1.Study on Optimizing Preparative Procedure for Isolating and Purifying the Soluble Collagen Type Ⅱ
Xugan JIANG ; Shengxia CHEN ; Huifang WANG ; Liang WU ; Huaxi XU
Chinese Journal of Clinical Laboratory Science 2006;0(06):-
Objectives To establish a reliable method for isolating and purifying soluble collagen type Ⅱ(SCⅡ)by optimizing preparative procedure.Method The chicken sternal cartilage was selected as raw material.Guanidine hydrochloride was used to remove the proteoglycans.The digestion manners of pepsin,sodium chloride concentrations for salting,types of DEAE anion resin were studied for extracting SCⅡ.The SCⅡ identification was made by SDS-PAGE,absorption spectrum and amino acid analysis.Result It was convenient for pre-treatments of chicken sternal cartilage.The proteoglycans could be efficiently removed by 4 mol/L guanidine hydrochloride.The satisfied results were obtained by limited enzyme digestion of pepsin added by two steps.The optimizing concentration of sodium chloride for salting was 2.4mol/L.SDS-PAGE maps revealed that the bands of purified SCⅡand standard CⅡ were at the same location.The absorption peak of SCⅡ was at 230nm.The concentrations of Gly,Pro and Ala were the highest in 15 amino acids.Conclusion The improved method has significant advantages of simple working process,result reliability and convenient source of raw material.It is suitable for purifying the SCⅡ at variable scales in research works and clinic application.The SCⅡ product obtained has high purity and accords with the characteristics of collagen type Ⅱ.
2.Increased frequency of Th17 cell in patients with Hashimoto's thyroiditis
Yinzhao LIU ; Chenlu ZHU ; Jie TIAN ; Jianghua CHEN ; Jianguo CHEN ; Juan CHEN ; Huaxi XU ; Shengjun WANG
Chinese Journal of Endocrinology and Metabolism 2012;28(8):638-640
Objective To study the alteration of Th17 cells and related molecules in patients with Hashimoto's thyroiditis (HT).Methods Th17 cells were determined by flow cytometry.Real-time PCR method was applied to detect the expression of the orphan nuclear receptor RORγt and interleukin(IL) -17.Serum IL-6 and IL-23 were detected by ELISA method.Results ( 1 ) Compared with healthy controls,the frequency of Th17 cells [ (0.75± 0.79) % vs ( 0.28 ± 0.23 )% ] and expression of RORγt ( 0.30 ± 0.38 vs 0.04 ± 0.02,both P < 0.05 ) were significantly increased in HT patients.( 2 ) The levels of IL-6 and IL23 in HT patients were higher than those in healthy controls ( 3.66 ± 4.70 vs 0.47 ± 1.11,154.7 ± 75.81 vs 80.65 ± 61.41,both P<0.05 ).( 3 ) A positive correlation between Th17 cells and serum TgAb was revealed in HT patients ( r =0.848 4,P =0.007 7 ).(4) The results of PCR showed that IL-17 and RORγt expressed in thyroid tissues of patients HT.Conclusion An increased frequency of Thl7 cells was found in HT patients,implying that this cell subset may play an important role in the pathogenesis of autoimmune thyroid disease.
3.Study on the value of T-SPOT.TB test in tuberculous pleurisy
Liang GAO ; Jian ZHENG ; Qinfang OU ; Huaxi CHEN ; Zhiming YU ; Yanbin CHEN
The Journal of Practical Medicine 2015;(17):2833-2835
Objective To explore the value of T-SPOT.TB test in diagnosis of tuberculous pleurisy by comparing the T-SPOT.TB test, adenosine deaminase (ADA) of hydrothorax and tuberculosis antibody (TB-Ab). Methods 62 pleural effusion patients are included in the research , of which 32 cases have tuberculosis and 30 cases have no tuberculosis. All patients underwent T-SPOT.TB, pleural effusion ADA, and TB-Ab test. The results were compared with final clinical diagnosis for sensitivity and specificity evaluation. Results The sensitivity of T-SPOT.TB, ADA, TB-Ab were 90.6%, 71.9% and 62.5% respectively. The specificity of T-SPOT. TB, ADA, TB-Ab were 90.0%, 83.3% and 86.7% respectively. The sensitivity of T-SPOT.TB was the highest one among the three methods. The sensitivity of T-SPOT.TB has statistically significant difference compared with TB-Ab (P < 0.05), but no statistically significant difference compared with ADA (P > 0.05). Conclusions The T-SPOT.TB test had higher sensitivity and specificity for the diagnosis of tuberculous pleurisy , and had important reference value in early diagnosis of patients with tuberculous pleurisy. The T-SPOT.TB and TB-Ab combination examination had a complementary effect.
4.Role of interleukin-17 in spinal dorsal horns in neuropathic pain in rats and its effect on activation of astrocytes
Caixia SUN ; Wen YUAN ; Chunye LI ; Yanfang LIU ; Pan CHEN ; Zhaoliang SU ; Huaxi XU
Chinese Journal of Anesthesiology 2015;35(3):320-325
Objective To investigate the role of interleukin-17 (IL-17) in spinal dorsal horns in neuropathic pain (NP) in rats and its effect on activation of astrocytes.Methods In vivo experiment Sixty-four male SPF Sprague-Dawley rats,aged 6-8 weeks,weighing 180-200 g,were randomly divided into 3 groups using a random number table:control group (group C,n =16),sham operation group (group S,n =24) and group NP (n =24).The animals were anesthetized with intraperitoneal pentobarbital sodium,the L5,6 spinal nerves of the left side of the rat were gently separated and exposed,tightly ligated with 5-0 silk suture and transected.In group S,the L5,6 spinal nerves of the left side of the rat were only exposed.In group C,no operation was performed.Mechanical pain threshold was measured at day 1 before operation and days 1,3,5,7,10 and 14 after operation.The expression of IL-17,IL-6,IL-1β and tumor necrosis factor-alpha (TNF-α) mRNA in the spinal dorsal horn was determined using quantitative real-time PCR at day 7 and day 14 after operation.At day 7 after operation,the activation of astrocytes in the spinal dorsal horn was detected.In vitro experiment Primarily cultured astrocytes of neonatal rats were randomly divided into 4 groups using a random number table:control group (group C,n=22),10 ng/ml IL-17 group (I10 group,n=18),50 ng/ml IL-17 group (I50 group,n-18) and 100 ng/ml IL-17 group (I100 group,n=22).In I10,I50 and I100 groups,the astrocytes were incubated with the culture medium containing 10,50 and 100 ng/ml IL-17,respectively.The proliferation of astrocytes was detected by MTT at 24,48 and 72 h of incutation or culture.The expression of IL-6,IL-1β and TNF-α mRNA was determined using quantitative real-time PCR.Results In vivo experiment Compared with group C,the mechanical pain threshold was significantly decreased at 3-14 days after operation,the expression of IL-17,IL-6 and IL-1β mRNA in the spinal dorsal horn was up-regualted at 7 days after operation,and the activation of astrocytes was increased in group NP,and no significant change was detected in the mechanical pain threshold at each time point after operation in group S.In vitro experiment Compared with group C,the proliferation of astrocytes was significantly increased at 48 h of incubation in I10 and I50 groups,the proliferation of astrocytes was significantly increased at 48 and 72 h of incubation,and the expression of IL-6 and IL-1β mRNA was up-regulated in I100 group,and no significant change was found in the proliferation of astrocytes in group S.Conclusion Up-regulated expression of IL-17 in spinal dorsal horns may be involved in the maintenance of NP,and the mechanism is related to promoted activation of astrocytes and induced inflammatory responses in rats.
5.Preparation and characterization of transfersomes of three drugs in vitro.
Yu ZHENG ; Shi-xiang HOU ; Tong CHEN ; Yi LU
China Journal of Chinese Materia Medica 2006;31(9):728-731
OBJECTIVETo investigate the influence of drug properties on the encapsulation effiency (EE) and drug release of transfersomes for a proper transfersome preparation.
METHODTo prepare the transfersomes of colchicines (CLC), vincristine sulfate (VCR) and mitoxantrone hydrochloride (DHAD) with the same materials and methods, and then measure their EE. To find out the relationship between drug properties like solubility, molecular weight and charges, and EE. To performe the drug release experiments of various types of transfersomes in vitro, and compare their differences.
RESULTVCR and DHAD are lipophilic or hydrophilic, owing positive charges and large molecular weight, as a result, their EE are high, while CLC is amphipathic, neutral, and of small molecular weight, its EE is very low. As DHAD can insert into the membrane of transfersome, the drug release of DHAD-T in vitro is much slower than that of VCR-T.
CONCLUSIONTo prepare transfersomes with high EE, drugs that are lipophilic or hydrophilic, high molecular weight and opposite charges to the membrane should be chosen. Interaction between drugs and membrane will influnce the rate of drug release.
Antineoplastic Agents ; administration & dosage ; chemistry ; Antineoplastic Agents, Phytogenic ; administration & dosage ; chemistry ; Colchicine ; administration & dosage ; chemistry ; Deoxycholic Acid ; Drug Carriers ; Gout Suppressants ; administration & dosage ; chemistry ; Mitoxantrone ; administration & dosage ; chemistry ; Particle Size ; Phosphatidylcholines ; Solubility ; Technology, Pharmaceutical ; methods ; Vincristine ; administration & dosage ; chemistry
6.IL-10 producing regulatory B cells ameliorated cardiac hypertrophy of viral myocarditis mice
Shiqing ZHANG ; Jia CHEN ; Lin XIA ; Zhaoliang SU ; Huaxi XU ; Fang LIU
Chinese Journal of Microbiology and Immunology 2021;41(5):345-352
Objective:To explore the mechanism of B10 cell involved in cardiomyocyte hypertrophy following myocarditis, and to develop potential therapeutic strategies.Methods:BALB/c mice infected with Coxsackie virus B3 induced viral myocarditis model. The expression of angiotensin (ANG)Ⅱ and its receptor in myocarditis mice was detected. The changes of B10 cells in the hearts of control mice and myocarditis mice were analyzed by flow cytometry. After losartan was administered to myocarditis mice, the degree of myocardial inflammation was detected by HE staining, the expression of inflammatory factors was detected by ELISA, the myocardial hypertrophy was detected by wheat germ agglutinin (WGA) staining, and the changes of B10 cells in the heart were analyzed by flow cytometry. The levels of cardiac troponin T (C-TNT) and high mobility group box 1 (HMGB1) protein in neonatal mouse cardiomyocytes treated with ANGⅡ and ANGⅡ+ IL-10 were detected. Cardiomyocytes were treated with ANGⅡ, ANGⅡ+ B10 cells, ANGⅡ+ B10 cells + IL-10 receptor antibody and ANGⅡ+ B cells to detect C-TNT protein levels, and Annexin-V/PI was used to detect the apoptosis of cardiomyocytes. Cardiomyocytes were treated with oxidized HMGB1, reduced HMGB1 and disulfide HMGB1, and C-TNT expression was detected.Results:Coxsackievirus B3 infection caused cardiac hypertrophy, high expression of ANGⅡ and its receptor, and transient increase of B10 cells in mice. Losartan treatment blocked the angiotensin receptor, reduced expansion of B10 cells. B10 cells alleviated apoptosis of cardiomyocytes and inhibited the production of HMGB1 induced by ANGⅡ patch by producing IL-10, thus alleviating viral myocarditis and cardiac hypertrophy.Conclusions:B10 cells may play an important role in myocardial protection in myocarditis.
7.Studies on purification of the extract of fructus Gardeniae for injection by macroreticular resins.
Gang CHEN ; Shi-xiang HOU ; Li-min YE ; Ping HU
China Journal of Chinese Materia Medica 2005;30(3):188-190
OBJECTIVETo purify the extract of fructus gardeniae for injection by macroreticular resins in purification process of traditional Chinese medicine (TCM) Injections.
METHODUsing fructus gardeniae as sample, on base of obtaining the extract by employing macroreticular resin, quality evaluation and rationality of purification methods had been studied by the quantitative analysis of active ingredients and the characteristics of micromeritics, safety and stability of the extract.
RESULTThe experiment showed the extract of fructus gardeniae for injection had been produced successfully by macroreticular resin.
CONCLUSIONUsing macroreticular resins is a promising purification way of TCM injections, whereas a more consummate method of quality evaluation must be established to ensure safety, efficiency and stability of the preparation in the process.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Fruit ; chemistry ; Gardenia ; chemistry ; Injections ; Iridoids ; isolation & purification ; Mice ; Plants, Medicinal ; chemistry ; Pyrans ; isolation & purification ; Quality Control ; Resins, Synthetic ; Technology, Pharmaceutical ; methods
8.Acupuncture for the lower extremity spasm induced by spinal injury: random controlled trial of single case.
Chinese Acupuncture & Moxibustion 2010;30(6):473-477
OBJECTIVETo study whether acupuncture on the acupoints of the affected extremities was effective or not on the lower extremity spasm in spinal injury, and observe the tolerance of patients to acupuncture.
METHODSThe random controlled trial of single case was adopted among 9 patients. The heavy manipulation and gentle manipulation of acupuncture on the lower extremities, physical therapy as well as corresponding control therapy (acupuncture or other therapy on the upper extremities) were applied to each case separately for 3 cycles of treatment, totally for 29 weeks of treatments. The therapeutic effective outcomes were determined according to the index of clinical spasm, daily spasm frequency and the patient reported outcome.
RESULTSNone of the three therapies made the index of clinical spasm worse in 9 patients (all P > 0.05), and there was no significant statistical difference in comparison among three therapies (P > 0.05). Concerning to the improvement in daily spasm frequency, the result of heavy manipulation therapy was inferior apparently to the other two therapies (P < 0.05). The outcome report indicated that the patients were unwilling to accept the stimulation induced by heavy manipulation.
CONCLUSIONThe spasm induced by spinal injury can not be aggravated in tendency by acupuncture on the affected extremities. Thus, acupuncture is a kind of safe therapy for spasm. But the heavy manipulation of acupuncture may be the factor to impact the patient for the option of acupuncture treatment.
Acupuncture Points ; Acupuncture Therapy ; Adult ; Humans ; Lower Extremity ; injuries ; Male ; Middle Aged ; Spasm ; etiology ; therapy ; Spinal Cord Injuries ; complications
9.Expression of telomerase activity, telomerase RNA component and telomerase catalytic subunit gene in lung cancer.
Weiwei CHEN ; Xiaoxiong XIONG ; Hongyuan ZHOU ; Qinghua ZHOU
Chinese Medical Journal 2002;115(2):290-292
OBJECTIVETo investigate whether telomerase activity, human telomerase RNA (HTR) and human telomerase reverse transcriptase (HTERT) expression were associated with tumor development in lung cancer and whether telomerase is regulated at gene level or transcriptional level.
METHODSExpression of HTR and HTERT was detected by reverse transcription-polymerase chain reaction (RT-PCR) in 68 human lung cancer and in 68 adjacent-neoplatic lung tissues. And telomerase activity was examined by a quantitative telomeric repeat amplification protocol (TRAP).
RESULTSIn 68 lung cancer tissues, telomerase activity, HTR and HTERT were expressed in 79%, 98.5% and 91.2% respectively, whereas all adjacent non-neoplastic lung tissues were telomerase negative. Most normal lung tissues expressed HTR (91.2%) and HTERT was detected in only 7 of 68 non-neoplastic tissues.
CONCLUSIONSThe relatively high frequency of telomerase activity in lung cancer whereas the detection of no telomerase activity in normal lung tissues suggested that telomerase may play an important role in tumorigenesis of lung cancer. Compared to HTR, HTERT expression was better associated with telomerase activity with a concordance of 88.9%. Telomerase activity may be regulated at transcription level or translation level.
DNA-Binding Proteins ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; enzymology ; genetics ; RNA ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; genetics
10.Assessment of risk for tin mine dust in Guangxi.
Huasheng ZHANG ; Jian YANG ; Yingbiao QIN ; Rangan CHEN ; Jingqiong CHEN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2002;20(6):430-432
OBJECTIVETo explore the minimum observed adverse effect level(LOAEL) and intensity of pathogenesis of tin mine dust.
METHODSA cohort study design with retrospective assessment of exposure was used. Selected 4,471 male tin miners who were exposed to tin at least one year during 1960 to 1974 and were compared with 4,797 pottery dust exposed workers in the same way designed. Statistical analysis system SAS, PROC LIFETEST were used to perform the non-parameter calculation by Life Table method.
RESULTSTin mine cohort subjects were followed up to December 31, 1994. The percentage of miners who developed silicosis was 21.7% (971/4,471). 81% of the patients had been exposed to dust before 1958. The cumulative total dust exposure(CTD) was significantly correlated with silicosis risk. The risk of silicosis was 0.012 when CTD was less than 50 mg/m-3.year-1. The risk of silicosis was increased to 0.971 when CTD was beyond 400 mg.m-3.year-1. However, the risk of silicosis was only 0.369 for dust exposed workers in pottery factories when CTD was beyond 400 mg.m-3.year-1.
CONCLUSIONThere was significant relationship between cumulative dust exposure and the incidence of silicosis in tin exposed workers. And silicosis induced by tin mine dust is more serious than the pottery dust.
Cohort Studies ; Dust ; Humans ; Mining ; Occupational Exposure ; adverse effects ; Silicosis ; etiology ; Tin