3.Detection of HIV nucleic acid and subtype analysis in 15 intravenous drug abusers
Nanping WU ; Kezhou LIU ; Zhi CHEN
Chinese Journal of Infectious Diseases 1999;0(01):-
Objective To investigate the molecular epidemiology as well as the subtypes of HIV in intravenous drug abusers, which can help pursue the source of infection in Zhejiang province and predict the epidemic strain in the future. Methods Such as ELISA, Western blot, nest PCR, DNA and sequencing technologies were used to analyze HIV subtypes of 15 strains isolated from intravenous drug abusers(IVDU) from Xinjiang autonomous region. Results All of the 15 IVDUs were infected with HIV 1, 2 of which were infected by subtype E and the others were infected by subtype C. Conclusions Subtype C is main HIV subtype infecting IVDUs. The epidemic subtype may have changed in China.
4.CdTe quantum dot inhibits cell survival and induces mitochondrial dysfunction in primary human umbilical vein endothelial cells
Ming YAN ; Yun ZHANG ; Kezhou LIU ; Yonghong SUN
Chinese Journal of Pharmacology and Toxicology 2017;31(6):574-580
OBJECTIVE To elucidate the toxicological properties of CdTe quantum dots (CdTe QD) by investigating their effect on mitophagy in human umbilical vein endothelial cells (HUVECs).METHODS The purity of primarily cultured HUVECs was detected by immunofluorescence.Then,HUVECs were incubated with CdTe QD 0.1-100 mg· L-1 for 24 h.After treatment,the cell viability of HUVECs was detected with MT-T assay.The mitochondrial morphology was observed under a laser scanning confocal microscope after labeling with Mitotracker.The treated HUVECs were also labeled with JC-1 probe,and the mitochondrial membrane potential (MMP) was then examined by flow cytometry.The expression of mitophagy-related proteins including microtubule-associated protein 1 light chain 3 Ⅰ /Ⅱ (LC3 Ⅰ /Ⅱ),moesin-like BCL2-induced protein1(Beclin1),phosphatase and tensin (PTEN) homologinduced putative kinase 1(PINK1) and dynamin-related protein Ⅰ (DRP1) was determined by Westem blotting.RESULTS More than 95% of the cultured cells expressed vascular endothelial cadherin and herein were vascular endothelial cells.The MTT result showed that the cell survival of HUVECs was significantly decreased after incubation with CdTe QD (0.1-100 mg,L-1) for 24 h (P<0.05,P<0.01).CdTe QD also induced extensive fragmentation of the mitochondrial network.The results of JC-1 assay showed that CdTe QD (0.1-100 mg· L-1) caused the disruption of MMP.The percentage of HUVECs with higher MMP was reduced from (91.8±0.6)% in cell control group to (90.2±1.1)%,(84.4±0.9)% (P<0.05) and (78.1 ±1.3)% (P<0.01),respectively.The Western blotting data suggested that CdTe QD 10 mg·L-1 significantly increased the expression of autophagy-related protein beclin 1 and the ratio of LC3 Ⅱ/LC3 Ⅰ (P<0.05,P<0.01),CdTe QD 1 mg· L-1 also raised the level of mitophagy-related proteins like PINK1 and DRP1.CONCULSION CdTe QD can induce mitochondrial dysfunction as well as mitophagy in HUVECs.
5.DIFFERENT FACTORS AFFECTING ANTIBODY RESPONSES IN MICE IMMUNIZED BY GENE RECOMBINANT OF HCV STRUCTURE REGION
Jun DOU ; Kezhou LIU ; Zhi CHEN ; Jianer WO ; Nanxiang HE ; Yong LIU ; Mingtai ZHANG ; Xinzi WANG ; Chenghuai XU
Immunological Journal 1999;(3):151-155
To seek the optimum experiment methods of animal immunization with HCV gene and to explore the effect on antibody responses in mice immunized by pCD-HCV1 recombinant in different administration, recombinant pCD-HCV1 was constructed by technique of molecular biology and was injected into muscles of Balb/c of mice with different times, routes and dosage of inoculations as well as different treatment. The results showed that the serum antibody level reached 0.183±0.06,0.428±0.05,0.707±0.08 and 0.773±0.07(OD410 value) respectively after recombinant pCD-HCV1(100μg/mouse) were injected into mice once, twice, three times and four times. The antibody level of mice (n=12) with four times inoculation was the highest; pCD-HCV1 was perfused into stomach orally in mice or were into mice by i.p, s.c and i.m(100μg/mouse, three times) in different routes (n=6), and the antibody levels were 0.138±0.05, 0.178±0.07, 0.233±0.08 and 0.691±0.05 respectively; after the mice (n=8) were inoculated with the pCD-HCV1 of different dosage(10μg, 50μg and 100μg) the antibody levels of three groups were 0.11±0.09, 0.33±0.04, and 0.700±0.07, and the results showed a significant difference (P<0.01); Mice was injected with procaine (100μl, 0.4mg) by i.m or s.c. Then pCD-HCV1 was injected into mice and antibody levels were higher than that of mice immunized directly with recombinant pCD-HCV1 of same dosage. The results may provide a reference data deserved for screening the optimum immunization method of development HCV-DNA-based vaccine in mice model.
6.Research and application advances in rehabilitation assessment of stroke.
Kezhou LIU ; Mengjie YIN ; Zhengting CAI
Journal of Zhejiang University. Science. B 2022;23(8):625-641
Stroke has a high incidence and disability rate, and rehabilitation is an effective means to reduce the disability rate of patients. To systematize rehabilitation assessment, which is the foundation for rehabilitation therapy, we summarize the assessment methods commonly used in research and clinical applications, including the various types of stroke rehabilitation scales and their applicability, and related biomedical detection technologies, including surface electromyography (sEMG), motion analysis systems, transcranial magnetic stimulation (TMS), magnetic resonance imaging (MRI), and combinations of different techniques. We also introduce some assessment techniques that are still in the experimental phase, such as the prospective application of artificial intelligence (AI) with optical correlation tomography (OCT) in stroke rehabilitation. This review provides a useful bibliography for the assessment of not only the severity of stroke injury, but also the therapeutic effects of stroke rehabilitation, and establishes a solid base for the future development of stroke rehabilitation skills.
Artificial Intelligence
;
Humans
;
Magnetic Resonance Imaging
;
Stroke
;
Stroke Rehabilitation/methods*
;
Transcranial Magnetic Stimulation/methods*
7.Clinical efficacy of radical resection with individualized surgical approach for borderline resectable pancreatic head carcinoma
Chunlu TAN ; Hongyu CHEN ; Kezhou LI ; Hao ZHANG ; Xubao LIU
Chinese Journal of Digestive Surgery 2019;18(7):662-667
Objective To explore the clinical efficacy of radical resection with individualized surgical approach for borderline resectable pancreatic head carcinoma.Methods The retrospective descriptive study was conducted.The clinicopathological data of 54 patients with borderline resectable pancreatic head carcinoma who underwent radical resection with individualized surgical approach in the West China Hospital of Sichuan University from January 2015 to January 2018 were collected.There were 37 males and 17 females,aged from 37 to 73 years,with a median age of 59 years.For venous type borderline resectable pancreatic head carcinoma,surgery for pancreatic head carcinoma and (or) pancreatic head and neck carcinoma was performed via inferior mesenteric vein,and surgery for pancreatic uncinate process carcinoma was performed via inferior colon artery.For arterial type borderline resectable pancreatic head carcinoma,surgery for pancreatic head carcinoma and (or) pancreatic head and neck carcinoma was performed via medial uncinate artery,and surgery for pancreatic uncinate process carcinoma was performed via left posterior artery.Observation indicators:(1) surgical situations;(2) postoperative complications;(3) postoperative pathological examination;(4) follow-up.Patients were followed up by outpatient examination or telephone interview once every 3 months to detect survival up to March 2019.Measurement data with normal distribution were represented by Mean ± SD.Measurement data with skewed distribution were represented by M (range),and count data were represented by absolute numbers or percentage.Kaplan-meier method was used to draw the survival curve and calculate the survival rate.Results (1) Surgical situations:all the 54 patients underwent expanded pancreatoduodenectomy combined with superior mesenteric vein/portal vein (SMV/PV) resection,including 15 via inferior mesenteric vein,20 via inferior colon artery,12 via medial uncinate artery,and 7 via left posterior artery.The operation time was (320± 83)minutes,and the volume of intraoperative blood loss was (865±512) mL.(2) Postoperative complications:of 54 cases,28 had postoperative complications,including 13 with grade 1 Clavien-Dindo complications,12 with grade 2 ClavienDindo complications,3 with grade 3 or above Clavien-Dindo complications.One of the 28 patients with postoperative complications died and 27 were improved after symptomatic and supportive treatment.(3) Postoperative pathological examination:of 54 patients,31 had R0 resection and 23 had R1 resection.In the 23 patients with R1 resection,5 underwent surgery via the inferior mesenteric vein (4 with involvement of pancreatic anterior surface,1 with involvement of both pancreatic anterior and posterior surface),9 underwent surgery via the inferior colon artery (2 with involvement of both pancreatic anterior and posterior surface,2 with involvement of superior mesenteric artery margin,2 with involvement of pancreatic posterior surface,2 with involvement of pancreatic anterior surface,1 with involvement of superior mesenteric artery margin and pancreatic posterior surface),5 underwent surgery via the medial uncinate process artery (2 with involvement of superior mesenteric artery margin,2 with involvement of both pancreatic anterior and posterior surface,1 with involvement of pancreatic neck transected margin),and 4 underwent surgery via the left posterior artery (3 with involvement of superior mesenteric artery margin,1 with involvement of both pancreatic anterior and posterior surface).Of 54 patients,16 had no positive lymph nodes,26 had 1-3 positive lymph nodes,and 12 had 4 or more positive lymph nodes.The tumor diameter was (3.20±0.14)cm.There were 48 of 54 patients with nerve infiltration,41 with superior mesenteric vein and/or portal vein infiltration,and 11 with vascular thrombus.There were 17 of 54 patients with high differentiation and medium differentiation,and 37 with low differentiation and undifferentiation.(4) Follow-up:54 patients were followed up for 1-42 months,with a median time of 19 months.The 1-,3-year overall survival rate was 78.0%,11.4%.Condusion As for the borderline resectable pancreatic head cancer,individualized and customized surgical approach according to the location of tumor and the relationship with blood vessels is helpful to standardize the radical resection and avoid R2 resection.
8.Research progress on antiviral therapy in severe hepatic impairment patients and special population with HCV infection
Chinese Journal of Clinical Infectious Diseases 2018;11(5):339-345
The research and development of direct-acting antiviral agents (DAAs) for hepatitis C achieved significant results in the last decade .DAAs is highly effective in treatment of hepatitic C patients with convenience and safety , and the sustained virological response of DAAs is over 90%, which will greatly contribute to achieving the WHO's goal of eliminating hepatitis C by 2030.The European Association for the Study of the Liver (EASL), American Association for the Study of Liver Diseases (AASLD) and WHO all updated guidelines for diagnosis and treatment of hepatitis C in 2018, which will play a positive role for the management of hepatitis C globally .This article reviews the related guidelines and literature on prevention and treatment of hepatitis C , and summarizes the advantages of antiviral therapy for severe hepatic impairment patients and special population with HCV infection .
9.Expression of IL-2/IL-15Rβ on memory CD3 + CD8 + CD45RO + T cells in chronic hepatitis B patients undergoing antiviral treatment and its significance
Wenzheng HAN ; Qing CHEN ; Jianghua YANG ; Xiaoxin WU ; Kezhou LIU ; Xiaoning LI
Chinese Journal of Clinical Infectious Diseases 2021;14(2):107-115
Objective:To investigate the expression of IL-2/IL-15 receptor β subunit (IL-2/IL-15Rβ) on memory CD3 + CD8 + CD45RO + T cells in patients with chronic hepatitis B (CHB) receiving antiviral treatment and its significance. Methods:Sixty-eight patients with chronic active hepatitis B (CAHB) and 47 asymptomatic hepatitis B virus (HBV) carriers attending in the Department of Infectious Diseases, the First Affiliated Hospital of Wannan Medical College from March 2019 to December 2020 were enrolled in the study; and 30 health subjects were also enrolled as healthy control group. Among 60 CAHB patients there were 30 cases with positive HBeAg and 30 cases with negative HBeAg. All CAHB patients received nucleos(t)ide analogue therapy, the HBV-related markers, Alanine aminotransferase (ALT) and the expression of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were determined and compared between HBeAg-positive and negative patients, before and after treatment. Normal distribution measurement data among 3 groups were compared with One-way ANOVA; normal distribution measurement data between 2 groups were compared with paired samples t test; non-normal distribution measurement data between the two groups were compared with Mann-Whitney U test; Pearson’s correlation coefficient was performed for correlation analysis. P<0.05 was considered statistically significant. Results:The proportion of CD8 + CD45RO + T cells on PBMC CD3 + T cells in CAHB group [(8.6±3.7)%] was higher than that of asymptomatic HBV carriers group [(5.7±2.5)%] and healthy control group [(5.5±1.5)%] (all P<0.05). The expression percentage of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells in CAHB group [(6.8±4.7)%] was higher than that of asymptomatic HBV carriers group [(4.7±2.8)%] and healthy control group [(4.3±2.2)%] (all P<0.05). The MFI of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells in CAHB group (243±168) was higher than those of asymptomatic HBV carriers group (160±91) and healthy control group [160±63] (all P<0.05). The expression percentage and MFI of IL-2/IL-15Rβ on PBMC CD3 + CD8 + CD45RO + T cells were positively correlated with the percentage of CD3 + CD8 + CD45RO + T cells in CAHB patients ( r=0.33 and 0.28, all P<0.05). The proliferation percentage of PBMC CD3 + CD8 + CD45RO + T cells in CAHB group[ (43.7±16.0)%] was higher than that of asymptomatic HBV carriers group [(29.1±9.4)%] and healthy control group [(26.8±9.6)%] after stimulation with Anti-CD3+ super-2 (all P<0.05). After the expression of IL-2/IL-15Rβ was blocked, the proliferation percentage of CD3 + CD8 + CD45RO + T cells was decreased [(11.2±6.3)%] compared with the untreated CAHB group ( P<0.05). The percentages of PBMC CD3 + CD8 + CD45RO + T cells secreting IFN-γ, IL-2 and TNF-α in CAHB group were (13.8±5.4)%, (14.0±4.3)% and (12.3±4.6)% respectively, which were higher than those of asymptomatic HBV carriers [(8.4±2.6)%, (9.4±3.2)% and (6.8±3.3)%] and healthy control group [(6.9±2.7)%, (9.9±3.0)% and (7.7±3.8)%] after stimulation with Anti-CD3+ super-2 (all P<0.05). After the expression of IL-2/IL-15Rβ was blocked, the percentages of PBMC CD3 + CD8 + CD45RO + T cells secreting IFN-γ [(2.4±1.6)%], IL-2 [(4.1±1.9)%] and TNF-α [(4.1±1.8)%] were decreased compared with the untreated CAHB group (all P<0.05). HBeAg, ALT, the expression percentage and MFI of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were 521.4 (68.9, 1 339.0) COI, 292 (160, 528) U/L, (6.4±3.2)% and (239±136) in 30 HBeAg-positive CAHB patients before treatment, which were higher than those after treatment [3.5(1.5, 17.5)COI、20(14, 31) U/L, (4.1±2.4)% and (134±58)] ( Z=5.337 and 6.403, t=3.229 and 3.892, all P<0.05). HBsAg, ALT, the expression percentage and MFI of IL-2/IL-15Rβ on CD3 + CD8 + CD45RO + T cells were (5 310±2 851) COI, (328±207) U/L, (7.1±5.8)% and (252±110) in 30 HBeAg-negative CAHB patients before treatment, which were higher than those after 48 weeks of treatment [(3 811±2 495) COI, (33±14) U/L, (4.6±2.9)% and (154±73)] ( t=2.167, 5.595, 2.116 and 2.383, all P<0.05). Conclusion:The study suggests that up-regulated expression of IL-2/IL-15Rβ is associated with elevated frequency, proliferation and secretion function of memory CD3 + CD8 + CD45RO + T cells in CAHB patients.