BACKGROUND:Basic fibroblast growth factor (bFGF) plays an important role in the ligament tissue healing process, and bone marrow mesenchymal stem cel s transfected with growth factors can be used as seed cel s in ligament tissue engineering.
OBJECTIVE:To observe biological effect of bone marrow mesenchymal stem cel s transfected with recombinant adenovirus vectors carrying bFGF in three-dimensional co-culture with ligament fibroblasts.
METHODS:Passage 3 bone marrow mesenchymal stem cel s were divided into three groups:control group, Ad-EGFP group and Ad-bFGF group. Under a phase contrast microscope, the changes in cel morphology were observed and the rate of transfection was analyzed by flow cytometry. Proliferation of bone marrow mesenchymal stem cel s and ligament fibroblasts was analyzed by MTS, the expression of bFGF protein in bone marrow mesenchymal stem cel s was determined by ELISA. Scleraxis, col agen type I, col agen type III, decorin and cartilage oligomeric matrix protein levels were detected in BMSCs and ligament fibroblasts using real-time fluorescent quantitative PCR.
RESULTS AND CONCLUSION:Recombinant adenovirus-mediated bFGF gene could transfect bone marrow mesenchymal stem cel s efficiently. After co-culture for 3, 6 days, compared with the control group and Ad-EGFP group, in the Ad-bFGF group, the proliferation ability of bone marrow mesenchymal stem cel s and ligament fibroblasts was enhanced (P<0.01), the expression of bFGF protein in supernatant was obviously higher (P<0.01), the col agen type I, col agen type III, decorin and cartilage oligomeric matrix protein mRNA expression decreased in the ligament fibroblasts (P<0.01), but the mRNA expression of Scleraxis, col agen type I, col agen type III in bone marrow mesenchymal stem cel s increased (P<0.01). The results suggest that the co-culture of Ad-bFGF-transfected bone marrow mesenchymal stem cel s with ligament fibroblasts promotes the proliferation of ligament fibroblasts while decreases the col agen synthesis at the same time, and stimulates the proliferation and differentiation of bone marrow mesenchymal stem cel s into ligament fibrolasts.

Objective To study Kaposi's sarcoma-associated herpesvirus(KSHV)infection in chronic hepatitis B(CHB)patients and its correlation with hepatitis B virus(HBV)replication and treatment-related factors.MethodsEnzyme-linked immunosorbent assay(ELISA)with recombination protein KSHV ORF65 was employed to detect the KSHV antibody and real-time polymerase chain reaction(PCR)was performed to detect KSHV DNA and HBV DNA in CHB patients.Age,HBV replication and licorice preparation treatment of patients were further analyzed.Comparison of rates was done using X~2 test.Results KSHV ORF65 antibody positive rates were 27.3% in 161 male CHB patients and 30.0% in 50 female patients(X~2=0.135,P＞0.05).The KSHV infection rates were increased with age,but this tendency was not obvious in patients older than 40 years old.The highest infection rate was in age group of 31-40 years old which was 37.1%.The positive rate of HBV DNA in CHB patients with KSHV infection was 73.5%,which was 56.3% in uninfected patients(X~2=3.969,P＜0.05).The average plasma level of KSHV DNA in patients treated with licorice preparations was 204.7 copy/mL and that in patients without licorice preparation treatment was 533.9 copy/mL.Eight patients were KSHV DNA positive(KSHV DNA＞ 100 copy/mL)in 16 patients treated with licorice preparations and 23 were positive in 33 patients without licorice preparation treatment.Conclusions The KSHV infection rates are increased with age of CHB patients.KSHV infection may interfere with HBV replication and licorice preparations may suppresss KSHV replication in vivo.

Objective To observe the relation between onset age and the motor complications involved Parkinaon' s disease (PD).Methods The detailed clinical information of 195 patients with idiopathic PD and good response to L-dopa were recorded and followed up.The data were calculated with SPSS statistic software.Results Although the time interval between the onset of the disease and the use of L-dopa was significantly longer in the 57 early-onset patients as compared to the 138 later-onset ones ((3.88±3.33) years vs (2.36±2.21) years, t = 3.142, P = 0.002), the time interval between the use of L-dopa and the occurrence of motor complications was not significantly longer in the early-onset group ((3.81±2.06) years vs (4.24±2.00) years, t = -0.888, P = 0.378).There was no difference in the constituent ratio of non-L-dopa use from onset between early-onset and later-onset PD groups (28.07% (16/57) vs 27.54% (38/138), χ2 = 0.006, P = 0.940).There was also no difference in the dosage of daily L-dopa use when motor complications occurred between early-onset and later-onset PD groups ((601.8± 296.7) mgvs (655.6±192.5) mg, t = -0.912, P=0.365).Seven-tenths of the patients with an onset age younger than 40 years old carried the risk of motor complications after using L-dopa for 5-years and those older than 70 years had the risk at a rate of 1/10.Conclusions Delaying the of use of L-dopa may not necessarily delay the onset of motor complications.The high incidence of motor complications among younger patients may not be related with drug dosage and the type of drug firstly chosen.Younger onset age does inerease the ineidence of motor complicatious.

Objective To compare the validity of four commercially available ELISA kits for detecting HSV-2 type-specific IgG antibodies. Methods A total of 125 serum specimens were collected from 105 patients with genital ulcers and 20 normal individuals without history of STDs. Four ELISA kits which are commercially available in China for the detection of HSV-2 type-specific IgG antibodies were selected for the evaluation. Western blot assay was used as the gold standard. Results Based on the results of detection by Western blot assay, the sensitivity and specificity of these ELISA kits including home-made 1, home-made 2, improted 1 and improted 2 were 13.1% and 98.4%, 7.5% and 100%, 100% and 11.1%, 87.7% and 96.7%, respectively. The areas under the ROC curve of three kits including home-made 1, imported 1 and imported 2 were 0.885 (0.822 - 0.948), 0.852 (0.747 - 0.902), 0.947 (0.950 - 0.998), respectively. Conclusions The results of imported 2 are well consistent with those of Western blot, while the results of other 3 kits are poorly consistent with those of Western blot. It is also indicated that the commercially available ELISA kits for detecting HSV-2 type-specific antibodies should be re-evaluated in terms of their validity befor being applied for the clinical diagnosis as well as laboratory research.

In this study ,we established Cross Priming Amplification (CPA ) technology for detection of influenza A virus (H1N1) approach ,and evaluated the method through clinical specimens .A set of specific primers were designed for CPA ac‐cording to the conservative gene sequences ,designed and realized in the same temperature reverse transcription of RNA and DNA amplification . The amplification products can be totally enclosed nucleic acid detection device for testing . Fourteen healthy pharyngeal swab specimens ,seven other respiratory viruses ,and six arboviruses strains were used as the controls .We used a method that application of gradient dilution to the H 1N1 virus strain as the control to test the sensitivity of the CPA .We also used 102 clinical pharyngeal swab specimens of H1N1 patients for detection object to evaluate the feasibility of CPA clinical detection .Results showed that the CPA reaction did not appear cross reaction on health cases samples and other viruses .The sensitivity of the CPA was approximately 10 copies/uL in the established method that exactly titer H1N1 virus strain gradient dilution test .As to the positive results among the clinical pharyngeal swab samples collected from patients at different stages after onset ,the CPA had the highest positive detection rate during the first three days after onset (100% ) .While the detection rate from day 4 to day 6 after onset was 79 .31% .After 7 days ,the detection rate was 9 .09% .The established CPA assay was a highly sensitive ,specific and reproducible approach for rapid detection of H1N1 virus ,which is conducive to the early diagno‐sis of influenza A virus (H1N1) for basic medical units .

A bacterium, having high chitinolytic activity, was isolated from sediment of Shantou Bay, named SWCH-6. According to its physiological and biochemical characteristics and 16S rDNA sequence, it was identified as Aeromonas hydrophlilla. The optimal chitinase fermentation conditions of strain SWCH-6 were conformed by single-factor experiments and orthogonal experiments, they were colloidal chitin 25.0 g/L, tryptone 10.0 g/L, seawater 1 L, pH 8.5, 32 ℃ , 150 r/min for 72 h. In these conditions, its chitinase activity reached 0.39 U/mL. In addition, at 40℃ and pH 5.0, its chitinase performed the highest catalytic activity and its chitinase activity could be enhanced by Cu2+, Fe3+ and surfactant toween-80; weakened by Zn2+, Mn2+ and surfactants SDS, detergent powder, and there were some differences from orther chitinases.

Objective To explore the influence of delivery mode and feeding pattern on cytomegalovirus (CMV) infection on infants born ≥ 32 gestational weeks,and to observe the outcomes after CMV infection.Methods In this retrospective study,378 pregnant women with positive CMV IgG and negative CMV IgM,and their offsprings (384 cases,including six pairs of twins),who got visited at five hospitals of our collaboration group during March 2013 and February 2016,were enrolled.Serum samples were retrieved from a previous study of these participants for CMV IgM and IgG detection with enzyme-linked immunosorbent assay.All participants were divided into exclusive artificial feeding (EAF) and breastfeeding groups (BF),and the latter included exclusive breastfeeding (EBF) and mixed feeding (MF).T or Chi-square or Fisher's exact tests were performed for statistical analysis.Results (1) Among the 378 pregnant women,there were 186 mothers and 190 infants (4 pairs of twins) in BF group,and the other 192 mothers and 194 infants (2 pairs of twins) in EAF group.The percentage of male infants were 54.7％(104/186) and 56.2％(109/194) in the BF and EAF group,respectively.The mean birth age was (38.9± 1.4) and (38.7± 1.7) weeks,and the age at followingup was (9.8± 2.2) and (10.5± 2.9) months,respectively.(2) The CMV IgG positive rate of infants in BF group was higher than in the EAF group [62.6％(119/190) vs 29.9％ (58/194),x2=41.403,P＜0.001].CMV IgG levels in infants were higher than the mothers [(537.1 ±249.5) vs (416.2±241.2) U/ml,t=4.609,P＜0.001].In infants with positive CMV IgG,the positive rates of CMV IgM were similar in the two groups [21.0％(25/119) vs 19.0％ (11/58),x2=0.101,P=0.751].(3) The positive rate of CMV IgG in vaginally born infants was higher than those born by caesarean section [55.2 (95/172) vs 38.7％ (82/212),x2=10.472,P=0.001].Further analysis in the EAF group showed that those infants born vaginally had a higher positive rate ofCMV IgG than those born by caesarean section [42.9％ (33/77) vs 21.4％ (25/117),~=10.231,P=0.001],while this figure did not show statistical difference in the BF group.(4) Infants with positive or negative CMV IgG were in similar age and gender proportion,as well as their height and weight.Among 36 infants with both positive CMV IgG and IgM,three failed in alanine aminotransferase (ALT) test due to hemolysis.However,among the other 33 cases,15.1％ (five cases) presented with lightly elevated ALT (42-107.2 U/L),which was similar to those infants with positive CMV IgG and negative CMV IgM (14/98,14.3％) and those with both negative CMV IgG and IgM (20/144,13.9％),(x2=0.036,P=0.982).Conclusions Although breastfeeding and vaginal birth may increase CMV infection rate in neonates and infants,but no obviously adverse prognosis was reported in those born over 32 gestational weeks.So we should encourage vaginal birth and breastfeeding in these population.

Based on the Chinese pharmacopeia 2000 ed and Japanese pharmacopeia 14st ed., the original plant differencesof Chinese raditional medicines and Japanese Kampo medicines were compared by making list. The differences and reasons were analyzed.
China ; Drugs, Chinese Herbal ; Japan ; Medicine, Chinese Traditional ; Medicine, Kampo ; Pharmacopoeias as Topic ; Plants, Medicinal

Objective To evaluate the immune effects of bivalent inactivated rotavirus vaccine (IRV) and investigate the viability of development of bivalent IRV.Methods Firstly,bivalent IRV was prepared by mixing G1 IRV and G3 IRV with equal amount,G1 IRV and G3 IRV as monovalent control,PBS as negative control.Secondly,those vaccines were vaccinated to the mice by intramuscular injection.Then,to evaluate the immune effects of bivalent IRV,the levels of serum or fecal rotavirus specific IgG and IgA were assessed by ELISA,the levels of serum neutralized antibody were measured by microneutralization assay,the number of IFN-γ or IL-4 secreting cells were analyzed by ELISPOT assay.Results Compared to negative control group,bivalent IRV induced the higher levels of serum and fecal G1 and G3 rotavirus specific antibody.It was found that there were no significant differences for the levels of serum IgG and IgA,fecal IgG and IgA,serum neutralized antibody between induced by bivalent IRV and induced by G1 type monovalent vaccines ; but there were significantly increase for the levels of serum IgG (t =2.691,P＜0.05) and serum neutralized antibody (t =2.561,P＜0.05) between induced by bivalent IRV and induced by G3 monovalent vaccines,there were no significant differences for other antibodies between induced by bivalent IRV and induced by G3 monovalent vaccines.At the same time,compared to negative control group,bivalent IRV induced significantly increase in the number of IFN-γ or IL-4 secreting cells in spleen lymphocytes.It was found that there were no significant differences for the number of IFN-γ or IL-4 secreting cells stimulated by G1 rotavirus between bivalent IRV and G1 monovalent vaccines; but there were significantly increase for the number of IL-4 secreting cells (t =2.327,P＜0.05) stimulated by G3 rotavirus between bivalent IRV and G3 monovalent vaccines,there were no significant differences for the number of IFN-γ secreting cells stimulated by G3 rotavirus between bivalent IRV and G3 type monovalent vaccines.Conclusion The bivalent IRV can induce effective immune response,in which there were no inhibitory interference between the components of bivalent IRV,which provided the experimental basis for the development of bivalent IRV.

This study aimed to simultaneously determine the contents of astilbin and engeletin in dong medicine "sunl gaems" of Guizhou origin by quantitative analysis of multi-components by single marker (QAMS), with astilbin as the internal standard substance. On UPLC and HPLC chromatograms, different models of instruments were used to investigate relative correction factors (RCF), in order to discuss the interoperability of RCFs established in different chromatographic systems. The engeletin content was calculated based on the established RCFs and compared by the one point external standard method and the external standard working curve method, in order to verify the accuracy of QAMS. According to the result, in different chromatograms, the ratios between RCF and retention time of engeletin and astilbin had a good reproducibility, with RSD between 2.0% and 1.8%, both were less than 3%. The relative differences among results of QAMS, the external standard working curve method of dong medicine "sunl gaems" ranged between 1.6% and 3.9%, with RSD between 2.02%-0.80% in line with relevant requirements and Pearson correlation coefficient at 0.9998 (P <0.01). The findings showed that QAMS was an accurate, reliable and highly reproducible method to determine the contents of astilbin and engeletin in dong medicine "sunl gaems" of Guizhou origin and so could be used to control the inherent quality of the herb.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Flavonols ; analysis ; Glycosides ; analysis