1.Expression of a Novo Human Osteoprotegerin-Mycobacterial Heat Shock 70 Fusion Protein with Baculovirus-Insect Expression System
Shu LIU ; Quan-Geng ZHANG ; Yue ZHANG ; Jing MA ; Zhen-Long LIU ; Wen-Ming ZHAO ;
China Biotechnology 2006;0(05):-
Objective:Osteoprotegerin (OPG) is a key molecule negatively regulating osteoclast differentiation and activation; and the conserved mycobacterial heat shock 70 (HSP70) peptide p111-125 has also been found to inhibit inflammation reactions in chronic arthritis. BaculoDirectTM baculovirus expression system was selected to express recombinant OPG-HSP70 in insect cells.Methods:The human functional fragment (p22-194) of OPG and functional fragment (p111-125) of mycobacterial HSP70 gene were cloned into the transfer vector pENTRTM/SD/D-TOPO. The recombinant plasmid was performed an LR reaction with the BaculoDirectTM Linear DNA to generate recombinant baculovirus DNA. The cultured Sf9 insect cells were directly transferred with the recombinant baculovirus DNA,and the pure recombinant baculovirus was obtained. Then recombinant baculovirus was infected Sf9 insect cells again to express the OPG-HSP70 gene.Results:The target protein was detected at the time of 48h post infection,reached at highest yield at the time of 72h post-infection. A 28kDa protein immunostaining band was detected by Western blotting from lysate of those cells.And the purified protein was obtained by using Ni-NTA system. Functional stuies on the fusion protein showed it significantly reduce osteoclast cell number[(3.10?0.640) cells under each microscope field in treatment group by comparing to (10.70?0.817)cells in the control group] in the osteoclast inhibition test,and reduce the inflammation reaction in a delayed type hypersensitivity (DTH) mice model (P
2.Analysis of iodine nutrition of vulnerable population in the high risk region of iodine deficiency disorders in Qinghai province in 2009
Li-lin, CHEN ; Duo-long, HE ; Shu-bang, LI ; Xun, CHEN ; Zhi-jun, ZHAO ; Qiang, ZHANG
Chinese Journal of Endemiology 2011;30(5):539-541
Objective To investigate the iodine nutrition level of the vulnerable population in the high risk region of iodine deficiency disorders for providing a scientific basis for the development of effective preventive measures in Qinghai province. Methods The non-iodized salt coverage rate > 5% of Delingha city, the coverage rate of iodized salt and the rate of qualified iodized salt ≤80% of Jiuzhi, Wulan, Nangqian, Zaduo, Geermu,Yushu, Gande, Chengduo, and Dulan counties(cities), the median of urinary iodine < 100 μg/L of Huangyuan county of 11 areas were chosen as monitoring area in 2009. Five townships(towns) were selected by their location of east, west, south, north and center in each county (city), and one school was selected in each township (town),and 60 household salt samples were collected in each school, and quantitatively determined by direct titration of iodine(GB/T 13025.7-1999). Three townships(towns) were selected in each county(city), and two schools were selected in each township(town). The content of urinary iodine of 40 children aged 8 - 10 and 20 women of childbearing age aged 18 - 40 was analyzed by As3+-Ce4+ catalytic spectrophotometry (WS/T 107-2006). ResultsThree thousand two hundred and sixty-one edible salt samples were tested. The coverage rate of iodized salt was 79.88%.The iodized salt qualification rate was 95.16%. The qualified iodized salt consumption rate was 76.02%. The noniodized salt coverage rate was 20.12%. In Nangqian, Zaduo, Yushu and Geermu counties(cities), the non-iodized salt coverage rate was 88.89%, 45.05%, 43.00% and 12.67%, respectively. The median of urine iodine of 2536 children aged 8 - 10 was 155.8 μg/L, with ≤50 μg/L about 13.6% (346/2536), > 100 μg/L about 67.5%(1711/2536), and it was 75.1, 94.6 μg/L in Nangqian and Zaduo counties, respectively. The median of urine iodine of 665 women of childbearing age was 129.7 μg/L, with ≤50 μg/L about 22.7%(151/665), > 100 μg/L about 59.2%(394/665), and it was 21.0, 54.7, 72.7 μg/L in Zaduo, Nangqian and Chengduo counties, respectively.Conclusions Insufficient intake of iodine exits in children and women of childbearing age in high risk region of iodine deficiency disorders in Qinghai province, which should be corrected as soon as possible.
3.Advances in research and development of universal influenza vaccines.
Li-Xia ZHANG ; Jian-Fang ZHOU ; Yue-Long SHU ; Bao-Shou YANG ; Zhao-Qing HE
Chinese Journal of Virology 2014;30(1):73-78
Vaccination is the primary strategy for the prevention and control of pandemic influenza. Because influenza virus is highly variable across strains, universal influenza vaccines need to be developed to address this problem. This review describes the research progress in conserved epitopes of influenza virus, the advances in the research and development of universal influenza vaccines based on the relatively conserved sequences of NP, M2e, HA2, and headless HA, the mechanisms of cross-protection, and the methods to improve cross-protection.
Animals
;
Cross Reactions
;
Humans
;
Orthomyxoviridae
;
immunology
;
Species Specificity
;
Viral Proteins
;
immunology
;
Viral Vaccines
;
genetics
;
immunology
4.Clinical observation on repair of lymphocyte injury in patients with diabetic nephropathy treated by regulating spleen-stomach needling.
Zhi-Long ZHANG ; Shu-Hua ZHAO ; Xin LI ; Yuan-Qing YANG ; Hong CHEN ; Man WANG
Chinese Acupuncture & Moxibustion 2013;33(12):1065-1070
OBJECTIVETo explore therapeutic effect and action mechanism of regulating spleen-stomach needling on diabetic nephropathy (DN).
METHODSUsing multi-centric, randomized, controlled and blind principles, 144 cases of DN were divided into an observation group and a control group according to random digital tab, 72 cases in each one. Based on regular treatment of diabetes, the regulating spleen-stomach needling was applied at Zhongwan (CV 12), Quchi (LI 11), Hegu (LI 4) and Xuehai (SP 10), etc. in the observation group while Shenshu (BL 23), Taixi (KI 3), Sanyinjiao (SP 6), Yanglingquan (GB 34), etc. were selected in the control group by reference of Acupuncture and moxibustion. The treatment was given twice a day, six days as a treatment session with interval of one day between sessions. Totally six weeks were required. Changes of clinical symptoms and signs, fast blood glucose (FBG), urinary albumin excretion rate (UAER), beta2-microglobulin (beta2-MG), monocyte chemotactic protein-1 (MCP-1), lymphocyte membrane cholesterol, propanediol (MDA), PCO, 8-hydroxydeoxy guanosine (8-OHdG), superoxide dismutase (SOD), CD3+, CD4+, CD8+, and CD4+/CD8+ were observed before and after treatment in two groups.
RESULTSAs for improving clinical symptoms and signs, total effective rate was 84.29% (59/70) in the observation group and 55.56% (40/72) in the control group, which had statistical difference between two groups (P<0.01). As for regulating glycometabolism [(6.25 +/- 0.32) mmol/L vs (8.09 +/- 0.63) mmol/L], reducing UAER [(154.43 +/- 55.14) mg/24h vs (268.91 +/- 77.65) mg/24h], restraining over-expression of MCP-1 [(137.59 +/- 36.15) pg/mL vs (166.89 +/- 42.82) pg/mL], regulating level of oxidative stress, prohibiting oxidation of protein and adjusting quantity and activity of T lymphocyte subgroup, the observation group was superior to the control group (P< 0.05, P<0.01).
CONCLUSIONThe regulating spleen-stomach needling is an effective method for treatment of DN, which cold improve glycometabolism disturbance-induced progressive kidney injury, recover glomerular filtration, reduce urinary albumin excretion rate, restrain overexpression of MCP-1, adjust level of oxidative stress, prohibit oxidation of protein, increase protectiveness of membrane, adjust quantity and activity abnormity of T lymphocyte subgroup, leading to repairing lymphocyte damage and improving immune expression to delay kidney damage.
Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Diabetic Nephropathies ; immunology ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Spleen ; physiopathology ; Stomach ; physiopathology
5.Susceptibility of 15 collections of Aedes albopictus from Guizhou to dengue virus oral infection.
Li-ping SHU ; Li ZUO ; Xing ZHAO ; A-ying CHEN ; Long-hua WEI
Chinese Journal of Experimental and Clinical Virology 2004;18(3):234-237
OBJECTIVETo study the susceptibility of Aedes albopictus to dengue virus infection.
METHODSAedes albopictus from 15 collections in Guizhou province were challenged orally with dengue virus 1-4 types, respectively. The total RNA from mosquitos were extracted. The viral NS1 gene fragment was amplified with reverse transcriptase polymerase chain reaction (RT-PCR). Dengue virus in mosquitoes was isolated with C6/36 cells. Then the viral antigen was detected by indirect immunofluorescence assay (IFA). Antigen and nucleic acid of dengue virus from 15 geographic strains of Aedes albopictus orally infected with dengue viruses (DEN-1, DEN-2, DEN-3 and DEN-4) were detected by IFA, RT-PCR and the virus was isolated with C6/36 cells, respectively.
RESULTSThe rates of Aedes albopictus orally infected with DEN-1, DEN-2, DEN-3 and DEN-4 were 12/15, 12/15, 8/15 and 13/15, respectively.
CONCLUSIONSDifferent geographic strains of Aedes albopitus in Guizhou were susceptible to dengue viruses.
Aedes ; cytology ; virology ; Animals ; Antigens, Viral ; analysis ; Cell Line ; China ; DNA, Viral ; analysis ; Dengue Virus ; genetics ; isolation & purification ; Disease Susceptibility ; Ecosystem ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Nonstructural Proteins ; genetics
6.Histopathologic characteristics of intestinal metaplasia in gastric mucosa of children.
Gui-ping CHEN ; Hong-feng TANG ; Wei-zhong GU ; Hua-ying YE ; Long LIN ; Yan SHU ; Yun ZHAO
Chinese Journal of Pathology 2006;35(3):171-172
Adolescent
;
Child
;
Child, Preschool
;
Female
;
Gastric Mucosa
;
pathology
;
Gastritis
;
pathology
;
Humans
;
Male
;
Metaplasia
;
Retrospective Studies
7.A study on the long-term outcome of hepatitis B e antigen-negative chronic hepatitis B compared with that of hepatitis B e antigen-positive chronic hepatitis B
Gui-Cheng WU ; Wei-Ping ZHOU ; You-Tong ZHAO ; Shu-Hua GUO ; Ai-Long HUANG ; Hong REN ; Ding-Feng ZHANG ;
Chinese Journal of Infectious Diseases 2001;0(03):-
0.05)between the two groups.Conclu- sion The long-term outcomes of e-CHB is not markedly different compared with that of e+CHB.
8.Ultrasound-induced uterine smooth muscle contraction
Shu-Fang CHANG ; Shen-Yin ZHU ; Zhi-Biao WANG ; Kai HU ; Zhi-Long WANG ; Chun-Liang ZHAO ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(07):-
Objective To study the influence and dose effect of ultrasound on the contraction of uterine smooth muscle in rats.Methods Estradiol benzoate was injected into rats three days before conducting an in-vitro experiment.Their uteri were resected and irradiated with ultrasound(0.8 MHz,3 W/cm~2,0-40 rain).The contrac- tion frequency and amplitude were recorded using an MS-302 biological experiment system.Results It could be seen that the contraction frequency and amplitude,and general contractile activity were significantly increased during ultrasonic irradiation(P<0.01).The increased contraction frequency and amplitude lasted for ten minutes,and then the normal contraction pattern resumed.The contraction frequency as well as the percentage change in eontraction fre- quency were highest during the first 15 minutes of ultrasonic irradiation;the contraction amplitude as well as the per- centage change in amplitude were highest during 40 minutes of ultrasonic irradiation.Contraction activity was at its highest for 30 minutes,but the percentage change in activity was highest for 20 minutes.Conclusions Ultrasound can induce uterine smooth muscle contraction in rats.This biological effect is related to the irradiation time.
9.Comparative study of myocardial perfusion imaging and 64 multi-slice spiral CT for the diagnosis of coronary artery disease
Jun, ZHAO ; Long-bao, XU ; Ren-ming, WAN ; Guang-lei, FAN ; Jian-wen, LIU ; Shu-xing, HUANG
Chinese Journal of Nuclear Medicine 2010;30(6):367-371
Objective To compare the diagnostic value of myocardial perfusion imaging (MPI) and 64 multi-slice spiral CT (64-MSCT) for coronary artery disease (CAD). Methods Fifty-two patients with suspected or known CAD were included in the study. Each patient underwent both stress and rest MPI,MSCT as well as conventional coronary angiography (CAG) within 1 month. The stress and rest MPI were scored by a 5-grade criteria (0 ~ 4) based on 17 coronary artery segments. The difference between summed stress and rest scores > 1 was defined as myocardial ischemia. Stenosis in one main vessel or one main branch of the main vessel ≥50% was defined as myocardial ischemia by MSCT. CAG was used as the reference for comparison. Statistical analysis was performed using SPSS 13. 0 software. Kappa value was used to test the accordance of MPI and MSCT results. X2 test was used to evaluate the difference between MPI and MSCT results. Results The patient-based sensitivity, specificity, positive and negative predictive values and accuracy of MPI and MSCT for the diagnosis of CAD were 86.7% (26/30), 77.3% ( 17/22),83.9% (26/31), 81.0% ( 17/21), 82.7% (43/52) and 83.3% ( 25/30), 86.4% ( 19/22), 89.3%( 25/28), 79.2% ( 19/24), 84.6% (44/52), respectively. The vessel-based sensitivity, specificity, positive and negative predictive values and accuracy of MPI and MSCT were 74.5% (38/51), 81.0% (85/105 ), 65.5% (38/58), 86.7% ( 85/98), 78.8% ( 123/156 ) and 90.2% (46/51 ), 88.6% ( 93/105 ),79.3 % (46/58), 94.9% (93/98), 89.1% ( 139/156), respectively. There was no statistically significant difference between MPI and MSCT for either patient or lesion-based diagnosis (X2 =0.44, 0.21, both P >0.05 ). 96.0% (24/25) patients with both abnormal MPI and MSCT positive were valified by CAG while 83.3% (15/18) patients with both MPI and MSCT negative were excluded by CAG. Conclusions Both MPI and MSCT are reliable diagnostic modalities for CAD. They also provide complementary diagnostic value to each other.
10.Relationship between maternal and neonatal thyroid function in a high iodine area
Wei, WEI ; Hua, LIU ; Shu-fen, ZHANG ; Zhong-na, SANG ; Gui-qin, ZHANG ; Na, ZHAO ; Long, TAN ; Wan-qi, ZHANG
Chinese Journal of Endemiology 2012;31(1):37-39
Objective To find out iodine nutritional status of pregnant women in the third trimester and the relationship between pregnant women and their neonates thyroid function in a high iodine area.Methods From April to June 2010,210 copies of fasting morning urine and venous blood,and their neonatal umbilical cord blood samples were collected in Haixing Hospital,Cangzhou city,Hebei province.Urinary iodine was determined by arsenic-cerium catalytic spectrophotometry.The levels of free triiodothyronine(FT3),free thyroxine (FT4) and sensitive thyroid-stimulating hormone (sTSH) in serum were measured by chemiluminescence.Results Median urinary iodine of 210 pregnant women(average age 27.69 ± 4.73 years) in the third trimester was 1240.70 μg/L,84.3%(177/210) of them was in excessive iodine nutrition,and only 0.5%(1/210) of them was in adequate iodine nutrition.The prevalence rate of thyroid diseases was 19.5% (41/210),and the spectrum of diseases were subclinical hypothyroidism(16.2%,34/210),subclinical hyperthyroidism(0.9%,2/210),hypothyroidism(2.4%,5/210) and hyperthyroidism (0,0/210).The number of newborns with sTSH 5 - > 10 mU/L were 104 persons (49.5%); 10 - > 20 mU/L were 44 persons(21.0%),and ≥20 mU/L were 16 persons(7.6%).Of pregnant women suffer from thyroid disease,the ratio(50.0%,24/48 ) of sTSH equal to 10.18 mU/L and > 10 mU/L of their neonates was higher than that of their corresponding non-ill pregnant women(6.78 mU/L,Z =- 2.867,P < 0.05; 22.2%,36/162,x2 =14.000,P < 0.05).There was a positive correlation between neonates' and their mothers' sTSH levels (r =0.278,P < 0.05).There was also a positive correlation between neonates' (sTSH > 10 mU/L) and their mothers' abnormal sTSH levels (r =0.240,P < 0.05).Conclusions Most of the pregnant women in high iodine areas are iodine excess.The level of neonates' sTSH is higher,and it is due to their mothers' abnormal sTSH and suffering from thyroid diseases to some extent.As a result,the monitoring of pregnant women's iodine nutrition and thyroid function and sTSH level of their neonates should be strengthened.