1.Mass spectrometry technology and its application in analysis of biological samples.
Longshan ZHAO ; Qing LI ; Chaowei GUO ; Xiaohui CHEN ; Kaishun BI
Acta Pharmaceutica Sinica 2012;47(2):158-62
With the excellent merits of wide analytical range, high sensitivity, small sample size, fast analysis speed, good repeatability, simple operation, low mobile phase consumption, as well as its capability of simultaneous isolation and identification, etc, mass spectrometry techniques have become widely used in the area of environmental science, energy chemical industry, biological medicine, and so on. This article reviews the application of mass spectrometry technology in biological sample analysis in the latest three years with the focus on the new applications in pharmacokinetics and bioequivalence, toxicokinetics, pharmacokinetic-pharmacodynamic, population pharmacokinetics, identification and fragmentation pathways of drugs and their metabolites and metabonomics to provide references for further study of biological sample analysis.
2.Detection of immune cell subsets in renal allograft recipients before operation and its significance
Dongwei LI ; Longshan LIU ; Jiguang FEI ; Changxi WANG
Chinese Journal of Tissue Engineering Research 2013;(44):7675-7680
BACKGROUND:The immune cells of renal al ograft recipients have always been the hot spot of research. However, there are few studies addressing the immune cellsubsets in renal al ograft recipients before operation. OBJECTIVE:To investigate the proportional distribution of immune cellsubsets in renal al ograft recipients before operation. METHODS:Fifteen de novo living-related renal transplant recipients were enrol ed in this study with 15 healthy volunteers, aged 18-40 years, as healthy controls. Flow cytometry was employed to observe the proportion of the immune cellsubsets by extracting peripheral venous blood of al participants. RESULTS AND CONCLUSION:In the renal al ograft recipients, the proportions of CD4+CD25+T cells, the proportion of CD4+CD25+/CD4+T cells, CD19+B cells, CD19+CD5+B cells, CD19+CD27+B cells, NKG2A/NK cells, and NKG2A/NKG2 cells were al lower than those in the healthy controls;however, the proportion of CD38+IgD-/CD19+B cells and NKG2D cells were higher than those in the healthy controls. The difference of the proportion of immune cellsubsets aforementioned between the two groups was statistical y significant (P<0.05), while no difference was observed in other subsets. Immune cellsubsets in renal al ograft recipients before operation could be used to assess the immune status of the recipients, and also could be seen as the basal control for postoperative immunological monitoring.
3.Effect of Converted Regulatory T Cells with Renca Conditioned Media on Suppressive Profile of Allo-immunity in Mice
Lichen TENG ; Longshan LIU ; Yajuan SU ; Xiaopeng YUAN ; Jun LI ; Qian FU ; Siyang CHEN ; Changxi WANG
Journal of Sun Yat-sen University(Medical Sciences) 2010;31(2):242-248
[Objective]This study was designed to investigate capability of the conditioned media that originated from Renca cells to convert CIM~+ CD25~- T cells into CD4~+ CD25~+ T cells,which can exert immunosuppressive effect on effector T cells in vitro and in vivo.[Methods]The common media were mixed with the conditioned media at different ratios,and fresh enriched CD4~+ CD25~- T cells with MACS were cultured in mixed media for 7 days.At end-point of culture,the cells were collected and detected phenotypes in flow cytometer.Moreover,we detected immunosuppressive effect of converted CD4~+ CD25~+ T cells on effeetor T cells proliferation in one-way mixed lymphocytes reaction by using CCK-8,and we observed survival time and histology of grafts.The delayed type hypersensitivity was determined 14 days after transplantation.[Results]The mixed media could increase ratio of CD4~+ CD25~+ Foxp3~+ T cells in conditioned media ratio-dependent(P<0.05),compared with control groups,when the mixed media contained no mote than 75% of conditioned media.The converted CD4~+ CD25~+ T cells significantly suppress proliferation of effector T cells in vitro,and prolong survival time of grafts,which were(29.6±1.4)d in converted CD4~+ CD25~+ T cells treated groups(P<0.05),compared with that in untreated groups(9.8±0.6 d)or PBS treated groups(10.9±0.6 d).Moreover,delayed-type hypersensitivity reaction were conducted at day 14 after transplantation in the recipients,and the results showed that less pad swelling in the group treated with converted CD4~+ CD25~+ T cells than other control groups was found,according to measurement of pad swelling.In addition,progressed to complete necrosis of grafts were exhibited in the mice treated with PBS and untreated mice,whereas better healing of grafts and less lymphocytes infiltration were displayed in the mice treated with converted CD4~+ CD25~+ T cells,which were similar to the mice treated with natural regulatory T cells.[Conclusion]The converted CD4~+ CD25~+ T cells with Renca conditioned media play suppressive role in vitro and in vivo.
4.Morphological Observation of Haplorchis pumilio
Jinxiang LIN ; Baojian CHEN ; Kai ZHU ; Youzhu CHENG ; Yousong LI ; Longshan XU ; Rongyan ZHANG
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(06):-
Objective To provide the basis for the differentiation with similar species of intestinal flukes through observing the figure of Haplorchis pumilio. Methods Adults of H. pumilio were collected from the intestine of the cat which was infected with the encysted cercariae of H.pumilio for 45 days. The worms were observed after staining. Eggs and metacercariae of H.pumilio were collected and examined for their shape, size and morphological characteristics. Pseudorasbora parva, the fish host, was examined for the parasitized sites of metacercariae. Results The principal characteristics of the adults is the acetabulum degradation. There are only the genital sucker with 44-48 hamuli. The average measurement of eggs is 31.2?16.7 ?m with a smooth shell. Its aceromion is not evident. The average diameter of metacercariae is 168.5 ?m. There are squamous spines on metacercaria. The metacercariae only parasitize in the muscle between the basis of the fin and the fish body. The average measurement of metacercaria cyst is 445?95?m, with squamous spines on the body surface. Hamuli are found on the genital sucker of metacercaria cyst. Conclusion The morphological figures and parasitic sites of metacercaria, the genital sucker of the adult, and the number and form of the hamulus on the genital sucker provide basis for distinguishing H. pumilio from other intestinal flukes.
5.Genotype Detection of the Merozoite Surface Protein Alleles of Plasmodium vivax
Shanying ZHANG ; Longshan XU ; Huimin LU ; Yingzhen ZHANG ; Qi GAO ; Lisha LI
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(02):-
Objective To develop a method for detecting the genotype of Plasmodium vivax merozoite surface protein 1 (PvMSP-1) alleles. Methods According to the sequence characteristic of PvMSP-1, nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to amplify the polymorphic region of ICB5-ICB6 which contains Q repeats and PvuII restriction site (Sal-1 type). The PCR product was digested by PvuII restriction endonuclease and the digested fragments were observed by 2% agarose gel electrophoresis. The allelic type was determined according to the banding pattern. Results Bands in size of 400 bp (Belem type ) and/or 470 bp (Sal-1 type ) appeared in all 98 P. vivax isolates, no band was found in negative control. After PvuII digestion, two Sal-1 type fragments (120 bp and 350 bp) were obtained from 45 samples of 470 bp. Single-band of 400 bp appeared in 3 of 40 samples with 400 bp as Belem type, two bands of 120 bp and 280 bp appeared from other 35 samples as recombination type III, and another 2 bands with 120 bp and 240 bp as Korean isolate. Conclusion The result showed that the nested PCR-RFLP may be applied in the detection and identification of the three PvMSP-1 allelic types in China.
6.Single center experiences: characteristics of tuberculosis in iost-renal-transplant recipients during 20 years
Ling ZHANG ; Changxi WANG ; Hongmei FU ; Qian FU ; Jun LI ; Lizhong CHEN ; Jiguang FEI ; Suxiong DENG ; Longshan LIU
Chinese Journal of Organ Transplantation 2011;32(10):600-603
Objective To analyze the characteristics of tuberculosis (TB) in renal-transplant recipients from our hospital, and summarize the corresponding experiences in diagnosis and management.Methods A retrospective study was performed on 61 documented post-transplant TB cases out of the 2842 patients who received kidney transplantation in the First Affiliated Hospital of Sun Yat-sen University between Jan.1991 and Dec.2010.Results TB in the post-renal-transplant population in our hospital displayed the following characteristics:(1) High incidence (2.1% ).54.1% recipients were diagnosed within the first year post-transplant; (2) Lung was the most common site (77.0 %).There was high prevalence (60.7 %) of extra-pulmonary TB (lymphatic TB,23.0 %; pleuritis,13.1 %; graft,11.5%); (3) Fever (83.6 %),cough (55.7 %),sputum (41.0 %) were the most common clinical manifestations.There were also emaciation (3.3 %) and enlargement of lymph nodes (18.0 %); (4) Chest X-ray and CT were of great value during TB diagnosis while purified protein derivative of tuberculin (PPD) skin test had little diagnostic value with a negative result in 56 cases (91.8 %) ; (5) Liver function damage ( 16.4 %),kidney function injury (39.3 %) and peripheral nerve toxicity (3.3 %) were the main adverse reactions of anti-tuberculosis chemotherapy,also the major cause of anti-TB failure; (6) Pre-transplant TB (17 cases) increased the probability of TB recurrence (4 cases,23.5 %) post-transplantation; (7) The post-transplant TB patients were accompanied with cellular immune deficiency,resulting in overlapping infection of bacteria,viruses and fungi (19.7 %); (8) 1- and 3-year patient/graft survival rate of patients with post-transplant TB was 85.2 %/78.7 % and 85.2 %/75.4 % respectively. The accumulative mortality rate reached to 14.8%,while overlapping infection was the major cause of death (66.7 %).Conclusion Chinese renal transplant recipients still face a high risk of TB because of their immunecompromised state and epidemiological prevalence of the disease. For the high mortality rate and associated serious complications,rapid diagnosis and effective anti-TB chemotherapy are of great value for TB population.
8.Pharmacokinetics of doxapram injection in Chinese Han, Mongolian, Korean, Uigur and Hui healthy subjects
Tao GUO ; Longshan ZHAO ; Jia CHENG ; Dongya XIA ; Ping ZENG ; Yong XIAO ; Dongxiang ZHAO ; Qiang LI ; Yin BAI ; Changqing YANG
Chinese Pharmacological Bulletin 2014;(11):1595-1599
Aim To investigate and compare the phar-macokinetics of doxapram injection in healthy subjects of different Chinese nationalities including Han, Mon-golian, Korean, Hui and Uigur, and the influence of gender,in order to provide instruction and help for the usage of doxapram for both clinic and remedy of battle wound. Methods An HPLC-UV method was used to determine the plasma concentration of doxapram. Fifty healthy subjects ( five males and five females of each nationality) were recruited for the study. A single dose of 50 mg doxapram was administered intravenously to the healthy subjects, and blood samples were collected at various predetermined time points. The pharmacoki-netic parameters were calculated by DAS software and were compared by SPSS 13. 0 software, in order to as-sess the influence of nationality or gender on pharmaco-kinetics of doxapram. Results The results indicated that the pharmacokinetic profile of doxapram in vivo could be described as two-compartment model. The main pharmacokinetic parameters for Han, Mongolian, Korean, Hui and Uygur were as follows: Cl ( 0. 25 ± 0. 11 ) , ( 0. 33 ± 0. 11 ) , ( 0. 27 ± 0. 07 ) , ( 0. 26 ± 0. 06) and (0. 39 ± 0. 25) L·h-1 ·kg-1 , while Cmax (1. 55 ± 0. 52 ) , ( 1. 02 ± 0. 30 ) , ( 1. 31 ± 0. 47 ) , (1. 48 ± 0. 46 ) and ( 0. 99 ± 0. 35 ) mg · L-1 . The AUC0-12. 5 , AUC0-∞ and Cmax of Chinese Han were sig-nificantly higher than those of Uigur and Mongolian ( P<0. 05 ) , while there was no significant difference in other parameters ( P>0. 05 ) . There were statistically significant differences in Vc , Vd and CL between young males and females ( P < 0. 05 ) . Conclusion The large inter-individual variation in the main pharmacoki-netics suggests the dosage of doxapram should be ad-justed for different nationalities for both clinic and rem-edy of battle wound.
9.Formation mechanism of neutrophil extracellular traps and their role in liver diseases
Lin CAO ; Qian LI ; Longshan JI
Journal of Clinical Hepatology 2020;36(1):205-207
Neutrophil extracellular traps (NETs) have recently been found to be a new immune mechanism for neutrophils to kill pathogens. The main component of NETs is DNA, with granular proteins of different sizes attached to the surface. NETs play different roles in various liver diseases such as hepatic ischemia-reperfusion injury, chronic hepatitis B, liver cirrhosis, and liver cancer. This article reviews the formation mechanism, structure, and role of NETs in liver diseases.
10.Clinical efficacy of dual-kidney transplantation from infant donors to adult recipients
Mingchuan HUANG ; Chenglin WU ; Jun LI ; Xiaopeng YUAN ; Ronghai DENG ; Yitao ZHENG ; Longshan LIU ; Shenghui WU ; Xixi GAN ; Changxi WANG
Chinese Journal of Organ Transplantation 2021;42(1):8-13
Objective:To explore the clinical efficacy of dual-kidney transplantation from infant donors to adult recipients.Methods:From December 2012 to November 2020 in Organ Transplant Center First Affiliated Hospital Sun Yat-sen University, rertrospective reviews were conducted for clinical data of 25 pairs of infant donors and adult recipients. The survival rates were calculated for both recipients and transplanted kidneys at Year 1/3/5 post-transplantation. And the postoperative recovery status and the postoperative incidence of adverse events of recipients were observed.Results:The survival rates of recipients were all 95.8% at Year 1/3/5 and those of transplanted kidney and dealth-cancelling transplanted kidney all 87.2%. One case died due to acute inferior-wall cardiac infarction while three others lost renal functions for vascular thrombosis, ureteral stenosis and urinary fistula. Except for loss of renal function and death, the postoperative estimated golmerular fitration rate was (99.35±21.78), (103.11±29.20) and (114.99±28.55) ml/(min·1.73 m 2) at Year 1/2/3 respectively. Conclusions:Selecting proper recipients, standardizing donor acquisition and surgical procedures and strengthening perioperative managements may expand the donor pool. The overall outcomes are excellent for adult recipients with dual-kidney transplantation from donations after infants' death.