ObjectiveTo study the dynamic express of CD57 on T cell of PBMC and clinical significance in acute HIV infection.MethodsSeventeen patients with acute HIV infection were enrolled study randomly diagnosed from 2006.11 to 2009.12 and 15 healthy donors as control group.The PBMCs from 1th,3th and 6th during acute infection were collected.The proportion of CD3+CD57+T lymphocytes,CD3+ CD8+CD57 +T lymphocytes and CD3 + CD4 + CD57 + T lymphocytes were evaluated by flow cytometric analysis with three or double color staining.The relationship between the proportion of CD57+ T phenotypes and virus load and CD4+T cells count was analyzed.ResultsThe proportion of CD57+T lymphocytes in PMBC in 1th,3th and 6th during acute HIV acute was 15.24％ ±1.49％,13.51％ ±2.45％ and 14.65％ ±1.83％,respectively,and was higher than normal control group and the difference was significantly(P＜0.0001 ).The proportion of CD8+ CD57+T lymphocytes was 7.79％ ±2.10％ and 9.88％ ±2.36％ in 1th and 3th month during acute infection,respectively.The proportion of CD8+ CD57+T lymphocytes in 1th and 3th month during acute infection were positive relationship with virus load in corresponding time,and R2 was 0.3700 and 0.3768,and P value was 0.0096 and 0.0088,respectively.The proportion of CD8+CD57+T lymphocytes in the 1th and 3th month during acute infection was negative relationship with CD4+T lymphocytes count.The R2 was 0.3768 and 0.4235,and P value was 0.0215 and 0.0017,respectively.In 6 rapid progressors and 11 no rapid progressors on the 1th month after HIV infection,CD8+ CD57+T lymphocytes percentage was 11.20％±2.21％ and 6.16％ ±1.09％,respectively,and CD4+CD57+T lymphocytes percentage 2.79％ ±0.31％and 1.40％ ±0.30％,respectively.Both CD8+CD57+T and CD4+CD57+T in rapid progressors were higher than no rapid progressors,and P value was 0.0338 and 0.0106,respectively.ConclusionCD57+ T lymphocytes percentage in peripheral blood increase in acute HIV infection patients,in which the increasing CD8+CD57+T lymphocyte may mirror the dynamic of HIV replication and CD4+T cell count.The CD57 high express on T lymphocyte on the early HIV acute infection predicts rapid progression.

Objective To investigate the underlying mechanisms of Miller-Fisher syndrome (MFS) and Bickerstaff' s brainstem encephalitis (BBE) by studying their clinical and electrophysiological characteristics.Methods The clinical and electrophysiological characteristics of 13 MFS and 7 BBE cases in Peking Union Medical College Hospital between 2000 and 2011 were retrospectively analyzed.The electrophysiological parameters included sensory and motor nerve conduction,electromyography,F wave,sympathetic skin response and brainstem auditory evoked potential and blink reflex.Results MFS and BBE had similar clinical characteristics:respiratory symptoms were the most common infectious symptoms before disease onset; Ophthalmoplegia,facial palsy and bulbar symptoms were common; They both had cerebrospinal fluid albuminocytological dissociation and positive serum anti-GQ1b antibody.However,BBE had more central nervous system lesion signs clinically such as conscious disturbance,positive Babinski' s sign and central facial palsy.Electrophysiologically,MFS and BBE also had similar electrophysiological features:sensory nerve abnormality ratios were 6/13,2/7 respectively,with prominently reduced sensory nerve active potential amplitude,normal or slightly slowed sensory conduction velocity; Motor nerves abnormality ratios were 2/13,1/7 respectively,with slightly prolonged distal motor latency and normal compound muscle action potential; Electromyography abnormality ratios were 1/7,0/4 respectively; F wave frequency abnormality ratios were 3/13,5/7 respectively,and in some cases,F wave frequency would restore; Sympathetic skin response abnormality ratios were 1/2,1/3 respectively; Blink reflex abnormalityratios were 1/2,1/1 respectively,with central involvement in BBE; Brainstem auditory evoked potential abnormality ratios were 3/5,1/4 respectively,with wave Ⅰ latency or amplitude abnormality.Conclusion The similarities of clinical and electrophysiological features suggest that MFS and BBE have the same mechanism and they form a continuous spectrum with variable central nervous system and peripheral nervous system involvement.

Objective To investigate the risk factors for acute calf muscular venous thrombosis(CMVT)in elderly patients with hip fractures and evaluate the diagnostic value of thrombotic risk index for CMVT.Methods The blood samples from elderly emergency admission patients with traumatic hip fractures were prospectively collected,and platelet count(PLT),plateletcrit(PCT),coagula-tion markers such as thrombin-activatable fibrinolysis inhibitor(TAFI),D-dimer,and fibrinogen(Fib),and biochemical markers,including serum total protein(TP),globulin(Glob),creatine kinase(CK),triglyceride(TG),and lactate dehydrogenase(LDH),were detected.The differences in these markers between the CMVT group,deep calf venous thrombosis(DCVT)group and non-deep venous thrombosis(DVT)group were compared.The multivariate logistic regression analysis was used to identify independent risk fac-tors for CMVT,and a thrombotic risk index was generated to evaluate its diagnostic value in CMVT.Results Compared with the non-DVT group,the levels of TAFI,Fib,TP,CK,TG,PCT,and LDH in the CMVT group were significantly reduced(P<0.05),while the level of D-dimer increased but the difference was not statistically significant.The level of TP in the DCVT group was significantly lower than that in the non-DVT group(P<0.05).Multivariate analysis showed that age,LDH,and Fib were independent risk factors for CMVT.The area under the receiver operating characteristics(ROC)curve of the thrombotic risk index in diagnosing CMVT was 0.718(P<0.001).Conclusion Age,LDH,and Fib are the independent risk factors of CMVT in elderly patients with hip fractures.The thrombotic risk index has a high diagnostic value in CMVT.

OBJECTIVETo explore the distribution characteristics of the types of M protein gene (emm) in group A streptococcus (GAS) isolated from children in Beijing in the year 2011.

METHODSDuring May to July in 2011, a total of 3315 patients who were diagnosed scarlet fever or pharyngeal infection by doctors in pediatric outpatient and emergency units of 36 hospitals, were selected as subjects. Their throat swab samples were collected and isolated the strains of GAS. Gene emm was then amplified and sequenced by PCR method, and the differences in types of gene emm between different populations and diseases were compared.

RESULTSA total of 633 strains of GAS were isolated from the 3315 throat swab samples, 610 strains out of which were gene emm positive and were recruited in the study. Out of the 610 recruited strains, 448 (73.4%) were isolated from scarlet fever patients, the other 162 (26.6%) were isolated from pharyngeal infection patients; 397 (65.1%) were from urban, the other 213 (34.9%) were from suburb; 240 (39.4%) were from patients aging between 1 - 5 years old, the other 369(60.6%) were from patients aging 6 - 18 years old. A total of 8 types of gene emm (scarlet fever: 6 types, pharyngeal infection: 4 types) and 21 subtypes of gene emn (scarlet fever: 16 subtypes, pharyngeal infection: 10 subtypes) were identified. Three new subtypes were found in the study, naming emm1.63, emm12.62 and st5144.20. Among them, emm1.63 was found both in scarlet fever and pharyngeal infection patients, while emm12.62 and st5144. 20 were only found in pharyngeal infection patients. Among all the types of gene-emm, emm12 accounted for the highest percentage as 80.5% (491/610) and then followed by emm1 (18.0% (110/610)). Among all the subtypes, the dominant subtype was emm12.00, accounting for 69.0% (421/610), following by emm1.00 (16.9% (103/610)) and emm12.19 (6.1% (37/610)). All the above types and subtypes of gene emm were the most prevalent strains in scarlet fever patients and pharyngeal infection patients. Significant differences in the distribution of prevalent strains were observed among various aging patients and regions. The constituent ratios of emm1, emm1.00 and emm12.19 were higher in patients from suburb (emm1: 22.1% (47/213), emm1.00: 19.2% (40/213), emm12.19: 8.0% (17/213)) than those in urban areas (emm1: 15.9% (63/397), emm1.00: 15.6% (62/397), emm12.19: 5.0% (20/397)). The difference showed statistical significance (P < 0.05). The constituent ratio of emm1.00 was higher among patients aging 6-18 years old (19.2% (71/369)) than those aging 1 - 5 years old (13.3% (32/240)). The difference also showed statistical significance (χ(2) = 8.45, P < 0.05).

CONCLUSIONAmong the types of gene emm in GAS isolated from children in Beijing in year 2011, the most prevalent two were emm12 and emm1, and the most prevalent emm subtypes were emm12.00, emm1.00 and emm12.19. A significant difference in their distribution between various aging patients and isolated places can be obviously found.

Adolescent ; Antigens, Bacterial ; classification ; genetics ; Bacterial Outer Membrane Proteins ; classification ; genetics ; Carrier Proteins ; classification ; genetics ; Child ; Child, Preschool ; China ; Female ; Genes, Bacterial ; Genotype ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Streptococcus pyogenes ; genetics ; isolation & purification

TGFβ/smad pathway is recognized as an important signal pathway to promote the pathogenesis of atherosclerosis (AS). Peroxisome proliferator-activated receptor γ (PPARγ) activation is considered to be important in modulating AS. Herein, we investigated the regulation of PPARγ on c-Ski, the repressor of TGFβ/smad pathway, in rat AS model and cultured vascular smooth muscle cells (VSMCs). c-Ski mRNA and protein expression were detected by real-time PCR and Western blot, respectively, in vivo and in vitro with treatment of PPARγ agonist rosiglitazone and antagonist GW9662. The proliferation and collagen secretion of VSMCs after c-Ski transfection were investigated. The underlying mechanism was further investigated by online program NUBIScan and luciferase reporter gene analysis. Results showed that both mRNA and protein expressions of c-Ski in the AS lesions was down-regulated in vivo, while in cultured VSMCs, c-Ski transfection significantly suppressed the proliferation and collagen secretion of rat VSMCs. Rosiglitazone significantly up-regulated mRNA and protein levels of c-Ski in VSMCs, which could be blocked by GW9662. Online NUBIScan analysis suggested possible PPARγ binding sites in the promoter region of c-Ski. In addition, luciferase activity of c-Ski reporter gene was also increased obviously in the presence of rosiglitazone. These results indicate that c-Ski is one of the newly found target genes of PPARγ and thus involved in the anti-AS effect of PPARγ.
Anilides ; pharmacology ; Animals ; Atherosclerosis ; physiopathology ; Cells, Cultured ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; PPAR gamma ; agonists ; antagonists & inhibitors ; physiology ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Repressor Proteins ; genetics ; metabolism ; Signal Transduction ; Smad Proteins ; metabolism ; Thiazolidinediones ; pharmacology ; Transforming Growth Factor beta ; metabolism ; Up-Regulation

Objective To establish a novel convenient loop?mediated isothermal amplification(LAMP)method with the unique genes coding Plasmodium helical interspersed sub?telomeric superfamily(PHIST)for the rapid molecular diagnosis of P. falciparum. Methods The unique genes coding PHIST with high expression mRNA profile during the ring form or schizont period of P. falciparum were screened and selected from the PlasmoDB database. The LAMP primers of targeted genes were de?signed by the online software(PrimerExplorer V4). The LAMP assay was executed by the color?displaying method with SYBR Green. The dried blood spots of P. falciparum from clinical isolates were collected and the genomic DNA(gDNA)was extracted. For evaluation of sensitivity,the gDNA was diluted to four gradients(10?1,10?2,10?3,and 10?4). For assessment of specificity, the gDNA(s)of P. vivax,P. yoelii,Taenia saginata,and Schistosoma japonicum were also extracted. Results Totally,61 P. falciparum unique genes coding PHIST were found. The PF3D7_1372300 with high expression value during the ring form and PF3D7_1401600 with high expression value during the schizont period were selected for LAMP assay. The lowest detectable lim?its of PF3D7_1372300 and PF3D7_1401600 were 130.5 parasite/μl and 1 305.3 parasite/μl,respectively. Specific tests showed the amplified products of P. falciparum was positive and all the others including P. vivax,P. yoelii,T. saginata,and S. japoni?cum were negative. Conclusions The established LAMP method with PF3D7_1372300 gene is sensitive,specific,simple and useful. It can be applied to the field investigation and clinical diagnosis for falciparum malaria.

OBJECTIVETo analyze the phenotype and genotype of a family with inherited dysfibrinogenemia.

METHODSLaboratory tests including activated particle thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), and the activity of protein C (PC), protein S(PS) and antithrombin (AT) were conducted in the proband and 4 family members. The activity and antigen of fibrinogen in plasma were measured by functional and immunoturbidimetry assay, respectively. All the exons and exon-intron boundaries of the three fibrinogen genes were analyzed by direct sequencing.

RESULTSThe proband had normal APTT and PT, but prolonged TT. Her plasma fibrinogen levels were extremely reduced, which was also found in her mother. The sequencing results of the proband revealed heterozygous g.5678 G>A in the exon 8 of FGG gene originating from her mother, which caused Arg275His missense mutation.

CONCLUSIONDysfibrinogenemia in the family is caused by Arg275His in the beta chain of fibrinogen and it is the first report on a Chinese family with inherited dysfibrinogenemia.

Adult ; Afibrinogenemia ; blood ; genetics ; Amino Acid Substitution ; Arginine ; genetics ; DNA Mutational Analysis ; Female ; Fibrinogen ; genetics ; metabolism ; Histidine ; genetics ; Humans ; Male ; Pedigree ; Phenotype

OBJECTIVETo investigate the effect of rabbit saphenous and sciatic nerve homogenates on the proliferation and calcification of rabbit osteoblasts in vitro.

METHODThe saphenous nerves (sensory nerves) and the muscular branches of the sciatic nerve (motor nerve) were collected from 48 New Zealand white rabbits to prepare the nerve tissue homogenates. Bone marrow mesenchymal stem cells (MSCs) were isolated from the rabbits and cultured in vitro, and after 14 days of routine osteogenic induction, the resultant osteoblasts were identified by immunohistochemistry, alkaline phosphatase (ALP) and Alizarin red S staining. The osteoblasts were then incubated in the induction medium containing the saphenous (sensory nerve group) or sciatic homogenates (motor nerve group), with the cells in the dexamethasone-containing, dexamethasone-free osteogenic induction medium and control medium as the control. The proliferation, total protein and ALP activity of the osteoblasts were measured every other day until the 8th day, and Alizarin red S staining was used for quantitative analysis of calcification of the cells after two weeks.

RESULTSThe application of the saphenous nerve homogenates significantly promoted cell proliferation, total protein and ALP activity (P<0.01, P<0.05 and P<0.05), while exposure of the osteoblasts to dexamethasone inhibited the cell proliferation (P<0.001). Compared to dexamethasone-free group, the saphenous homogenates enhanced the mineralization of the osteoblasts (P<0.001).

CONCLUSIONSaphenous nerve homogenates significantly promotes the proliferation, differentiation, ALP activity and mineralization of rabbit osteoblasts, but sciatic nerve homogenates do not show osteogenic effects on the cells.

Animals ; Bone Marrow Cells ; cytology ; Calcification, Physiologic ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; physiology ; Rabbits ; Sciatic Nerve ; chemistry ; Sensory Receptor Cells ; chemistry ; Tissue Extracts ; pharmacology

Chinese patent medicine (CPM) is an important part of traditional and Chinese medicine (TCM). Its quality has direct impact on the safety and effectiveness of clinical use. The quality standard is the pivotal approach to guarantee the quality of CPM. Due to the complex material basis, multitudinous quality influencing factors and unveiled active ingredients, dose-effect relationship and action mechanism, the investigation on quality standard faces many difficulties. This paper surveys the current quality status of CPM and the general situation of CPM standards. At present, the dosing problem has the crucial impact on the quality of CPM. The current quality standard system of CPM is confirmed and the limitations are indicated. Based on the above analysis, the principles and considerations on investigation of quality standard are proposed as follows: ① Adhere to safety as the bottom line, strengthen the risk-control ability of the standard of CPM; ② Adhere to theory of TCM and comprehensive quality, improve the integrative control level of the CPM standard; ③ Emphasize technological development and innovation, promote the quality control competence of CPM standard; ④ Facilitate planning and coordination, optimize the management of the CPM standard system; ⑤ Reinforce investigation on evaluation method, develop grade evaluation standard, accelerate high-quality development of CPM. Finally, the future perspective on investigation of CPM quality standard is prospected.

Objective:To investigate the correlation between morphological typing and monoclonality of bone marrow plasma cells,and explore the diagnostic value of plasma cell morphological typing for high-risk smoldering multiple myeloma(HR-SMM).Methods:The correlation between the morphological characteristics and the monoclonality of bone marrow plasma cells was analyzed in 84 patients with HR-SMM who treated in our hospital.The consistency of morphologically abnormal bone marrow plasma cells with serum free light chain(sFLC)ratio,next-generation sequencing(NGS)detection results,and its correlation with monoclonal plasma cells detected by flow cytometry(FCM)were further verified.The immunoglobulin types and levels of non-involved immunoglobulins in serum of the patients were detected,and the distribution of plasma cell clusters in patients with different disease was observed.Results:The mean percentage of mature plasma cells were decreased successively in the order of reactive plasmacytosis(RP)group,monoclonal gammopathy of undetermined significance(MGUS)group,smoldering multiple myeloma(SMM)group,HR-SMM group and multiple myeloma(MM)group;while the mean percentage of immature,primitive,reticular and flaming plasma cells were increased successively in the order of RP group,MGUS group,SMM group,and HR-SMM group,and the difference between any two groups was statistically significant(P＜0.05).The average proportion of abnormal plasma cells in the bone marrow of HR-SMM patients was 96.2％of the total plasma cells.The proportion of abnormal plasma cells were in good agreement with the sFLC ratio and the results of NGS detection in HR-SMM patients(kappa=0.879 and kappa=0.891,both＞0.75),and showed good correlation with the monoclonal plasma cells with immunophenotype of CD45-/CD38+/CD138+/CD56+/CD19-(γ=0.825).The levels of non-involved immunoglobulin in IgG,IgA and IgM type HR-SMM patients were all decreased by more than 25％compared with the normal reference range,and the differences were statistically significant(P＜0.05).There was no significant difference in the distribution ratio of plasma cell clusters among different disease groups(P＞0.05).Conclusion:In HR-SMM patients,the immature,primitive,reticular and flaming plasma cells in bone marrow are considered as abnormal plasma cells,and they are correlated with monoclonal plasma cells.The proportion of abnormal plasma cells in total plasma cells of bone marrow and the reduction extent of non-involved immunoglobulin level in patients have certain reference value for the diagnosis of HR-SMM.