Objective To study the diagnostic value in examination of cat scratch disease with CT.Methods The features of contrast enhanced CT scans in 3 cases with cat scratch disease proved by pathology and operation were analysed.Results Three cases all had clear scratches and contact history,lymph node enlargement along the regional lymphatic drainage was seen and presented as circular enhancementon CT.Conclusion To untypical patients with cat-scratch disease the contrast enhanced CT scans can show the local lymphatic pathological change and may help to comfirm the diagnosis.

Objective To evaluate the diagnostic efficiency of multi-parameter magnetic resonance imaging of routine MRI,including dynamic contrast-enhanced MRI(DCE-MRI)and diffusion weighed imaging(DWI)in diagnosis of breast cancer.Methods The results of routine MRI (morphology,boundary and,edge and Internal enhancement),DCE-MRI (1,2,3 minutes early enhanced rate and TIC curve)and DWI (b=500,800 s/mm2 )were analyzed in 43 cases of malignant breast lesions and 54 benign lesions,with pathology results asthe gold standard,the sensitivity,specificity,accuracy,positive predictive value,negative predictive value and Kappa value had been respectively gained,and Logistic regression analysis were used for statistical analysis.Results The specificity of edge (burr sign) was 96.3%,the sensitivity and specificity of TIC curve,ADC values (b=500,800 s/mm2 )were respectively 97.67%,83.33%,81.4%, 90.74%,88.37%,92.59%.Edge (burr sign)and ADC values (b=800 s/mm2 )had a statistic difference in the diagnosis of benign and malignant lesions (P<0.05),TIC curve had a significant statistic difference (P<0.01).Conclusion Edge (burr sign),TIC curve,ADC values (b=800 s/mm2 )had excellent diagnostic efficiency in the diagnosis of benign and malignant lesions.

Objective To develop and identify monoclonal antibodies (McAbs) against adult worm of Angiostrongylus cantonensis and observe its applicability. Methods BALB/c mice were immunized with soluble antigen of adult worms of A.cantonensis. The spleen cells of immunized mice were fused with myeloma cell, and the hybridoma secreting high titer of McAbs with high specificity was screened. By using the McAbs, serum of angiostrongyliasis patient and sera of the rats infected with A.cantonensis were detected by Western blotting and double antibody sandwich ELISA respectively. Results Three McAbs were established (2A2,3F1,4H2), which all showed no cross reaction with antigens of Schistosoma japonicum, Paragonimus westermani, Cysticercus cellulosae and Trichinella spiralis. Western blotting analysis demonstrated that the three McAbs recognized a Mr 15 000 soluble antigen of adult worm of A.cantonensis and recognized the Mr 24 000 and Mr 15 000 circulating antigens from the serum of angiostrongyliasis patient. The double antibody sandwich ELISA detection showed a positive rate of 76^5%. Conclusion Three hybridoma cell lines against adult worm of A.cantonensis have been established which secret high titer of McAbs with high specificity and seem promising in detecting the circulating antigen of the angiostrongyliasis patient.

Objective To clone and express prokaryotic recombinant plasmid of nucleoside triphosphate hydrolase (NTPase) gene of Toxoplasma gondii, and analyze its antigenicity. Method NTPase gene was amplified by PCR from RH strain of T. gondii and cloned into pGEM-T Easy vector. Positive clones were screened and identified by BglⅡ, HindⅢ digestion and sequenced. The target gene was then subcloned into prokaryotic expression vector pBAD-HisB and transformed into E. coli BL21(DE3). The expressed recombinant protein was purified with Ni-NTA agarose and further analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western blotting. Results NTPase-Ⅱ gene was specifically amplified, and the homology of DNA sequence was 100% to that in the GenBank. SDS-PAGE showed that the recombinant NTPase protein with correct molecular weight was expressed highly in E.coli BL21(DE3). Western blotting testified that the purified recombinant protein could be specifically recognized by mouse serum immunized with T. gondii and mouse anti-recombinant protein serum. Conclusion The NTPase-Ⅱ gene has been cloned and expressed in E.coli BL21(DE3), and the purified protein of NTPase-Ⅱ gene displays a specific antigenicity.

Objective To analyze the genetic characteristics of the complete genome of a human echovirus 30 (Echo30) KM/A363/09 strain isolated in Yunnan, China in 2009.Methods Primers specif-ic for Echo30 were designed .The extracted RNA was amplified by using RT-PCR.Seven fragments covering the complete viral genome were sequenced and the complete sequences were aligned with other sequences of enterovirus reference strains downloaded from Genbank . By using Mega5.1, Geneious, RDP3 and SimPlot3.5.1 softwares, the phylogenetic and recombination analysis were carried out .Results The com-plete nucleotide sequence of KM/A363/09 isolate was 7425 bp in length, encoding 2194 amino acids.KM/A363/09 isolate was highly similar with Bastianni prototype strain showing the homology of 81.2%in nucle-otide and 95.8%in amino acid.The eight Echo30 isolates shared 81.2%-88.6%homologies in nucleotide sequences and 95.8%-97.8%in amino acid sequences .Phylogenetic analysis showed that the KM/A363/09 strain belonged to one clade of Echo 30 in China.The genetic recombination of KM/A363/09 isolate was detected in the non-structural region .Conclusion KM/A363/09 isolate belongs to one clade of Echo 30 in China indicating that the evolution of Echo 30 has occurred in China .

OBJECTIVE:To investigate the inhibitory effect of siRNA expression vector inhibiting human insulin-like growth factor 2(IGF2)gene on the proliferation of hepatoma cell line Huh-7. METHODS:siRNA expression vector pGL3-hAFP-hTERT-siRNA3(“siRNA3”)which inhibited IGF2 gene by dual promoter regulation of recombinant human alpha-foetoprotein(hAFP)and human telomerase reverse transcriptase(hTERT)was transfected into the Huh-7 cell and normal hepatocyte L-02,and then a nega-tive control group(vector pGL3-hAFP-hTERT)and a blank control group were set up. IGF2 mRNA expression was detected by re-al-time fluorescent quantitative polymerase chain reaction 48 h after transfection into the cells in all groups;the activity of the cells by the microplate reader 0,24,48 and 72 h thereafter;and the cell cycle and apoptosis by the flow cytometer 48 h thereafter,and the changes in the protein levels of IGF2,PCNA,Cyclin E2,Cyclin D2,Cdc2 and Bcl-2 in the cell were detected by Western blot. RESULTS:Compared with the negative control group and blank control group,IGF2 mRNA expression in the Huh-7 cell transfected with siRNA3 was obviously weaker;at 48 and 72 h after transfection,the activity of Huh-7 cell signigicantly reduced, Huh-7 cells at G1 phase obviously increased and those at S phase markedly decreased;the occurrence of early,late and total apopto-sis in Huh-7 cells apparently increased,and the protein expression of IGF2,PCNA,Cyclin E2,Cyclin D2,Cdc2 and Bcl-2 in cells significantly weakened,with statistically significance(P<0.01 or P<0.05). No obvious change in the above-mentioned index-es could be found in L-02 cells transfected with siRNA3 expression vector (P>0.05). CONCLUSIONS:siRNA which inhibited IGF2 gene by dual promoter regulation of recombinant hAFP and hTERT can specially inhibit IGF2 gene expression and the prolifer-ation of Huh-7 cells,which may be involved with down-regulated protein expression of cell proliferation-associated gene PCNA, cell cycle control-associated genes Cyclin E2,Cyclin D2 and Cdc2 and apoptosis regulation-associated gene Bcl-2 as a result of down-regulated IGF2 mRNA expression and protein expres-sion.

Objective To analyze the complete genome sequence of a coxsackievirus B 3 (CVB3) strain KM06/2009 and its genetic variation , evolution and cardiovirulence .Methods Eight clones with overlapped gene fragments covering the complete viral genome ( excluding the poly-A tail) were obtained by RT-PCR and sequenced .The nucleotide ( nt ) and amino acid ( aa ) sequences of the eight clones were aligned with sequences of other known CVB 3 clinical strains .Phylogenetic and pairwise alignment analyses based on the genome and complete VP 1 regions were conducted by using Mega 4.1,RDP3 and SimPlot3.5.1 softwares.The RNA secondary structure of CVB3 stem loopⅡ (SLⅡ) was determined by using Mfold web server.Results The complete genome of CVB3 strain KM06/2009 was 7401 nt in length, consisting of 743 nt and 100 nt on 5′untranslated region (UTR) and 3′UTR,respectively.The entire open reading frame contained 6558 nt, encoding a 2185 aa polyprotein.There was no nucleotide deletion or insertion in the coding region,but some changes of amino acid were unique .KM06/2009 strain showed 81.4%and 95.7%identities in nucleotide and amino acid sequences respectively as compared with CVB 3 prototype Nancy strain.It shared 88.4%-98.1%nucleotide and 98.1%-99.4%amino acid homology with the other Chinese clinical strains isolated at the same period .KM06/2009 strain and CVB3 GA strain without cardiovirulence shared 80.7%homology in nucleotide and 96.4% in amino acid.The phylogenetic analysis indicated that the significant spatial and temporal clustering was detected in CVB 3 isolate.CVB3 KM06/2009 strain showed a strong cardiovirulence tendency as indicated by the RNA secondary structure of CVB 3 SL Ⅱ. Conclusion CVB3 KM06/2009 isolate showed a strong cardiovirulence tendency in comparison with other CVB3 clinical isolates based on the bioinformatics analysis .

Objective To explore the relationship between the osteonecrotic volume (lesion size), angle and the index of necrotic extent on the femoral head. Methods Fifty-one hips in 39 patients with non-traumatic avascular osteonecrosis on the femoral head were divided into 12 equal segments from the head to the neck junction (a turning point of the spherical curve of the head) with whole hip displacement, each with 30 degrees on a coronal plane of weight-bearing surface. The osteonecrotic angle of the arc of each necrotic segment from the center of the femoral head was measured at the point of one- to 12-o'clock on imaging of two dimensional reconstruction of computerized tomography (CT) scans and anteroposterior and lateral radiographs. Necrotic volume on each segment of the femoral head was calculated with fluid displacement method. The index of osteonecrotic extent on the femoral head was calculated using direct anatomical measurements. Results Osteoneerotic volume on the femoral head at the point of 12- to one-o'clock on coronal plane was (74. 5 ± 7.4)% of the sphere equivalent of the whole femoral head, which was positively correlated to its necrotic angle [ (41.9±8. 3) degrees] at the point of one-o'clock on the plane, with a coefficient of correlation of 0. 843, and that at the point of one- to two-o'clock on the plane was (73.7 ±0. 4) %, which was positively correlated to its necrotic angle [ (41.9 ± 1.8) degrees] at the point of two-o'clock, with a coefficient of correlation of 0. 543. Osteonecrotic volume on the point of 11- to 12-o'cleck was (83.6±8.6)%, and the necrotic angle at the point of 12-o'clock was (44. 9±3.9) degrees, which were positively correlated each other, with a coefficient of correlation of 0. 701 (P <0. 01 ). Osteonecrotic volume on the femoral head was positively correlated to its necrotic angle , modified index of necrotic extent, index of necrotic extent and Kerboul conjugated necrotic angle, with coefficients of correlation of 0. 798, 0. 701, 0. 377 and 0. 398 ( P < 0. 01 ), respectively, at the point of one o' clock. Conclusions Measurements of osteonecrotic volume was positively correlated to the index of necrotic extent and necrotic angle on the femoral head, respectively. Necrotic angle on the plane at 30-degree of the trochanter on the femoral head can well reflect its necrotic volume.

Objective:To investigate the serum 25-hydroxyvitamin D [25(OH) D] levels in elderly patients with hip and vertebral compression fractures (VCF).Method:Ninety patients (58 males and 32 females) aged over 60 years with hip fracture and 120 patients (88 males and 32 females) aged over 60 years with VCF admitted in the Aviation General Hospital from January 2017 to June 2019 were enrolled. Serum 25(OH)D levels were measured.Results:Serum level of 25 (OH) D in hip fracture patients was (9.0±6.8) μg/L, the 25 (OH) D level was lower than the normal value(<19.0 μg/L)in 79 patients and<3 μg/L in 24 patients. The level of 25(OH)D in VCF patients was (16.7±10.6) μg/L, the 25 (OH) D level was<19.0 μg/L in 78 patients (65.0%) and <3 μg/L in 10 patients (8.3%). The low level of 25(OH)D was negatively correlated with age in two groups ( r=-0.367, P=0.01; r=-0.313, P=0.04). The mean level of 25 (OH) D in the hip fracture group was lower than that in the VCF group ( t=5.960, P<0.01), and the low 25(OH)D rate in the former group was significantly higher than that in the latter group (χ 2=14.14, P<0.01; χ 2=12.74, P<0.01). The 25(OH)D value of female VCF patients was (14.5±8.8) μg/L, which was significantly lower than that of male patients (22.5±12.9) μg/L ( t=3.882, P<0.01).Among hip fracture patients, the 25(OH)D level in patients with fracture history was (8.3±6.9) μg/L, which was significantly lower than that of patients without fracture history (10.8±6.9) μg/L, and the difference was statistically significamt ( t=2.123, P=0.04). The serum osteocalcin level was (20.5±19.8) μg/L in patients with fracture history, which was significantly higher than that in patients without fracture history [(10.6±5.4) μg/L, t=3.245, P<0.01]. Conclusion:Elderly patients with new hip fractures have more severely low vitamin D level than patients with new VCF, and patients with previous fracture history have lower vitamin D levels than patients without fracture history.

BACKGROUND: Osteoporosis (OP) has become a major public health issue, threatening the bone health of middle-aged and elderly people from all around the world. Changes in the gut microbiota (GM) are correlated with the maintenance of bone mass and bone quality. However, research results in this field remain highly controversial, and no systematic review or meta-analysis of the relationship between GM and OP has been conducted. This paper addresses this shortcoming, focusing on the difference in the GM abundance between OP patients and healthy controls based on previous 16S ribosomal RNA (rRNA) gene sequencing results, in order to provide new clinical reference information for future customized prevention and treatment options of OP. METHODS: According to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA), we comprehensively searched the databases of PubMed, Web of Science, Embase, Cochrane Library, and China National Knowledge Infrastructure (CNKI). In addition, we applied the R programming language version 4.0.3 and Stata 15.1 software for data analysis. We also implemented the Newcastle-Ottawa Scale (NOS), funnel plot analysis, sensitivity analysis, Egger's test, and Begg's test to assess the risk of bias. RESULTS: This research ultimately considered 12 studies, which included the fecal GM data of 2033 people (604 with OP and 1429 healthy controls). In the included research papers, it was observed that the relative abundance of Lactobacillus and Ruminococcus increased in the OP group, while the relative abundance for Bacteroides of Bacteroidetes increased (except for Ireland). Meanwhile, Firmicutes, Blautia, Alistipes, Megamonas, and Anaerostipes showed reduced relative abundance in Chinese studies. In the linear discriminant analysis Effect Size (LEfSe) analysis, certain bacteria showed statistically significant results consistently across different studies. CONCLUSIONS: This observational meta-analysis revealed that changes in the GM were correlated with OP, and variations in some advantageous GM might involve regional differences.
Middle Aged ; Aged ; Humans ; Gastrointestinal Microbiome/genetics* ; RNA, Ribosomal, 16S/genetics* ; Genes, rRNA ; Osteoporosis ; Feces