ObjectiveTo investigate the effects of exogenous Fusarium oxysporum and Trichoderma viride on the diversity of soil fungal community and carbon metabolic function of cultivated Panax ginseng. MethodIllumina HiSeq 2500 high-throughput sequencing combined with Biolog-ECO was used to analyze the species diversity and functional diversity of soil fungal communities in P. ginseng soil under different exogenous treatments. ResultThe results of high-throughput sequencing showed that the number and species of microorganisms in the soil were significantly changed after exogenous microorganisms were added. The soil fungi with relative abundance greater than 1% included Mortierella sp.，Fusarium sp.，Humicola sp.，and Simplicillium sp. Mortierella sp. in each treatment group significantly increased. Humicola sp. and Simplicillium sp. could be induced to increase by exogenous addition of F. oxysporum，while T. viride at a high concentration could significantly inhibit the growth of F. oxysporum. As revealed by Biolog and principal component analysis （PCA），the average well color development （AWCD） in the high-dose T. viride group （MG） was significantly higher than that in the control group （QS）and the low-dose F. oxysporum group（LD）. The utilization abilities for amino acids，carboxylic acids，polymers， and amines were enhanced in the MG group，but the microbial metabolic activity was reduced in the high-dose F. oxysporum group （LG）. There was no significant increase in the utilization of phenolic acids by soil microorganisms in both groups. ConclusionExogenous addition of F. oxysporum can lead to the growth and reproduction of other pathogenic fungi. Exogenous addition of T. viride can enhance the soil fungal community structure and metabolic diversity，inhibit the proliferation of F. oxysporum，and improve the soil microbial environment of cultivated P. ginseng.

This study aimed to investigate the effect and the possible mechanism of Shenlian( SL) extract on tumor necrosis factor-α( TNF-α)-induced ECV304 injury. After the establishment of TNF-α-induced ECV304 cells injure model,MTT assay was used to detect cell viability and the level of reactive oxygen species( ROS) was measured by flow cytometry. The contents of superoxide dismutase( SOD),malondialdehyde( MDA),nitric oxide( NO),endothelin-1( ET-1) and interleukin-1β( IL-1β) in the supernatant were detected by biochemical method and enzyme linked immunosorbent assay( ELISA). The expression levels of apoptosis-related proteins B-lymphoma-2 gene( Bcl-2),Bcl-2 associated X protein( Bax),caspase-3,caspase-9 and nuclear factor E2 associated factor2( Nrf2)/Kelch like epichlorohydrin associated protein-1( Keap1) signaling pathway related proteins Nrf2,Keap1,quinone oxidoreductase( NQO1) and heme oxygenase 1( HO-1) were detected by Western blot. The results showed that 50 μg·L-1 TNF-α significantly damaged ECV304 cells,induced the impairment of cell viability( P<0. 01),the increase of ROS production,the decrease of SOD activity,and the increase of MDA,NO,ET-1 and IL-1β( P<0. 01),meanwhile,it caused the up-regulation of Keap1,caspase-9 and Bax protein expression,and down-regulation of NQO1 and Bcl-2 protein expression( P<0. 05) compared with the control group.Compared with the model group,SL extract reduced the damage of ECV304 cells induced by TNF-α,improved cell viability,reduced ROS production,increased SOD activity and decreased MDA,NO,ET-1,IL-1β content( P<0. 01 or P<0. 05). In addition,SL extract also down-regulated the protein expression levels of Keap1,caspase-3,caspase-9 and Bax,and increased the protein expressions of Nrf2,NQO1,HO-1 and Bcl-2( P<0. 01 or P<0. 05). The above results indicate that SL extract can provide protective effect on ECV304 cells injury induced by TNF-α,alleviate oxidative stress injury,inflammation and apoptosis,and its mechanism may be related to regulating Nrf2/Keap1 signaling pathway.
Apoptosis ; Kelch-Like ECH-Associated Protein 1/metabolism* ; NF-E2-Related Factor 2/metabolism* ; Oxidative Stress ; Plant Extracts ; Signal Transduction ; Tumor Necrosis Factor-alpha/genetics*

OBJECTIVE: To investigate the risk factors for cow's milk protein allergy (CMPA) among infants through a multicenter clinical study. METHODS: A total of 1 829 infants, aged 1-12 months, who attended the outpatient service of the pediatric department in six hospitals in Shenzhen, China from June 2016 to May 2017 were enrolled as subjects. A questionnaire survey was performed to screen out suspected cases of CMPA. Food avoidance and oral food challenge tests were used to make a confirmed diagnosis of CMPA CMPA. A multivariate logistic regression analysis was used to investigate the risk factors for CMPA. RESULTS: Among the 1 829 infants, 82 (4.48%) were diagnosed with CMPA. The multivariate logistic regression analysis showed that maternal food allergy (OR=4.91, 95%CI: 2.24-10.76, P<0.05), antibiotic exposure during pregnancy (OR=3.18, 95%CI: 1.32-7.65, P<0.05), and the introduction of complementary food at an age of <4 months (OR=3.55, 95%CI: 1.52-8.27, P<0.05) were risk factors for CMPA, while exclusive breastfeeding (OR=0.21, 95%CI: 0.08-0.58, P<0.05) and the introduction of complementary food at an age of >6 months (OR=0.38, 95%CI: 0.17-0.86, P<0.05) were protective factors. CONCLUSIONS The introduction of complementary food at an age of <4 months, maternal food allergy, and antibiotic exposure during pregnancy are risk factors for CMPA in infants.
Animals ; Cattle ; China ; Female ; Humans ; Infant ; Milk Hypersensitivity ; Milk Proteins ; Pregnancy ; Risk Factors ; Surveys and Questionnaires

Objective To compare the values of dynamic enhanced magnetic resonance imaging(DCE-MRI),digital breast tomosynthesis(DBT),and digital mammography(DM)in the early detection and diagnosis of breast cancer.Methods We retrospectively analyzed the clinical and imaging data of 65 cases with early breast cancer confirmed by surgical pathology from June 2017 to December 2018.All patients underwent breast DCE-MRI,DM and DBT before surgery.The receiver operating characteristic(ROC)curves were drawn,with the pathological results as the gold standard,to evaluate the diagnostic performance of different examination methods.The areas under ROC curves(AUCs)were compared using test.The differences among DCE-MRI,DBT and DM in detecting early breast cancer were compared using chi-square test in terms of positive rates,accuracy,sensitivity,and specificity.Pearson correlation analysis was performed to assess the accuracy of these imaging methods in detecting the size of early breast cancer.Results The AUCs of DCE-MRI,DBT,and DM based on the BI-RADS classification for early diagnosis of breast cancer were 0.910,0.832,and 0.700,respectively(=2.132,=0.001);the sensitivity of DCE-MRI,DBT,and DM for early breast cancer was 92.3%,70.8%,and 52.5%,the specificity was 65.0%,85.0%,and 79.3%,and the accuracy was 83.1%,70.8%,and 50.8%,indicating that DCE-MRI(=15.330,=0.0001) and DBT(=5.450,=0.020) had significantly higher diagnostic accuracy than DM.The measurement results of DM,DBT,and DCE-MRI were positively correlated with the pathological measurements(=0.781,=0.847,=0.946;all <0.01). Conclusions DCE-MRI and DBT have higher positive rates and accuracies than DM in detecting early breast cancer.Medical institutions where DCE-MRI is still not available can use DBT to improve the early detection of breast cancer.
Breast ; diagnostic imaging ; Breast Neoplasms ; diagnostic imaging ; Female ; Humans ; Magnetic Resonance Imaging ; Mammography ; methods ; Retrospective Studies

Objective To investigate the sub-cellular distribution correlation between activated LIMK1 (pLIMK1Thr508) and Aurora-A in mouse oocyte meiosis,and changes in Aurora-A location and spindle structure in condition of LIMK1 inhibition.Methods Immunofluorescence staining was employed to detect the sub-cellular localization of pLIMK1Thr508and its spatial-temporal correlation with spindle organizing regulator Aurora-A in mouse oocyte meiosis; BMS-3, the specific inhibitor to LIMK1 activity, was applied to analyze the effects of LIMK1 inhibition on Aurora-A distribution and spindle formation. Results At meiotic prophase,pLIMK1Thr508was weakly detected and concentrated in the germinal vesicle(GV) in oocytes,with no signal of Aurora-A across the cytoplasm and nuclear area;as meiotic assumption approaching,pLIMK1Thr508left nuclear,aggregating as a single dense dote in the vicinity of nuclear, and being co-localized with the emerging Aurora-A; After germinal vesicle broke down (GVBD), pLIMK1Thr508and Aurora-A remained overlapped and concentrated as multi foci around the condensed chromosomes;at metaphase Ⅰ(MⅠ) and metaphase Ⅱ(MⅡ), pLIMK1Thr508was co-localized with Aurora-A on spindle poles;During anaphase Ⅰ(AⅠ) to telophase Ⅰ(Tel Ⅰ) progression, pLIMK1Thr508was detached from spindle poles and mainly concentrated on the cleavage furrow,while Aurora-A loosely congressed on spindle. In ad-dition, LIMK1 inhibition with BMS- 3 destroyed Aurora-A polar location and spindle formation. Conclusions pLIMK1Thr508is a microtubule organizing center (MTOC)-associated protein, may participate in spindle assembly and maintenance through regulating Aurora-A in mouse oocytes during meiotic progression.

BACKGROUNDKnee osteoarthritis (KOA) is a chronic joint disease that manifests as knee pain as well as different degrees of lower limb swelling, stiffness, and movement disorders. The therapeutic goal is to alleviate or eliminate pain, correct deformities, improve or restore joint functions, and improve the quality of life. This study aimed to evaluate the efficacy and safety of Zhuanggu joint capsules combined with celecoxib and the benefit of treatment with Zhuanggu alone for KOA.

METHODSThis multi-center, randomized, double-blind, double-dummy, parallel controlled trial, started from December 2011 to May 2014, was carried out in 6 cities, including Beijing, Shanghai, Chongqing, Changchun, Chengdu, and Nanjing. A total of 432 patients with KOA were divided into three groups (144 cases in each group). The groups were treated, respectively, with Zhuanggu joint capsules combined with celecoxib capsule simulants, Zhuanggu joint capsules combined with celecoxib capsules, and celecoxib capsules combined with Zhuanggu joint capsule simulants for 4 weeks consecutively. The improvement of Western Ontario and McMaster Universities Osteoarthritis (WOMAC) index and the decreased rates in each dimension of WOMAC were evaluated before and after the treatment. Intergroup and intragroup comparisons of quantitative indices were performed. Statistically significant differences were evaluated with pairwise comparisons using Chi-square test (or Fisher's exact test) and an inspection level of α = 0.0167.

RESULTSFour weeks after treatment, the total efficacies of Zhuanggu group, combination group, and celecoxib group were 65%, 80%, and 64%, respectively, with statistically significant differences among the three groups (P = 0.005). Intergroup pairwise comparisons showed that the total efficacy of the combination group was significantly higher than that of the Zhuanggu (P = 0.005) and celecoxib (P = 0.003) groups. The difference between the latter two groups was not statistically significant (P > 0.0167). Four weeks after discontinuation, the efficacies of the three groups were 78%, 95%, and 65%, respectively, with statistically significant differences (P < 0.0001). Intergroup pairwise comparisons revealed that the efficacy of the combination group was significantly better than that of the Zhuanggu and the celecoxib groups (P < 0.0001). The difference between the latter two groups was not statistically significant (P > 0.0167). The incidences of adverse events in Zhuanggu group, combination group, and celecoxib group were 8.5%, 8.5%, and 11.1%, respectively, with insignificant differences (P > 0.05).

CONCLUSIONSZhuanggu joint capsules alone or combined with celecoxib showed clinical efficacy in the treatment of KOA. The safety of Zhuanggu joint capsules alone or combined with celecoxib was acceptable.

TRIAL REGISTRATIONChinese Clinical Trial Registry, ChiCTR-IPR-15007267; http://www.medresman.org/uc/project/projectedit.aspx?proj=1364.

Adult ; Aged ; Celecoxib ; administration & dosage ; adverse effects ; Double-Blind Method ; Drug Therapy, Combination ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; drug therapy

In vitro amplified human leukocyte antigen (HLA)-haploidentical donor immune cell infusion (HDICI) is not commonly used in children. Therefore, our study sought to evaluate its safety for treating childhood malignancies. Between September 2011 and September 2012, 12 patients with childhood malignancies underwent HDICI in Sun Yat-sen University Cancer Center. The median patient age was 5.1 years (range, 1.7-8.4 years). Of the 12 patients, 9 had high-risk neuroblastoma (NB) [7 showed complete response (CR), 1 showed partial response (PR), and 1 had progressive disease (PD) after multi-modal therapies], and 3 had Epstein-Barr virus (EBV)-positive lymphoproliferative disease (EBV-LPD). The 12 patients underwent a total of 92 HDICIs at a mean dose of 1.6×10(8) immune cells/kg body weight: 71 infusions with natural killer (NK) cells, 8 with cytokine-induced killer (CIK) cells, and 13 with cascade primed immune cells (CAPRIs); 83 infusions with immune cells from the mothers, whereas 9 with cells from the fathers. Twenty cases (21.7%) of fever, including 6 cases (6.5%) accompanied with chills and 1 (1.1%) with febrile convulsion, occurred during infusions and were alleviated after symptomatic treatments. Five cases (5.4%) of mild emotion changes were reported. No other adverse events occurred during and after the completion of HDIDIs. Neither acute nor chronic graft versus host disease (GVHD) was observed following HDICIs. After a median of 5.0 months (range, 1.0-11.5 months) of follow-up, the 2 NB patients with PR and PD developed PD during HDICIs. Of the other 7 NB patients in CR, 2 relapsed in the sixth month of HDICIs, and 5 maintained CR with disease-free survival (DFS) ranging from 4.5 to 11.5 months (median, 7.2 months). One EBV-LPD patient achieved PR, whereas 2 had stable disease (SD). Our results show that HDICI is a safe immunotherapy for childhood malignancies, thus warranting further studies.
Child ; Child, Preschool ; Cytokine-Induced Killer Cells ; immunology ; Epstein-Barr Virus Infections ; therapy ; Female ; Follow-Up Studies ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Immunotherapy, Adoptive ; Infant ; Killer Cells, Natural ; immunology ; Lymphoproliferative Disorders ; therapy ; virology ; Male ; Neuroblastoma ; therapy ; Transplantation, Homologous ; Treatment Outcome

In order to analyze the molecular epidemiology of Hantavirus (HV) in Zhejiang Province, the complete M and S genome sequences of 12 HV strains from different hosts and locations in Zhejiang Province of China during the period of 1981-2007 were analyzed on genetic evolution by DNAstar and MEGA 4.0 software in this research. Phylogenetic analyses revealed that HTN and SEO strains were co-circulating in Zhejiang Province, and the difference in sequence similarity and the phylogeny was closely related to the isolated regions, but had no distinct relationship with the isolate year and the host, indicating a relationship between epidemiology of HFRS and the distribution region, especially in HTNV. The isolates in the same region could be assigned in same or near phylogenetic clade sharing high sequence similarity. Interestingly, the Gou3 strain and ZJ5 strain isolated from Jiande region in Zhejiang Province formed a distinct phylogenetic lineage in SEOV clade, and different from the other SEOV variants outside China. We believed that the special SEOV variants were distributed in Jiande region.
Animals ; China ; Disease Reservoirs ; virology ; Evolution, Molecular ; Hantavirus ; classification ; genetics ; isolation & purification ; Hantavirus Infections ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Rodentia ; virology ; Viral Proteins ; genetics

Objective To investigate the status of infection and distribution of rabies virus (RV) in different epidemic areas in China. Methods Brain specimens from animals and suspected patients were collected at the districts of high-, medium- and low incidence rates of human rabies and detected by both direct Immunofluorescence assay (DFA) and RT-PCR. Results 254 of 3007 specimens of dog brains showed RV positive by DFA (positive rate of 8.4% ). Among these 254 samples, 78 showed positive (positive rate of 30.7% ) by RT-PCR. 93 specimens from dogs and cats that had attacked human beings, 63 of them showed positive by DFA (positive rate of 67.7%) and all of them were also positive by RT-PCR. In addition, RV could also be detected in Apodemus agrarius,ferret badger, and suspected patients specimens from the districts under survey. There was no statistical difference between the infection rates of RV in different provinces and regions with different incidence of rabies. Conclusion There might be a relatively high infection rate of RV among the domestic dogs/cats in the endemic areas in China. Wild animals might have been infected with RV in the districts under survey.

Objective To establish a TaqMan based real-time reverse transeription-polymerase chain reaction (RT-PCR) assay for the detection of Japanese encephalitis virus. Methods The gene sequences of Japanese encephalitis virus downloaded from the GenBank was aligned, using the biologic software. Specific primers and probes were designed in the conserved region of the C gene for Japanese encephalitis virus. The real-time RT-PCR reactive condition was optimized and the sensitivity, specificity and the stability of the assay were evaluated. Mosquitoes collected from Zhejiang province were detected by this assay. Results Mg2+, primer and probe were optimized at 5 mmol/L, 0.2 μmol/L and 0.1 μmol/L respectively. The specificity of the assay was high and there were no cross reactions with dengue virus, rabies virus, seoul virus or hantan virus. The detection limits of the assay was 0.1 TCID50. Results from preliminary application showed that TaqMan RT-PCR for Japanese encephalitis virus was sensitive, easier and faster to perform the process of traditional virus isolation and identification. It took only three hours to extract viral RNA and perform the real-time RT-PCR. Conclusion This TaqMan-based one-step RT-PCR assay was a quick, sensitive and specific tool for molecular diagnosis of Japanese encephalitis virus.