1.Analysis of microbiota characteristics in tumor tissues sampled under gastroscope
Yingchun LI ; Yue YIN ; Dalin LU ; Huan XU ; Fang WANG ; Yanbo ZHENG ; Zhaoming JIA ; Tao SHEN ; Zhinong SHI
Chinese Journal of Microbiology and Immunology 2022;42(7):562-569
		                        		
		                        			
		                        			Objective:To investigate the characteristics of bacterial community in upper gastrointestinal tumors.Methods:The study population was patients with upper gastrointestinal tumors (esophageal cancer and gastric cancer). Gastroscopy was performed on the enrolled patients ( n=17), and the specimens were taken from the tumor sites. At the same time, non-tumor tissues more than 4 cm away from the tumor tissues were taken as the control. After total DNA was extracted and purified, high-throughput 16S DNA gene sequencing was used to detect the microbiota in tumor tissues and control tissues. Bioinformatics analysis was carried out and the differences between groups were compared. Results:16S DNA PCR showed that there was no significant difference in bacterial load between tumor tissues and control tissues. The α-diversity and β-diversity indexes showed that the community composition of the two groups was similar; the samples were discrete and the colony composition was different, but there was no significant difference between the two groups. The results of Venn diagram showed that there were more operational taxonomic units (OTUs) in non-tumor tissues than in tumor tissues (2 068 vs 1 358), indicating that the bacterial species in normal tissues were more abundant than those in tumor tissues. Compared with the control tissues, the percentages of Prevotellaceae ( Prevotella), Lactobacaceae ( Lactobacillus) and Fusobacteriaceae ( Fusobacterium) in tumor tissues were relatively higher (the average percentage was more than twice that of the control). Further paired comparison of the top ten bacteria in the family and genus abundance of the two groups of samples showed that Pseudomonas decreased significantly in tumor tissues at the family ( P=0.041) and genus ( P=0.041) levels, while Prevotella was significantly enriched in tumor tissues at the family ( P=0.031) and genus ( P=0.007) levels. Conclusions:The bacterial community in the tumor microenvironment of the upper gastrointestinal tumor changed, and the species enriched in the tumor site were mainly oral common anaerobic bacteria, such as Prevotellaceae ( Prevotella), Lactobacaceae ( Lactobacillus) and Fusobacteriaceae ( Fusobacterium), especially Prevotellaceae ( Prevotella).
		                        		
		                        		
		                        		
		                        	
2.Clinical characteristics and complications after vitrectomy in patients with vitreous amyloidosis from three Han nationality families
Yanbing FENG ; Wenqing WENG ; Yanyan HE ; Zhenyi MA ; Yanbo SHI ; Yibo WU ; Yixing ZHU ; Zhixin SHEN
Chinese Journal of Ocular Fundus Diseases 2021;37(11):865-871
		                        		
		                        			
		                        			Objective:To observe the clinical characteristics of patients with familial vitreous amyloidosis (FVA) and the efficacy of vitrectomy (PPV) and the occurrence of complications.Methods:A retrospective clinical study. From June 2009 to March 2020, 32 eyes of 18 patients from 3 FVA families who were diagnosed and treated by PPV at Department of Ophthalmology of Jiaxing TCM Hospital were included in the study. Among them, there were 12 males with 22 eyes and 6 females with 10 eyes. The average age of onset was 42.28±3.25 years; the average duration of disease was 3.75±3.93 years. All the affected eyes underwent best corrected visual acuity (BCVA) and B-mode ultrasound examination. A logarithmic visual acuity chart was used in the BCVA examination, which was converted to the logarithmic minimum angle of resolution (logMAR) visual acuity when recorded. The average logMAR BCVA of the affected eye was 1.72±0.53; the intraocular pressure was less than 21 mm Hg (1 mm Hg=0.133 kPa). The vitreous body of the affected eye was obviously cloudy. All the affected eyes underwent standard three-channel PPV through the flat part of the ciliary body, and vitreous specimens were collected for pathological examination during the operation. Peripheral venous blood of probands from 3 families was collected, and the whole exome gene sequencing was performed. The follow-up time after surgery was ≥6 months. The patient's clinical characteristics, fundus lesions in PPV, changes in BCVA after surgery, and complications was observed. One-way analysis of variance or t test was performed for measurement data comparison; χ2 test was performed for count data comparison. Results:The vitreous body of the affected eye showed gray-white dense and thick flocculent changes, and the posterior capsule attached to the lens showed "foot disc-like" turbidity; later the lens was mainly cystic opacity. Pathological examination of the vitreous body showed positive staining of Congo red; under a polarized light microscope, it showed apple green dots and sheet-like birefringence. The genetic test results showed that there was a c.307G>C (p.Gly103Arg) missense mutation in the TTR gene of the proband in Family 2. Peripheral retinal hemorrhages in 4 eyes (12.5%, 4/32), retinal tears in 5 eyes (15.6%, 5/32), retinal degeneration in 4 eyes (12.5%, 4/32), retinal detachment were found in PPV 3 eyes (9.4%, 3/32). The vitreous body was filled with C 3F 8 and silicone oil respectively for 2, 1 eye. Six months after the operation, the logMAR BCVA of the affected eye was 0.39±0.32, which was significantly higher than that before the operation, and the difference was statistically significant ( t=15.131, P=0.000). After the operation, high intraocular pressure occurred in 2 eyes (6.3%, 2/32), secondary glaucoma in 1 eye (3.1%, 1/32), retinal detachment in 2 eyes (6.3%, 2/32), neovascular glaucoma (NVG) in 2 eyes (6.3%, 2/32), cataract in 10 eyes (31.3%, 10/32). Conclusion:The vitreous body of FVA eyes are gray-white dense, thick and flocculent, attached to the posterior lens capsule, showing "foot disc-like" turbidity; PPV treatment can effectively improve the BCVA of the FVA eyes; secondary glaucoma, secondary retinal detachment, NVG can occur after surgery.
		                        		
		                        		
		                        		
		                        	
3.miR-423-5p enhances chemoresistance of glioblastomas cells to temozolomide by regulating PDCD5
WANG Lei ; SHEN Weigao ; LIU Yanbo
Chinese Journal of Cancer Biotherapy 2020;27(10):1100-1105
		                        		
		                        			
		                        			[Abstract] Objective: To investigate the miR-423-5p expression in brain glioma tissues and cell lines, and its promotive effect on
temozolomide (TMZ) chemoresistance by targeting PDCD5 (programmed cell death protein 5). Methods: Tumor tissues and matched
peritumoral tissues were collected from 20 brain glioma patients who were surgically treated in the Department of Neurosurgery,
Affiliated Hospital of Beihua University between January 2017 and December 2018. Glioblastoma cell lines (U251, U87, SHG-44)
and human normal glial cell line HMC-3 were also used in the study. The relative expression of miR-423-5p and PDCD5 in brain
glioma and peritumoral tissues and cell lines was detected by qPCR. The synthesized miR-423-5p mimics and miR-NC were respectively
transfected into U251 and U87 cells; meanwhile, TMZ at different concentrations (50, 100, 150 and 200 μmol/L) were also used to treat
the cells. Then, the chemoresistance of cells to TMZ were determined. MTT assay and colony formation assay were used to examine the
proliferation of U251 and U87 cells, andWestern blotting was used to detect the expression of c-caspase 3, Bcl-2 and PDCD5 proteins in
U251 and U87 cells. The targeting relationship between PDCD5 and miR-423-5p was validated through Dual luciferase reporter gene
assay. Results: miR-423-5p was highly expressed in glioma tissues and glioma cell lines (all P<0.01). As compared with the miR-NC
group, the proliferation and TMZ-chemoresistance of U251 and U87 cells in miR-423-5p mimics group significantly increased (all
P<0.01). Dual luciferase reporter gene assay validated that miR-423-5p could bind with PDCD5 3' UTR to suppress the expression of
PDCD5. Conclusion: High expression of miR-423-5p enhances the chemoresistance of glioma cells to TMZ, and miR-423-5p may
serve as a potential therapeutic target in the treatment of brain glioma.
		                        		
		                        		
		                        		
		                        	
4. Berberine alleviates cerebral ischemia-reperfusion injury in type 2 diabetic rats by inhibiting endoplasmic reticulum stress-related inflammatory pathways
Chong LIU ; Xu YANG ; Na SHEN ; Yanbo DI ; Xuan LIU ; Qiang CHEN ; Feng HE ; Huanming LI
Chinese Journal of Geriatrics 2020;39(1):73-77
		                        		
		                        			 Objective:
		                        			To examine the effects of Berberine(BBR)on inflammatory pathways related to endoplasmic reticulum stress(ERS)in the penumbra area following focal cerebral ischemia-reperfusion injury in type 2 diabetic rats.
		                        		
		                        			Methods:
		                        			A total of 72 healthy male Sprague-Dawley rats were fed a high-fat, high-sugar diet and injected with streptozotocin to establish a type 2 diabetes mellitus model.The diabetic rats were randomly divided by digital lottery method into a Sham operation group(Sham group), a diabetic rat + BBR treatment group(BBR group), a diabetic middle cerebral artery occlusion(MCAO)model group(MCAO group), and a diabetic rats MCAO + BBR treatment group(MCAO + BBR group). Six rats were included in each group.The two treatment groups received prespecified doses of BBR through gastric perfusion at 48 h, 24 h before surgery, and 6h after surgery.The MCAO model was prepared by a suture occlusion method.The neurological deficit scores were performed, and the expression of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)was detected by enzyme-linked immunosorbent assay(ELISA). The mRNA expression of ERS marker protein GRP78 was detected by quantitative real time polymerase chain reaction(RT-qPCR), and the expression of ERS-related inflammatory pathway proteins[78 Glucoseregulated protein(GRP78)、Pancreatic endoplasmic reticulum Rinase(PERK)、nuclear factor-κB(NF-κB)]was detected by Western blot.
		                        		
		                        			Results:
		                        			the cerebral ischemic penumbra area, after 2 h of ischemia and 24 h of reperfusion, the neurological deficit score in the MCAO group was higher than that in the MACO+ BBR group [(2.83±0.41)
		                        		
		                        	
5.Over-expression of programmed cell death 5 gene enhances chemosensitivity of brain glioma cells to temozolomide
WANG Lei ; SHEN Weigao ; LIU Yanbo ; XU Haiyang
Chinese Journal of Cancer Biotherapy 2019;26(8):868-875
		                        		
		                        			
		                        			 Objective: To explore the role of tumor suppressor gene programmed cell death 5 gene (PDCD5) in the growth and temozolomide (TMZ) sensitivity of brain glioma cells. Methods:Atotal of 116 patients with cerebral glioma admitted to the Department of Neurosurgery, First Clinical Hospital of Jilin University from January 2009 to December 2014 were enrolled in this study. QPCR, WB and immunohistochemistry method were used to detect the mRNAand protein expressions of PDCD5 in glioma cell lines (U87, U251), U87 cell line with stable PDCD5 expression (U87-PDCD5), glioma cells with si-PDCD5 transfection and primary cerebral glioma tissues, respectively. MTT assay was used to detect the effect of over-expression or knockdown of PDCD5 on the growth and TMZ-sensitivity of glioma cells. The subcutaneous tumor-bearing model of glioma cell line U87 was established in nude mice, and then the experimental mice were randomly divided into control group, TMZ group, PDCD5 group and TMZ+exogenous PDCD5 recombinant expression vector group.After 20 days, the animals were sacrificed by cervical dislocation and the tumor tissue was excised to measure the tumor volume and weigh. The expression of PDCD5 in tumor tissues was detected by qPCR and WB methods, and the effects of PDCD5 combined with TMZ on the growth of gliomas were also analyzed. Results: The relative mRNA and protein expressions of PDCD5 in U87 cells were significantly lower than those in U251 cells (both P<0.05), and the mRNA and protein expressions of PDCD5 in high level glioma tissues were significantly lower than those in low level tissues (all P<0.05). The sensitivity of U87-PDCD5 cells and U251 cells to TMZ was higher than that of U87 cells (all P<0.05). The sensitivity of cells to TMZ in U87-PDCD5-siRNA group and U251siRNA group was significantly lower than that of the control group (both P<0.05). The tumor volume and weigh to fnudemicexenografts were compared,and the results showed control group>TMZ group>PDCD 5group>combined group(allP<0.05);however, the mRNA and protein expressions of PDCD5 in the transplanted tumor tissues of each group showed the opposite trend (all P<0.05). Conclusion: PDCD5 over-expression can enhance the chemosensitivity of braingliomato the chemotherapy drug TMZ, while silencing of PDCD5 expression exertsthe opposite effect.The combination of PDCD5 and TMZ can better inhibit the growth of xenografts in nude mice. 
		                        		
		                        		
		                        		
		                        	
6.Effect of Berberine on endoplasmic reticulum stress and insulin resistance in skeletal muscle of metabolic syndrome rats and its mechanism
Hong ZHANG ; Tao FANG ; Fengshi TIAN ; Yanbo DI ; Na SHEN ; Ying CHEN ; Qiang CHEN ; Feng HE ; Yang BAI ; Huanming LI
Chinese Journal of Geriatrics 2019;38(8):924-928
		                        		
		                        			
		                        			Objective To investigate the effect of Berberine on insulin resistance and its mechanism in Otsuka Long-Evans Tokushima Fatty(OLETF) rats with metabolic syndrome(MS).Methods LETO(Long-evans Tokushima Otsuka)rats(the control group receiving standard normal diet,n=10)and diabetic OLETF rats(the MS group receiving high-fat diet for 24 weeks,n=30).Rats in the MS group were randomly divided into 3 subgroups(n=10,each subgroup).Each subgroup was gavaged with normal saline,high-dose Berberine(100 mg · kg-1 · d-1)and low-dose Berberine (50 mg · kg-1 · d-1) respectively,and the high-fat diet remained unchanged.After 6 weeks of berberine treatment,body weight,blood glucose and lipid metabolism parameters were determined.The oral glucose tolerance test(OGTT) and insulin tolerance test(ITT) were used to detect insulin resistance.Expression levels of the protein and mRNA of 78 kDa glucose-regulated protein (GRP7 8),Caspase-12 and CCAAT/enhancer-binding protein(C/EBP) homologous protein(CHOP) in skeletal muscles were detected by Western blot and RT-PCR.Results After Berberine treatment,the body weight,fasting plasma glucose,fasting insulin[(28.9 ± 2.0) mU/L,(31.5± 2.4) mU/L vs.(36.9 ± 4.7) mU/L],total cholesterol,triglycerides,and low-density lipoprotein cholesterol were decreased,while the high-density lipoprotein cholesterol(HDL-C) levels were increased in MS rats with high-dose berberine and low-dose berberine as compared with the control group (P < 0.05) respectively.Berberine treatment could reduce the protein and mRNA expression levels of GRP78,Capase-12 and CHOP in the skeletal muscle of MS rats(P<0.05).Conclusions Berberine may alleviate insulin resistance in rats with metabolic syndrome by reducing endoplasmic reticulum stress in skeletal muscle.
		                        		
		                        		
		                        		
		                        	
7.Clinical features of candidemia and the diagnostic value of (1, 3) -β-D glucan
Wang SHEN ; Wenli YANG ; Yanbo CHEN ; Yuegui LI ; Yimei ZHONG ; Xin ZHANG
Chinese Journal of Infection and Chemotherapy 2019;19(1):42-47
		                        		
		                        			
		                        			Objective To understand the etiology and clinical characteristics of candidemia, and the diagnostic value of(1, 3)-β-Dglucan. Methods This analysis included 31 patients with candidemia, 39 patients with Candida deep colonization, and 50 healthy subjects, who were enrolled from Wuyi Chinese Hospital Affiliated to Jinan University during the period from September 2015 to March 2018. Clinical data such as fungal culture, clinical features, and serum(1, 3)-β-D-glucan level were analyzed and compared between groups. Results Overall, 13 strains of Candida albicans and 18 strains of non-albicans Candida were isolated from the 31 patients with candidemia. The incidence of candidemia was associated with old age(>65 years), prior surgery(≤7 days), indwelling catheter(≥2 catheters and lasting ≥5 days), duration of mechanical ventilation(≥5 days)(P<0.05), but not associated with sex, underlying disease, prior use of anmicrobial and antifungal agents, immunosuppressant or glucocorticoid, or hypoproteinemia(P>0.05). Serum level of(1, 3)-β-D-glucan was significantly higher in candidemia patients than in the patients with candida deep colonization and healthy controls(P<0.05). The sensitivity and specificity of(1, 3)-β-D-glucan in diagnosing candidemia were 87.1% and 96.0%, respectively. The level of(1, 3)-β-D-glucan was significantly higher in candidemia with bacterial infection than candidemia without bacterial infection(P<0.05). The level of of(1, 3)-β-D-glucan was also significantly higher in the candidemia caused by Candida albicans alone than that caused by non-albicans Candida alone(P<0.05). Conclusions Non-albicans Candida is more prevalent than Candida albicans in candidiasis, but Candida albicans is still the most important Candida species. The incidence of candidemia is asociated with advanced age(>65 years), prior surgery within 7 days, indwelling catheters(≥2 catheters and lasting ≥5 days), and mechanical ventilation(≥5 days). Serum(1, 3)-β-D-glucan level is valuable for early diagnosis of candidemia.
		                        		
		                        		
		                        		
		                        	
8.The exploration of curriculum system of humanistic education with the whole process and multidis-ciplinary integration for medical students
Yanbo WANG ; Xudong ZHAO ; Qiaoling CAI ; Huixia SHEN ; Jingyu SHI ; Hao ZHENG ; Zhaoxin WANG
Chinese Journal of Medical Education Research 2016;15(2):157-160
		                        		
		                        			
		                        			Based on the analyses of the status of humanistic medicine education both at home and abroad, the article emphasized the need for the implement of medical humanities education with the whole process and multidisciplinary integration. In combination with practice, from the three modules of the ex-plicit curriculum, implicit curriculum and integrated curriculum, the author discussed the specific conno-tation of the whole course of medical humanities education. The article also summarized the main points of the course system in teaching practice from the aspects of training objectives, teaching staff construction, teaching methods improvement and innovation, and humanistic quality evaluation of medical students.
		                        		
		                        		
		                        		
		                        	
9.A study on the expressions of IL-17A and IL-17RA in different degrees of malignant glioma
Lei WANG ; Yanbo LIU ; Zhenjiang WANG ; Xinli ZHAO ; Weigao SHEN
Tianjin Medical Journal 2016;44(8):1018-1021
		                        		
		                        			
		                        			Objective To explore the expressions of interleukin (IL)-17A and its receptor IL-17RA in different degrees of malignant gliomas. Methods Fifty patients with glioma were collected in this study. Accordance to the World Health Organization Classification System, patients were classified by malignancy grade, including gradeⅠ(n=12), gradeⅡ(n=18), gradeⅢ(n=13) and gradeⅣ(n=7). The glioma tissue and peripheral blood samples of patients were obtained for detecting the expression levels of IL-17A and IL-17RA mRNA by using immunohistochemistry, quantitative real-time PCR. Western blot assay was used to detect expressions of IL- 17A and IL- 17RA in both the macroscopic (immunohistochemistry) and molecular levels (mRNA and protein). Results Immunohistochemical staining showed that the expression levels of IL-17A and its receptor IL-17RA increased with the increase of the malignant degree of gliomas. The mRNA levels of IL-17A and IL-17RA receptors in peripheral blood were up-regulated with the increasing malignancy grade of glioma (F=8.96, P<0.05;F=10.34, P<0.05). The mRNA levels of IL-17A and IL-17RA in glioma tissues were up-regulated with the increasing malignancy grade of glioma (F=11.21, P<0.05;F=14.11, P<0.05). The protein levels of IL-17A and IL-17RA in peripheral blood and glioma tissues were also up-regulated with the increasing malignancy grade of glioma (in peripheral blood:F=9.90, P<0.05;F=11.80, P<0.05;and in gliomas tissues:F=8.15, P<0.05;F=14.46, P<0.05). Conclusion The expressions of IL-17A and IL-17RA receptor are positively correlated with malignancy grade of glioma. These results provide some reference for clinical diagnosis of malignant gliomas.
		                        		
		                        		
		                        		
		                        	
10.Comparison clinical curative effect of Sanxuemingmu tablets and compound Xueshuantong capsulein treatment of RVO patients with qi stagnation and blood stasis syndrome
Chinese Journal of Biochemical Pharmaceutics 2016;36(12):115-117
		                        		
		                        			
		                        			Objective To investigate the efficacy and mechanism of Sanxuemingmu tablets and compound Xueshuantong capsule in treatment of RVO patients with qi stagnation and blood stasis syndrome.Methods 1200 cases of RVO patients with qi stagnation and blood stasis syndrome from April 2014 to April 2016 months in our hospital were selected,and randomly divided into two groups,600 cases in each group.The observation group was treated with Sanxuemingmu tablets,the control group were treated with compound Xueshuantong capsule,the curative effect and relative score of the two groups were compared.Results After treatment,the visual acuity of the two groups was significantly higher than that before treatment,the visual acuity of the observation group was higher than control group (P<0.05);The absorption of fundus hemorrhage in the observation group was significantly better than that in the control group(P<0.05);The total effective rate of FFA in observation group was significantly higher than that in control group (P<0.05);Compared with before treatment,the whole blood viscosity and plasma viscosity of the two groups were decreased,and the whole blood viscosity and plasma viscosity of the observation group were lower than those of the control group(P<0.05);The TCM syndrome scores of both groups were lower than those before treatment,and the observation group was significantly lower than the control group( P <0.05);The total effective rate and the total effective rate of the treatment group were significantly higher in the observation group than in the control group(P<0.05).Conclusion Sanxuemingmu tablets has significant clinical effect in treatment of RVO patients with qi stagnation and blood stasis syndrome,can accelerate the absorption of retinal hemorrhage,treatment of retinal vein obstruction qi stagnation of blood stasis,promote vision recovery and improve treatment efficiency.
		                        		
		                        		
		                        		
		                        	
            
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