1.Roles of pericytes in the urological and reproductive systems.
Ming-Kuan ZHOU ; Ya-Dong ZHANG ; Xiang-An TU
National Journal of Andrology 2014;20(12):1126-1130
Pericyte, also known as mural cell or Rouget cell, is one of the main cells that make up the wall of capillaries. Pericytes play important roles not only in the maturation, stability, and function maintenance of blood vessels, but also in the growth and development of tissues and organs, wound repair, and other physiological and pathological processes. Researches on the functions of pericytes are mainly concentrated on their multipotency, adjustment of vascular functions, and process of fibrosis, as well as on renal fibrosis, renal blood flow regulation, and glomerular filtration in urology, but are quite insufficient in andrology. This article reviews the location, origin, distribution, morphology, markers, and functions of pericytes, aiming to induce further studies of pericytes in andrology.
Fibrosis
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pathology
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Humans
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Pericytes
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physiology
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Urogenital System
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cytology
2.Protective effect of Rheum tanguticum polysaccharides (RTP) on traumatic brain injury in rats.
Zhi-peng WANG ; Li LIU ; Qi-bing MEI ; Rong ZHANG ; Jian-wen GU ; Xiang ZHANG ; Da-kuan GAO
China Journal of Chinese Materia Medica 2003;28(10):974-971
OBJECTIVETo evaluate protective effects of Rheum tanguticum polysaccharides (RTP) on traumatic brain injury (TBI) in rats.
METHODThe polysaccharides (RTP) were extracted from Tanguficum Maxim. 120 rats were divided into 15 groups, with 8 rats in each group. RTP at 100, 200 and 400 mg x kg(-1) were administrated orally once a day for five days, and model of brain injury was made by dropping weight method.
RESULTRTP reduced water content and malondialdehyde (MDA) levels, and increased total SOD activity and Na+-K+ ATPase activity after injuried.
CONCLUSIONThe polysaccharides may be one of the effective comptents in Rheum tanguticum, showing significant neuroprotective effects.
Animals ; Brain Injuries ; enzymology ; metabolism ; pathology ; Cerebral Cortex ; enzymology ; ultrastructure ; Male ; Malondialdehyde ; metabolism ; Neuroprotective Agents ; pharmacology ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism
3.Human bone marrow mesenchymal stem cells have little preventive or therapeutic effect on rat arthritis induced by collagen.
Yong-Qi WANG ; Xiao-Fang CAO ; Zi-Ming HE ; Feng-Jun XIAO ; Heng-Xiang WANG ; Zi-Kuan GUO
Journal of Experimental Hematology 2012;20(2):433-437
The aim of this study was to investigate if transfusion of mesenchymal stem cells (MSC) could exhibit beneficial effects on rheumatoid arthritis. Human bone marrow MSC were intraperitoneally injected into Wistar rats with collagen-induced arthritis at a dose of 10(7) on the next day (preventive group) or 2 weeks (treatment group) after collagen II induction, once a week for 2 weeks (preventive group) or 4 weeks (treatment group). The control group was given normal saline (NS) at corresponding time. The symptom scorings were documented weekly from the second week of the induction. On week 6, the hind joints of the rats were pathologically examined and the activation status of splenocytes was analyzed by flow cytometry. The results showed that all the rats developed arthritis and subsequent joint abnormality. On the sixth week, symptom scores of the rats that received MSC preventive (9.5 ± 0.5) or therapeutic (9.4 ± 0.6) infusions had no significant difference between each other, but were significantly greater than those of the NS controls (7.6 ± 0.6, P < 0.05). Consistently, pathological examination on the involved knees showed that the synovitis and arthritis scorings of MSC treated rats were greatly elevated compared with NS controls. Furthermore, the ratios of CD86(+) cells in the spleens of MSC prevention, MSC treatment and NS control groups were (4.16 ± 1.48), (4.06 ± 1.97) and (4.15 ± 2.04) respectively, while those of CD11b/c(+)CD86(+) cells were (1.04 ± 0.68), (0.95 ± 0.56) and (0.98 ± 0.44), all of which were significantly higher than those of healthy controls [(0.97 ± 0.18) and (0.30 ± 0.17), P < 0.05 for both parameters]. It is concluded that MSC infusion has little beneficial effects on collagen-induced arthritis in rats, conversely, MSC therapy aggravated the damage of the involved joints, its underlying mechanisms need to be further investigated.
Animals
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Arthritis, Experimental
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pathology
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prevention & control
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therapy
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Bone Marrow Cells
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Cells, Cultured
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Humans
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Male
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Mesenchymal Stem Cell Transplantation
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Mesenchymal Stromal Cells
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cytology
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Rats
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Rats, Wistar
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Transplantation, Heterologous
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Treatment Outcome
4.Production of recombinant humanized anti-HBsAg Fab antibody by fermentation.
Ning DENG ; Jun-Jian XIANG ; Wen-Yin CHEN ; Sheng XIONG ; Xun-Zhang WANG ; Kuan-Yuan SU
Chinese Journal of Biotechnology 2004;20(5):800-804
In order to produce recombinant human anti-HBsAg Fab antibody in Pichia pastoris, the recombinant yeast was fermented using fed-batch system in a 30 L bioreactor. The fermentation temperature was 30 degrees C, the pH was 5.0 approximately 5.3, and the DO was 20% approximately 30%. The recombinant Fab antibody was purified from crude culture supernatant by ion exchange and analyzed by SDS-PAGE and western blot and ELISA. When the absorbance (OD600) of broth reach 300 at the end of fed-batch phase, the induced phase was initiated. The results showed that recombinant human anti-HBsAg Fab antibody was high-level expressed in recombinant Pichia pastoris using a fed-batch fermentation system. Both chains of the Fab were successfully expressed upon methanol induction. After 192 h of induction, the expression level of recombinant Fab (soluble) reached 412 mg/L. The recombinant Fab antibody was purified effectively by ion-exchange chromatography from the fermentation supernatant to a purity of 95%. And the affinity activities of the purified recombinant Fab antibdy and fermentation supernatant were detected, and both of them showed high affinity activities. The results demonstrated that recombinant human anti-HBsAg Fab antibody could be high level produced by fed-batch fermentations in Pichia pastoris. Which can be efficiently used in industrial production.
Fermentation
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Hepatitis B Antibodies
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biosynthesis
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isolation & purification
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Hepatitis B Surface Antigens
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immunology
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Humans
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Immunoglobulin Fab Fragments
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biosynthesis
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isolation & purification
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Pichia
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genetics
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Recombinant Proteins
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biosynthesis
;
isolation & purification
5.Effects of tumor necrosis factor-α on immunoregulatory activities of mesenchymal stem cells in vitro and in vivo.
Yong-Qi WANG ; Zhi-Yong LI ; Xiao-Fang CAO ; Heng-Xiang WANG ; Zi-Kuan GUO
Journal of Experimental Hematology 2012;20(4):981-984
Mesenchymal stem cells (MSC) are characterized by their potent immuno-regulatory activity, however our previous data have shown that MSC have no therapeutic effects on collagen-induced arthritis (CIA). To further clarify the complexity, the effects of tumor necrosis factor-alpha (TNF-α) on the in vitro and in vivo immunoregulatory activity of MSC were investigated in this study, as TNF-α is recognized as the key factor in the development of rheumatoid arthritis. The nuclear translocation of the inflammation-associated factor NF-κB was observed after human umbilical cord MSC were treated with TNF-α and the cell proliferation status was assessed by MTT test. The inhibitory effects of MSC or TNF-α-treated MSC on the mixed lymphocyte reaction, in which Wistar rat spleen mononuclear cells were served as the responders and the splenocytes from SD rat spleens as the stimulators, were also determined by the MTT test. Further, the therapeutic potentials of MSC or TNF-α-treated MSC were observed in a Wistar rat CIA model. The results showed that NF-κB translocated into the nuclei promptly after TNF-α treatment, though TNF-α had little effect on the MSC proliferation. MSC, whether pre-stimulated by TNF-α or not and when different doses were tested, exhibited obviously inhibitory effects on the proliferation of the lymphocytes (P < 0.001 for all groups tested), while MSC-treated by TNF-α displayed more potent suppression especially when low-density were used. Unexpectedly, the infiltration of inflammatory cells into the involved knees was aggravated by cell treatment and the pathological scores were significantly higher than those of controls (P < 0.05). It is concluded that the TNF-α exhibits different effects on immune regulation activity of MSC, and its underlying mechanism needs to further investigate.
Animals
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Arthritis, Experimental
;
metabolism
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pathology
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Cell Proliferation
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drug effects
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Cells, Cultured
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Humans
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Male
;
Mesenchymal Stromal Cells
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cytology
;
drug effects
;
immunology
;
NF-kappa B
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metabolism
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Rats
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Rats, Sprague-Dawley
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Rats, Wistar
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Tumor Necrosis Factor-alpha
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pharmacology
6.Therapeutic effect of human bone marrow mesenchymal stem cell lysates on rat arthritis induced by collagen.
Xiao-Fang CAO ; Yong-Qi WANG ; Zhi-Yong LI ; Zi-Kuan GUO ; Heng-Xiang WNAG
Journal of Experimental Hematology 2013;21(3):765-769
Our previous work has shown that mesenchymal stem cells (MSC) have little therapeutic effect on rat arthritis induced by collagen. This study was aimed to further investigate whether the MSC lysates exhibit beneficial effects on rheumatoid arthritis. Aliquots of cell lysates from 1×10(7) human bone marrow MSC were intraperitoneally injected into collagen-induced arthritis (CIA) Wistar rats weekly for 4 consecutive weeks. Methotrexate at a dose of 1 mg/kg or normal saline was served as positive and negative controls respectively. On week 4 the symptom scores were recorded and the hind joints of the rats were pathologically examined and X-ray examination was performed. The results showed that on week 4, the symptom scores of the rats that received MSC lysates (6.87 ± 0.83) and MTX (6.44 ± 1.13) were significantly lower than that of control rats (7.33 ± 0.77, P < 0.01). Meanwhile, pathological examination on the involved ankle showed that the synovitis and arthritis scores of MSC lysates and control groups were 2.28 ± 0.48 and 2.28 ± 0.55 respectively, significantly higher than that of MTX treatment rats (0.71 ± 0.48, P < 0.05). However, X-ray examination on the ankle joints showed that the injury score of control rats was 4 ± 0.57, greatly higher than those from MSC lysates (2.71 ± 0.75) and MTX treatment groups (2.57 ± 0.78, P < 0.05 for both groups). It is concluded that MSC lysate infusion has beneficial effects on CIA rat, but the effectiveness seems inferior to MTX.
Animals
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Arthritis, Experimental
;
chemically induced
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therapy
;
Bone Marrow Cells
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cytology
;
Cells, Cultured
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Collagen
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Humans
;
Male
;
Mesenchymal Stromal Cells
;
cytology
;
Methotrexate
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pharmacology
;
Rats
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Rats, Wistar
7.Interspecific association of companion species in Artemisia annua community in southwest of China.
Ye-Kuan WU ; Xiang LIU ; Long-Yun LI ; Rui PENG ; Guang-Lin CUI ; Ling YUAN
China Journal of Chinese Materia Medica 2013;38(5):670-673
The interspecific association of companion species in Artemisia annua community in 48 region of southwest China was analyzed by variance analysis, chi2-test and association indices. The total related variance ratio among species in A. annua community was 2.05. Among 45 species pairs of 10 main species in the community, only 4 pairs showed significant negative correlations. Chi2 test, PC, OI, DI and AC values indicated pairs 1-8 (A. annua- A. lactiflora), 1-9 (A. annua- Setaria viridis) and 1-10 (A. annua- Bidens pilosa) showed a high correlations, and common utilization to non-restrictive resources. The results indicated that there was a significant positive correlation among species,and the community was at a stable stage, showed strong ability to human interference.
Analysis of Variance
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Artemisia annua
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China
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Conservation of Natural Resources
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Drugs, Chinese Herbal
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Symbiosis
8.Application of PCR-RFLP in detection of adefovir dipivoxil resistance-associated mutations in hepatitis B virus
Ri-Cheng MAO ; Ji-Ming ZHANG ; You-Kuan YIN ; Yan-Li QIN ; Wan-Qin ZHANG ; Xiang-Hui WU ; Xin-Hua WENG ;
Chinese Journal of Infection and Chemotherapy 2007;0(05):-
Objective To establish a convenient,accurate and practical method for detection of adefovir dipivoxil resistance-as- sociated mutation in hepatitis B virus:rtA181V/T/S and rtN236T mutations.Methods According to HBV complete sequences in GenBank,two pairs of primers were designed to amplify the region of HBV reverse transcriptase in order to introduce a BglI restriction site upon PCR product of wild type (wt) and a BseDI restriction site upon PCR product of rt236 mutant type.After amplification,the PCR products were digested with BglI and BseDI separately.We used this method to detect wild,rt181 mu- tant,rt236 mutant plasmids and 3 chronic hepatitis B patients' serum with obvious ADV resistance-associated mutations.We also tested the sensitivity of this method by mixing the wild and mutant plasmids in different proportions.Results The method could detect rt181 and rt236 mutations simultaneously.The result of RFLP analysis was in accordance with that of DNA se- quencing and cloning analysis.This method could detect the mutants even when they comprised only 10% of the total virus population.Conclusions The PCR-RFLP method with high sensitivity can detect rt181 and rt236 mutations simultaneously.It can be used for early detection of ADV resistance-associated mutation in hepatitis B virus.
9.Heme oxygenase-2 gene deletion protects basal ganglia cells from oxidative injury induced by free Fe2+
Yan QU ; Xiang ZHANG ; Jian-Ning ZHANG ; Hai-Ning ZHEN ; Da-Kuan GAO
Academic Journal of Second Military Medical University 2005;26(4):390-395
Objective:To determine whether heme oxygenase-2(HO-2) gene deletion can attenuate oxidative injury induced by free Fe2+. Methods:Stereotactic injection of 10 μl sterile FeCl2 (10 mmol/L) was made into the right striata of HO-2 knockout mice and wild-type mice. Brain edema severity was measured at 24 h. Cell viability, protein oxidation, and lipid oxidation of the basal ganglia were determined at 72 h. Western blot analysis was applied for heme oxygenase-1 (HO-1) measurement.Results: Brain water content significantly decreased in HO-2 knockout mice at 24 h compared with wild-type mice. Protein oxidation and lipid oxidation significantly decreased in HO-2 knockout mice at 72 h compared with wild-type mice, while the striatal cell viability increased significantly. HO-1 expression at baseline and 72 h was also similar to that in wild-type mice. Conclusion:These results show that HO-2 gene deletion can protect basal ganalia cells from free Fe2+ -mediated oxidative stress injury,suggesting that selective inhibition of HO-2 may have a protective effect on brain oxidative injury.
10.Expression of a human single-chain Fv antibody against HBsAg in Pichia pastoris.
Sheng XIONG ; Xiang-Rong REN ; Yong-Hong TANG ; Kuan-Yuan SU ; Zhou-Yao YU ; Yong LUO ; Yi-Fei WANG ; Jiu-Xiang LI
Chinese Journal of Biotechnology 2003;19(1):19-23
To express and secrete native HBscFv (anti-HBsAg single-chain Fv) in P. pastoris, HBscFv was amplified from plasmid pGEM-HBscFv, and then sub-cloned into expression vector pPICZalphaA. The resulting plasmid pPIC-HBscFv was linearized and transformed into P. pastoris GS115. The recombinant Pichia strains, identified by direct PCR and Zeocin-resistant screening of Pichia transformants, were cultured and induced with methanol. It was found that recombinant HBscFv, lead by alpha-factor, could be secreted into the culture supernatant to a level of 80mg/L. The bioactivity of Pichia produced HBscFv was confirmed by indirect ELISA, which also suggested that the bioactivity of HBscFv in the culture supernatant reached its peak in 72h and decreased in the late-stage of the induction. PAS staining suggests that HBscFv produced by yeast is poorly glycosylated or none-glycosylated protein.
Blotting, Western
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Electrophoresis, Polyacrylamide Gel
;
Hepatitis B Surface Antigens
;
immunology
;
Humans
;
Pichia
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genetics
;
metabolism
;
Polymerase Chain Reaction
;
Single-Chain Antibodies
;
genetics
;
immunology
;
metabolism