Objective:To investigate the effect of Bitongling Granule medicated serum on rabbit articular chondrocytes cultured in vitro.Methods:Bitongling Granule(2ml/kg) was given to New Zealand rabbits for 3 days for preparation of containing serum.Rabbit.chondrocytes were divied into Bitongling Granule groups of different concentrations,blank serum group,experimental control group and anti-bone capsule group.To observe the change of nitric oxide synthase and superoxide dismutase in all groups.Results:Bitongling Granule can reduce the activity of NO and increase the level of SOD in sera.Conclusions:Bitongling Granule can prevent and cure experimental osteoarthritis,which is related to the decrease of NO and activation of SOD.

Objective To study the mechanisms of Re Bi granule on treating RA.Methods Make SD maleness rats Adjuvant-Induced Arthritis(AA)which was derived by Freund's complete adjuvant(CFA)as investigation object.Make Wang Bi granule as positive control medicine,and divide it into six groups(blank group,model group,control group,Infusion of Re Bi granule high dose group and midst dose group and low dose group).Observe Re Bi granule's influence and mechanism to synovial cell apoptosis and T cell subsets.Results Re Bi granule could distinctly heighten synovial cell apoptosis ratio of AA rats,heighten CD8+cell's number and reduce CD4+cell's number,lower CD4+/CD8+ ratio,adjust T cells for two-way.Conclusion Re Bi granule not only is an effective treatment way for rheumatoid arthritis,but also can improve the abnormal immune status.

Objective: To study the clinical effects of acupuncture therapy for acute stroke. Methods: 150 patients suffered from acute stroke were randomly divided into two groups: drug group, acupuncture group. To apply routine drug treatment of Neurology for drug group: pisacetam 250mL, Salvia injection 20mL added into normal sodium 250mL for intravenous drip, once a day. For acupuncture group applied calm the liver and suppress yang acupuncture therapy beside drugs. Modified Scandinavian Stroke Scale (SSS) and activities of daily living (ADL) scale (Barthel Index) were used for evaluation, as well as biochemistry\hemorheology determination. Results: The scale showed significant difference as compared with the drug group, biochemistry index had non-significant difference, and hemorheological index had partly improved. Conclusion: The scale could synchronously reflect the clinical symptom and changes of patient with stroke.Acupuncture therapy has a significant improvement of acute stroke treatment.

Objective To investigate the clinical features of internal carotid steal syndrome. Methods The clinical manifestations, CT or MRI, digital subtraction angiography, and blood flow compensation in 6 patients with internal carotid steal syndrome were analyzed. Results Of the 6 patients, 2 had unilateral internal carotid artery stenosis, 4 had severe stenosis (in which 2 were on the left side, 1 was on the right side, and 1 was on both sides). The clinical manifestations of the patients with internal carotid steal syndrome were watershed infarction and transient ischemic attack. Four patients had posterior circulation ischemia and 2 had anterior circulation ischemia. Digital subtraction angiography demonstrated that collateral circulation was established in all the 6 patients. The anterior communicating artery, posterior communicating artery, and pial artery were the common compensatory vessels. Conclusions Internal carotid artery steal syndrome can be presented as anterior or posterior circulation ischemia, and the collateral circulation plays an important role in the compensation.

Objective To analyze the genomic sequences of Cryptococcus neoformans var grubii strains of two genotypes with different virulence and to screen out the virulence-associated genes. Methods A clinical strain (IFM56800) with the strongest virulence and an environmental strain (IFM56731) with the weakest virulence were screened out for whole genome sequencing analysis. The results of sequencing analy-sis were comprehensively analyzed by using the method of comparative genomics. Genetic variations were ex-tensively screened by using the strategies of non-synonymous single nucleotide polymorphisms ( nsSNPs), nonsense SNPs and the insertions or deletions ( InDels) causing frameshift mutations. The filtered genes were sequenced in 20 experimental strains. The whole RNAs were extracted and then the full-length cDNAs were sequenced by using the rapid amplification of 5′ and 3′ cDNA ends (RACE) method. Results By whole genome sequencing, valid data with high coverage (127 times and 111 times) was obtained in both the environmental strain IFM56731 and the clinical strain IFM56800. The data of InDels and SNPs were statisti-cally analyzed, respectively. Six genes were chosen for further analysis based on the strategies of nonsense SNPs and the InDels causing frameshift mutations. The six genes were amplified and sequenced in all of the experimental strains, three of which were further analyzed with cDNA sequencing. Ultimately, the location and structure of CNAG_01032 gene were determined. The predicted nonsense mutation locus was verified to present in the actual mRNA. Conclusion The strategies of nonsense SNPs and the InDels causing frame-shift mutations showed high-efficiency in screening potential virulence-associated genes. The CNAG_01032 gene was screened out as a novel virulence-associated gene.

Objective: To observe the effect of acupuncture on the intestinal flora in Parkinson disease (PD) model mice and explore the mechanism of acupuncture in improving the locomotor function in PD. Methods: Thirty-two C57BL/6 mice were randomly divided into a control group, a 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine (MPTP) group, a MPTP + acupuncture group (MPTP+A), and a MPTP + madopar group (MPTP+M), with 8 mice in each group. Except for the control group, the other groups were intraperitoneally injected [25 mg/(kg·bw)] with MPTP to establish PD mouse models. After successful modeling, the MPTP group received no intervention, the MPTP+A received acupuncture at Tianshu (ST25), Guanyuan (CV4), and Zusanli (ST36), and the MPTP+M was given madopar [125 mg/(kg·bw)] by intragastric gavage. After consecutive 10-day interventions, the intestinal function and behaviors of the mice were detected. The 16S rRNA gene sequence was used to analyze the composition of fecal intestinal flora in each group. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay were used to detect the expression levels of inflammatory cytokines in the brain and serum. The expression levels of tyrosine hydroxylase (TH) and α-synuclein in the substantia nigra (SN) were detected by immunohistochemical staining. Toll-like receptor (TLR) 2 and lipopolysaccharide receptor CD14 (CD14) in the SN were determined by RT-qPCR. Myeloid differentiation factor (MyD) 88, nuclear factor kappa-B (NF-κB) and Akt1 in the SN were detected by Western blotting. Results: After the intervention, compared with the control group, the intestinal motility, fecal water content, and the expression of TH in the SN were significantly decreased in the MPTP group (P<0.05), along with an increased α-synuclein expression (P<0.05). Additionally, the results of the fecal microflora test showed that the alpha diversity of the MPTP decreased, and the levels of inflammatory cytokines [tumor necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS), interleukin (IL)-1β, and IL-6] in the serum and SN, and the expression of NF-κB in the SN were significantly increased (P<0.05). Compared with the MPTP group, acupuncture intervention significantly enhanced the autonomous horizontal movement and coordination ability of PD mice (P<0.05); acupuncture and madopar interventions significantly reduced the levels of α-synuclein, inflammatory cytokines (TNF-α, iNOS, IL-1β, and IL-6) in the serum and SN, and the NF-κB expression in the SN, along with significantly increased alpha diversity richness index (P<0.05). In addition, the relative abundance of Bacteroides increased significantly in the MPTP+A (P<0.05), while the relative abundance of Firmicutes and Cyanobacteria decreased significantly (P<0.05). Conclusion: Acupuncture intervention can improve locomotor function, reduce α-synuclein aggregation and inflammatory factors expression, and increase the Akt signaling pathway in PD mice. In addition, acupuncture intervention can benignly regulate the intestinal flora of PD mice. Therefore, it suggests that acupuncture intervention can protect PD model mice probably by regulating intestinal flora and activating Akt signaling pathway.

Objective To investigate the genetic relation between Cryptococcus neoformans var.the clinical strains in MLMT - 13 genotype and the environmental strains in MLMT - 36 genotype. Methods Multilocus microsatellite typing (MLMT) method was applied for the genotype analysis in our study.Through this method, we recognized two genotypes that distinguish a majority of clinical and environmental strains. In order to compare virulence between the two types, we chose to infect BALB/c mice (6 weeks,female) with 9 MLMT-13 strains and 10 MLMT-36 strains intravenously. Results Forty( 17 clinical and 23 environmental isolates) were analyzed. Of 17 clinical strains, 9 belonged to a major type of MLMT-13 (52.9%). They were mainly isolated from clinical specimens. About 43.5% of strains from the environment belong to a major type of MLMT-36, which are indigenous to environments and which were not isolated from clinical samples. The mortality rate and pathological changes of the above mice were observed during two months after injection. The results showed that the mortality rate of mice infected with MLMT-13 strains was 100%, while the mortality rate with MLMT-36 strains was 7. 5%. The pathological sections showed that lesions of MLMT-13 infected mice appeared in the brain, lungs, liver and kidneys, while the lesions of MLMT-36 infected mice only appeared in the brain. Most brains of MLMT-13 infected mice were distorted,and both the number and size of lesions in such brains were much larger than those of MLMT-36 infected mice. Conclusion Our study illustrated the virulent difference between MLMT-13 and MLMT-36, which are isolated from patients and environment respectively. The results inferred that some genetic changes, such ss microsatellite repeats, might occur between environmental and clinical isolates through their environmental adaptation progress.

Objective To observe the correlation between degrees ofextracranial cerebral artery stenosis and pressure ratio across stenosis.Methods The patients with ≥ 50％ stenosis,admitted to our hospital and conformed by DSA from September 2015 to February 2017,were collected in this study.Collateral circulation compensation was assessed by DSA.The distal stenosis pressure (Pd) and stenosis pressure (Pa) were measured by pressure wire system;the pressure ratio across stenosis was recorded.Correlations of pressure ratio with stenosis rate and collateral circulation compensation were analyzed by Spearman correlation.Results Thirty one stenotic extracranial cerebral arteries (9 of carotid stenosis,12 of vertebral artery stenosis and 10 of subclavian artery stenosis) in 28 patients were collected.There were no perioperative complications or adverse events.The vascular stenosis rate showed negative correlation with pressure ratio (r=-0.615,P=0.000).In the subgroups,pressure gradient was negatively correlated with carotid artery stenosis rate (r=-0.948,P=-0.000),vertebral artery stenosis rate (r=-0.757,P=0.004) and subclavian artery stenosis (r=-0.759,P=0.011).Pressure ratio and collateral circulation compensation showed negative correlated relation (r=-0.475,P=0.007).Conclusion The pressure ratio across stenosis gets worse impairment with increase of severity of stenosis,and the collateral circulation influences the pressure ratio.

Objective To investigate the prevalence of mcr-1 gene,a plasmid-mediated polymyxin resistance gene,in Escherichia coli(E.coli) strains isolated in Dongyang of Zhejiang Province and to under-stand the epidemiological characteristics of E.coli strains carrying mcr-1 gene in order to provide local clini-cians with a theoretical basis for prevention and control of the spread of mcr-1-bearing E.coli strains. Meth-ods A total of 315 E.coli strains were collected in the People′s Hospital of Dongyang, Zhejiang Province from January to December 2016. All strains were isolated from specimens of blood,urine,respiratory tract, etc. PCR was performed to detect the genes confering resistance to polymyxin (mcr-1 gene), β-lactamase and carbapenem. Minimal inhibitory concentrations (MIC) of antibiotics against mcr-1-positive strains were determined by micro-broth dilution method. Conjugation test was performed to confirm whether the mcr-1 gene was located on the transferable plasmid. Multilocus sequence typing (MLST) was used for molecular typing of mcr-1-positive strains. Results Five mcr-1-positive strains were identified from 315 E.coli strains with a positive rate of 1.6%. Two out of the five mcr-1-positive E.coli strains contained β-lactamase resist-ance genes,blaTEM-1and blaCTX-M-14. Both of them were resistant to the first, second and third generation of cephalosporins and one was also resistant to cefepime. All of the five mcr-1-positive E.coli strains were sen-sitive to ciprofloxacin and levofloxacin,but resistant to ticarcillin/clavulanic acid. No carbapenem resistance genes were detected. One transconjugant was successfully obtained by transconjugation assay. MLST analysis showed that a total of four sequence types were identified, including ST131 (two strains), ST43 (one strain),ST69 (one strain) and ST349(one strain). Conclusion Only 1.6% of all E.coli strains isolated in Dongyang area of Zhejiang Province carry mcr-1 gene,indicating that there is no epidemic of mcr-1 gene-positive E.coli infection. The coexistence of mcr-1 gene and β-lactamase resistance genes in E.coli strains isolated in Dongyang suggests that local clinicians should avoid antibiotic abuse to prevent the spread of drug-resistant E.coli.

Undifferentiated connective tissue disease (CTD) usually refers to patients who are presented with certain symptoms and signs related to CTD, and positive serological evidence of autoimmune diseases but don′t fulfill any of the classification criteria for a certain CTD. Mixed CTD refers to patients who are presented with one or more clinical manifestations such as hand swelling, synovitis, myositis, Raynaud′s phenomenon, and acrosclerosis. Patients with mixed CTD always have high-titer anti-nuclear antibodies (ANA) of speckled pattern and high-titer anti-U 1 ribonuclear protein (RNP) antibody in serum, while with negative anti-Sm antibody. The update of diagnosis and treatment of undifferentiated CTD and mixed CTD lags behind other established CTD. There is a lack of evidence from randomized controlled trials or guidelines/recommendations on the treatment of undifferentiated CTD or mixed CTD. At present, the conventional therapy is mainly adopted according to the specific clinical manifestations of the disease. The standardized diagnosis and treatment of undifferentiated CTD and mixed CTD were drafted by the Chinese Rheumatology Association based on the previous guidelines and the progress of available evidence, so as to improve the management of these patients in China.