OBJECTIVETo study the effects of zuogui pill (ZP) contained serum on the gene expressions of type-II collagen and proteoglycan during the differentiation of mesenchymal stem cells (MSCs) towards chondrocytes.

METHODSMSCs isolated from rat bone marrow were in vitro induced differentiation towards chondrocytes and stimulated with high- (57 g/kg), middle- (28.5 g/kg), and low-dose (9.5 g/kg) ZP contained serums and serum of blank rats. The proliferation of MSCs was analyzed by CCK-8 method. The 3rd-passage MSCs were divided into the blank control group (by adding serum of the blank group rats), the induction control group (by adding the induction fluid and serum of the blank group rats), and the ZP contained serum group (by adding the induction fluid and middle-dose ZP contained serum). The expressions of type-II collagen and proteoglycan were determined using reverse transcriptase PCR, Real-time PCR, and immunohistochemistry.

RESULTSCompared with the blank control group, the proliferation of MSCs could be promoted by ZP contained serum at different doses (P < 0.05), with the most obvious effect shown in the middle-dose ZP contained serum group (P < 0.05). The mRNA and protein expressions of type-II collagen could be identified in the induction control group and the ZP contained serum group on the 21st day of the induction. Of them, the mRNA expression of type-II collagen in ZP contained serum groups was obviously higher than that of the induction control group. Results of Real-time PCR showed that on the 21st day of the induction, the mRNA expression quantitation of proteoglycan in ZP contained serum groups was about 16-fold and 3-fold of the levels on the 7th day and the 14th day (P < 0.05), obviously higher than those of the induction control group (P < 0.05).

CONCLUSIONZP contained serum could induce MSCs proliferation, the gene expressions of type- II collagen and proteoglycan, which might be one of its molecular bases for protecting the cartilage.

Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; metabolism ; Collagen Type II ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Proteoglycans ; metabolism ; Rats

OBJECTIVETo study influenza epidemic and analyze antigenic and genetic characterization of the predominant strains in Wuhan area in 2003.

METHODSEpidemiological data and specimens from influenza patients were collected from surveillance sites weekly. Viruses were isolated from the specimens. Three H3 isolates were chosen to do antigenic analysis by hemagglutination inhibition (HI) test and their HA1 region was sequenced.

RESULTSTotally 58 influenza viruses were isolated from 418 specimens, 57 of them were identified as H3 subtype and 1 of them was B subtype; both monthly positive rate and numbers of influenza like illness had two peaks of winter and summer, the highest peak appeared in July. The 3 new H3 isolates were antigenically different from vaccine strain A/Panama/2007/99, 14 amino acid changes have been found in HA1 domain of these 3 strains compared with A/Panama/2007/99, phylogenetic analysis also confirmed the difference in HA1 domain.

CONCLUSIONSInfluenza epidemic had two peaks in Wuhan area in 2003. The activity of H3 virus was strengthened remarkably. And they are antigenically and genetically different from the vaccine strain.

Amino Acid Sequence ; Antigens, Viral ; immunology ; China ; epidemiology ; Genes, Viral ; Glycosylation ; Hemagglutination Inhibition Tests ; Humans ; Influenza A Virus, H3N2 Subtype ; genetics ; immunology ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, Protein

A group of 25 rabies viruses (RABVs),recovered from 24 dogs and one human case,were collected from various areas in China between 2004 and 2006.Genetic and phylogenetic analyses of the G-L intergenic region were carried out in 25 street RABV isolates and CTN vaccine strains of 7 generations.The study was based on the comparison of a 519 bp nucleotide sequence,encompassing the G-L intergenic region.The nucleotide sequence homologies of Chinese street strains were from 95.5% to 100%.The phylogenetic analysis showed that all Chinese isolates clearly supported the placement of all Chinese viruses in Lyssavirus genotype 1 and they were distributed according to their geographical origins.All of the Chinese strains were closely related but they could still be divided into two groups:group of street strains and group of CTN strains.This study presents details about the molecular epidemiology of rabies viruses based on the sequences of the G-L Intergenic region.

Objective: To explore the association between chronic hepatitis B virus infection and diabetes among adults. Methods: The baseline data of China Kadoorie Biobank ( CKB ) study from Tongxiang of Zhejiang Province was used for analysis. Community residents were investigated in the study from August 2004 to May 2008, including questionnaire survey, physical measurement and biological sample test. Univariate and multivariate logistic regression models were used to estimate the association of chronic hepatitis B virus infection with diabetes. Results: Totally 52 888 participants were included in the final analysis. The overall prevalence of HBsAg-positive was 3.55% ( N=1 877 ). The overall prevalence of diabetes was 5.17% ( N=2 733 ). The prevalence of HBsAg-positive in diabetic patients was 3.51% ( N=96 ). Both univariate and multivariate logistic regression models indicated that there was no association between chronic hepatitis B virus infection and diabetes（ P>0.05 ）. Conclusion No significant association has been found between chronic hepatitis B virus infection and diabetes among adults.

AIMTo synthesize and identify artificial antigen of podophyllotoxin for the production of podophyllotoxin polyclonal antibody.

METHODSThe hapten was synthesized by two different chemical approaches and characterized by TLC, IR, NMR, and MS. Mixed anhydride reaction (MAR) and active ester method (AEM) were used to couple the podophyllotoxin to carrier proteins (BSA and OVA). Characterization of artificial antigens was done by using spectroscopy and electrophoresis. The anti-podophyllotoxin polyclonal antibodies were obtained through immunizing rabbits.

RESULTSThe results from IR, NMR and MS showed that 4-O-succinoyl podophyllotoxin (hapten) was successfully synthesized. The coupling molar ratios of the hapten and carrier proteins were 88.6 for Hapten-BSA1, 40.3 for Hapten-BSA2, 17.8 for Hapten-OVA1, and 54.2 for Hapten-OVA2. Hapten conjugates coupled with BSA yielded two sets of the specific and affinitive polyclonal antibodies. One set of antibodies showed an IC50 value of 2.21 microg.mL(-1) with a detection limit of 0.12 microg.mL(-1).

CONCLUSIONAntigenic conjugates were artificially synthesized, and based on these artificial antigens, polyclonal antibodies against podophyllotoxin were raised from rabbits immunized with two different immunogens and characterized with an indirect ELISA format.

Animals ; Antibodies ; analysis ; Antibody Affinity ; Antibody Formation ; Antineoplastic Agents, Phytogenic ; immunology ; Enzyme-Linked Immunosorbent Assay ; Haptens ; chemistry ; immunology ; Immune Sera ; chemistry ; Male ; Ovalbumin ; immunology ; Podophyllotoxin ; immunology ; Proteins ; immunology ; Rabbits ; Serum Albumin, Bovine ; immunology

For epidemiological investigation of the rabies virus carrier rates of domestic dogs, cats and wild animals like rodent animals and bats,three kinds of regions where rabies had higher incidence (Hunan and Guizhou Provinces), lower incidence (Jiangsu Province, Wuhan City) and provisionally rabies-free (Shenyang City) were selected. Then the antigenic types, the genovariation of the isolaled viruses and the currently vaccine matching of the virus strains were analyzed. The results showed that in China the principal host of rabies is dog,the total virus carrier rate of the captured dogs was 2.56%, and the highest positive isolation rate was 20.0% in some monitoring site. However,there was no evidence about the rabies virus carrier rate in rodent animals,bats or other wild animals. The rabies vaccines which prepared from aG and CTN strains have already been produced successfully in China. The research showed that the nucleotide sequences of the newly isolated viruses were more similar with the glycoprotein gene of CTN strain. In order to evaluate the safety and the efficacy of the vaccines currently used, two groups (50 people each) were injected with vaccine of aG strain and CTN strain respectively in five surveillance points. The neutralizing antibody tested were 0.49 IU/mL-0.52 IU/mL and 6.7 IU/mL-7.53 IU/mL after the 7 and the 14 days of vaccine injection respectively. In addition, the rates of antibody positive seroconversion were 45.1%-47.9% and 100% respectively, and there was no moderate or severe adverse reactions observed. These data showed the vaccines have satisfactory effect on safety and protection.
Animals ; Antibodies, Viral ; blood ; Carrier State ; epidemiology ; veterinary ; Cats ; virology ; Cercopithecus aethiops ; Dogs ; virology ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique ; Rabies Vaccines ; immunology ; Rabies virus ; classification ; genetics ; isolation & purification ; Vero Cells

OBJECTIVETo analyze a Duchenne muscular dystrophy(DMD) patient's muscular regeneration, dystrophin expression and locomotive variation before and after he underwent umbilical cord blood stem cell transplantation in order to assess the therapeutic effect.

METHODSA 12-year-old DMD boy who could not walk for 3 years was confirmed by gene analysis and dystrophin protein immune test on his muscle. He had no other chronic disease. By HLA matching, a piece of umbilical cord blood stem cell with 6 HLA sites matching to the boy was found in Guangdong Umbilical Cord Blood Bank. The number of the nucleated cells of the umbilical cord blood stem cell was 24.08x 10(8). After pretreatment for the DMD boy with busulfan, cyclophosphamide and rabbit anti-human thymocyte globulin, the allergenic cord blood stem cells were transplanted into him by intravenous injection. Cyclosporin A, methylprednisolone, MMF, prostaglandin E1 and ganciclovir were given after the transplantation. At the same time, Gran, the granulocytic cell stimulating factor, and gamma globulin were administered. The biochemistry profile including serum creatine kinase (CK), the reconstruction of blood making, the deletion exon of DMD gene, the regenerating muscles, the dystrophin protein expression, and the locomotive function of the DMD boy were tested regularly.

RESULTS(1) The white blood cells (WBC) of peripheral blood decreased gradually to zero after pretreatment. In a period of 15 days after transplantation, the neutrophil increased to 0.5x 10(9)/L; at 25 days, WBC increased to normal level. Blood platelet was more than 20x 10(9)/L at 22 days. The hemoglobin rose to 85-100 g/L. At 140 days, sternal puncture revealed the rapid growth of neutrophil, blood platelet and hemoglobin. (2)At 140 days, the blood type of the DMD boy transformed from type O to type AB (the donor's blood type being AB). There was no grafe versus host reaction. (3) At 18, 30, 43, 55, 74 and 233 days after transplantation, the PCR-short tandem repeat test of the boy's peripheral blood DNA showed that his genotype was completely the same as the donor's. The results of PCR-short tandem repeat tests of the bone marrow cells DNA by sternal puncture at 140, 183 and 235 days were the same as those of the blood DNA. (4) At 60 days, DMD gene analysis by PCR showed that the defected DMD gene (exon 19 deletion) had been corrected by the umbilical cord stem cells transplantation. (5) At 75 days, the biopsy of calf muscle showed there were myoblast cells and muscular tubes growing. The dystrophin expressions were weak, but a few of them were strong. DNA analysis showed that the donor's gene DNA accounted for 1%-13%. At 126 days, obviously increased dystrophin positive muscular fibers of the boy were found. The donor's fibers rose to 2.5%-25%. (6) The serum CK of the boy declined from 5735 U/L to 274 U/L. (7) At 100 days, physical examination revealed improvement in his arms and legs.

CONCLUSIONThe therapy of Duchenne muscular dystrophy with allogeneic umbilical cord blood hematopoietic stem cell transplantation may reset up the blood-making function, decrease the serum CK level, restore the dystrophin in muscles, and improve the locomotive function of the DMD boy. These data suggest that the allogeneic umbilical cord blood hematopoietic stem cell transplantation may benefit the DMD boys.

Alprostadil ; therapeutic use ; Busulfan ; therapeutic use ; Child ; Combined Modality Therapy ; Cord Blood Stem Cell Transplantation ; methods ; Cyclosporine ; therapeutic use ; Dystrophin ; genetics ; Ganciclovir ; therapeutic use ; Humans ; Male ; Methylprednisolone ; therapeutic use ; Muscular Dystrophy, Duchenne ; genetics ; therapy ; Polymerase Chain Reaction ; Treatment Outcome

Background:From April to July in 2009 and 2010,unexplained severe hemorrhagic fever-like illnesses occurred in farmers from the Huaiyangshan mountains range.Methods:Clinical specimens (blood,urine,feces,and throat swabs) from suspected patients were obtained and stored.Mosquitoes and ticks in affected regions were collected.Virus was isolated from 2 patients and characterized by whole genome sequencing.Virus detection in additional patients and arthropods was done by virus-specific reverse transcription (RT) PCR.Clinical and epidemiological data of RT-PCR confirmed patients were analyzed.Results:An unknown virus was isolated from blood of two patients and from Haemaphysalis ticks collected from dogs.Whole genome sequence analysis identified the virus as a novel member of the family Bunyaviridae,most closely related to the viruses of the genus Phlebovirus within which it forms a separate lineage.Subsequently,infection was confirmed by RT-PCR in 33 of 58 suspected patients.The illness in these patients was characterized by fever,severe malaise,nausea,vomiting,and diarrhea.Prominent laboratory findings included low white cell- and platelet counts,coagulation disturbances,and elevation of liver enzymes.Hemorrhagic complications were observed in 3 cases,5 (15%) patients died.Conclusions:A novel tick-borne Bunyavirus causing life-threatening hemorrhagic fever in humans has emerged in the Huaiyangshan mountain areas of China.Further studies are needed to determine the epidemiology,geographic distribution and vertebrate animal ecology of this virus.

OBJECTIVE: To explore the clinical effect of warming-needle moxibustion with different lengths of moxa stick for asthenospermia with kidney deficiency and liver depression. METHODS: A total of 240 patients with asthenospermia of kidney deficiency and liver depression were randomly divided into a 4-cm group (moxibustion with 4-cm moxa stick, 60 cases, 3 cases dropped off), a 3-cm group (moxibustion with 3-cm moxa stick, 60 cases, 4 cases dropped off), a 2-cm group (moxibustion with 2-cm moxa stick, 60 cases, 2 cases dropped off) and an acupuncture group (60 cases, 3 cases dropped off). All patients were treated with warming-needle moxibustion with different lengths of moxa stick or conventicnal acupuncture at Zhongwan (CV 12), Guanyuan (CV 4), Zhongji (CV 3), Guilai (ST 29), Yaoyangguan (GV 3), Guanyuanshu (BL 26), etc., once a day, five times a week; 4-week treatment was taken as one course, a total of two courses of treatment were given. The semen routine indexes, seminal plasma biochemical indexes, sex hormone levels and TCM syndrome score were compared before and after treatment among the 4 groups. RESULTS: After treatment, the sperm density, sperm viability, ratio of grade A sperm, ratio of grade A and B sperm, seminal plasma fructose and neutral α-glucosidase were higher than those before treatment (P<0.05, P<0.01), and the sperm deformity rates were lower than those before treatment in the 4-cm group and the 3-cm group (P<0.01, P<0.05). The ratio of grade A sperm, ratio of grade A and B sperm, seminal plasma fructose and neutral α-glucosidase in the 4-cm group were higher than the other three groups (P<0.05, P<0.01), and the sperm deformity rate was lower than the other three groups (P<0.05). After treatment, except for dizziness and tinnitus score, each-domain score and total scores of TCM syndrome scale in the 4-cm group and the 3-cm group were lower than those before treatment (P<0.01, P<0.05). The each-domain score of depression, weak waist and knees, low sexual function and total score in the 4-cm group were lower than those in the other three groups (P<0.05, P<0.01). The total effective rate was 87.7% (50/57) in the 4-cm group, which was higher than 78.6% (44/56) in the 3-cm group, 77.6% (45/58) in the 2-cm group and 70.2% (40/57) in the acupuncture group (P<0.05, P<0.01). CONCLUSION The warming-needle moxibustion with 4-cm moxa stick could effectively improve quality and motility of sperm and clinical symptoms in patients with asthenospermia of kidney deficiency and liver depression, which is superior to moxibustion with 3-cm, 2-cm moxa sticks and conventional acupuncture.
Acupuncture Points ; Acupuncture Therapy ; Depression ; Fructose ; Humans ; Kidney ; Liver ; Male ; Moxibustion ; Semen ; Treatment Outcome ; alpha-Glucosidases

Objective: To explore the association between self-reported gingival bleeding and prevalent hypertension among adults in Zhejiang with a cross-sectional study. Methods: After excluding participants with self-reported, physician-diagnosed heart diseases, stroke, diabetes, cancer, and those who never or rarely brush their teeth at baseline study, 48 625 participants aged 30-79 in the China Kadoorie Biobank (CKB) study from Tongxiang, Zhejiang were included for the final analysis. Three multivariable logistic regression models were used to estimate the odds ratios (ORs) for the associations of self-reported gingival bleeding with prevalent hypertension. Results: The mean age of women was (51.2±9.2) years, and 42.9% of participants had prevalent hypertension. The percentage of self-reported frequent gingival bleeding was 6.56% (95%CI: 6.38%-6.75%), significantly higher among women (8.08%, 95%CI: 7.82%-8.35%) than among men (4.36%, 95%CI: 4.12%-4.60%) (P<0.001). After adjusting for socio-demographic factors, behavioral lifestyle, sleep duration, BMI, waist circumference, snoring, in comparison with men whose gingivae never or rarely bleed while brushing teeth, the odds ratio (95%CI) of hypertension for those with occasional, and frequent gingival bleeding were 1.04 (0.96-1.12) and 1.18 (1.02-1.37), respectively (trend P =0.038). The corresponding figures for women were 0.96 (0.91-1.02) and 0.95 (0.86-1.05), respectively (trend P=0.344). Conclusion: Frequent gingival bleeding was positively associated with prevalent hypertension among men.
Adult ; China/epidemiology* ; Cross-Sectional Studies ; Female ; Gingival Hemorrhage ; Humans ; Hypertension/epidemiology* ; Male ; Middle Aged ; Risk Factors ; Self Report