCs).Methods The siRNA sequences targeting FOXP3 gene (siRNA-FOXP3) and negative control siRNA (siRNA-NC) were specifically designed and transfected into PCCs , then the level of IL-10 and TGF-β1 of culture supernatant were detected by ELISA.The supernatants of pancreatic carcinoma cell transfected by FOXP 3-siRNA were collected , then it was mixed with GM-CSF and IL-4 to induce the differentiation of DCs .Flow cytometric analysis were used to measure the expression of surface markers CD 86 , CD80 , HLA DR on DCs which were treated with supernatants . The levels of IL-12p70, IFN-γin supernatants were detected by ELISA .The DCs were co-cultured with T lymphocytes, and then the lymphocytes proliferation and cytoxicity were analyzed by CCK -8 assays.Results Compared with PANC1 with siRNA-NC transfection, PANC1 with siRNA-FOXP3 transfection had a decreased expression of IL-10, TGF-β1 [(8.93 ±3.06)ng/L vs (26.60 ±5.57)ng/L;(2 544 ±78)ng/L vs (2 856 ± 92)ng/L], the positive expression rate of CD86, HLA DR in DCs cultured in the medium containing the supernatants of the pancreatic carcinoma cell transfected by siRNA-FOXP3 was significantly increased [(28.10 ±3.11)%vs (13.90 ±0.42)%;(66.15 ±4.17)%vs (43.15 ±3.32)%], the expression of IL-12p70, IFN-γwas significantly increased [(52.75 ±7.89)ng/L vs (26.14 ±4.50)ng/L, (898.43 ±88.82) ng/L vs (412.76 ±24.68) ng/L], after co-culture with lymphocytes at ratios of 1:5, 1:10, 1:20, the proliferation was significantly increased [(95.27 ±3.80)% vs (71.77 ±5.70)%, (78.97 ±5.73)% vs (52.30 ±8.72)%, (57.60 ±4.36)% vs (43.73 ±6.01)%], and the cytoxicity of CTL to PANC1 cells with 1:20, 1:40 co-culture was significantly increased [(28.44 ±5.20)% vs (8.82 ±2.29)%, (40.85 ± 5.15)% vs (17.38 ±4.86)%], and the difference between the two groups was statistically significant (P<0.05).Conclusions FOXP3 expression in PCCs can inhibit the maturation and immunologic function of DCs.
5.Diagnostic value of endoscopic ultrasonography for small common bile duct stones
Xuejia LU ; Ting YU ; Ting XIE ; Ruihua SHI
Chinese Journal of Digestive Endoscopy 2022;39(12):1018-1021
To evaluate the diagnostic value of endoscopic ultrasonography (EUS) for small common bile duct stones, 60 patients diagnosed as having suspected common bile duct stones who were hospitalized in Zhongda Hospital, Southeast University from January 2018 to July 2021 were selected. All patients were examined by EUS and magnetic resonance cholangiopancreatography (MRCP) during the same hospitalization. The results of endoscopic retrograde cholangiopancreatography, laparotomy or laparoscopic common bile duct exploration were adopted as the golden standard. The diagnostic results of EUS and MRCP were compared with the golden standard, and the sensitivity, specificity, positive predictive value and negative predictive value of the two methods were calculated. Among the 60 patients, 46 cases were confirmed as having common bile duct stones, 43 cases were diagnosed accurately by EUS and 35 cases by MRCP. Fourteen patients were confirmed as having no common bile duct stones, and the diagnosis was accurate in 12 cases by EUS and 13 cases by MRCP. The sensitivity of EUS was significantly higher than that of MRCP [93.48% (43/46) VS 76.09% (35/46), χ2=4.128, P=0.042]. There were 45 cases with diameter ≤1.0 cm, of which 42 cases were diagnosed by EUS and 34 by MRCP (diagnostic accuracy 93.33% VS 75.56%, χ2=4.145, P=0.042). There were 39 cases with diameter ≤0.8 cm, of which 36 cases were diagnosed by EUS and 28 by MRCP (diagnostic accuracy 92.31% VS 71.79%, χ2=4.266, P=0.039). There were 26 cases with diameter ≤0.5 cm, of which 24 cases were diagnosed by EUS and 16 by MRCP (diagnostic accuracy 92.31% VS 61.54%, χ2=5.038, P=0.021). EUS has obvious advantages in the diagnosis of common bile duct stones, and the accuracy of EUS is not affected by the size of stone. As a consequence, EUS should be performed in patients with high clinical suspicion of common bile duct stones but negative MRCP result.
6.Determination of Related Substances in Lysine Hydrochloride and Zinc Gluconate Oral Solution by High Performance Liquid Chromatography
Xuejia XIE ; Jingyan RAN ; Wenqing WANG
Herald of Medicine 2017;36(12):1394-1398
Objective To establish a HPLC method for the determination of related substances in lysine hydrochloride and zinc gluconate oral solution. Methods The HPLC separation was performed at 30 ℃ on an Inertsil Amide column (4.6 mm×250 mm,5 μm) by gradient elution with mobile phase A [phosphatesolution (2.72 g KH2PO4and 0.46 goctane sulfonate,added with 1 000 mL)-acetonitrile (22:78)] and B [phosphate solution-acetonitrile (95:5)] at the flow rate of 1.0 mL·min-1.The clumn temperature was 30 ℃.The determination wavelength was 203 nm. Results The major peaks were well separated from adjacent related substances peaks and resolution were greater than 2.0.The theoretical plate number of major peaks and related substance peaks were greater than 3 000.Excipients and destruction test products did not interfere with the determination. Conclusion This method is practicable,specific and durable for the simultaneous determination of related substances in lysine hydrochloride and zinc gluconate oral solution.