ObjectiveThis study leverages structural data from antigen-antibody complexes of the influenza A virus neuraminidase (NA) protein to investigate the spatial recognition relationship between the antigenic epitopes and antibody paratopes. MethodsStructural data on NA protein antigen-antibody complexes were comprehensively collected from the SAbDab database, and processed to obtain the amino acid sequences and spatial distribution information on antigenic epitopes and corresponding antibody paratopes. Statistical analysis was conducted on the antibody sequences, frequency of use of genes, amino acid preferences, and the lengths of complementarity determining regions (CDR). Epitope hotspots for antibody binding were analyzed, and the spatial structural similarity of antibody paratopes was calculated and subjected to clustering, which allowed for a comprehensively exploration of the spatial recognition relationship between antigenic epitopes and antibodies. The specificity of antibodies targeting different antigenic epitope clusters was further validated through bio-layer interferometry (BLI) experiments. ResultsThe collected data revealed that the antigen-antibody complex structure data of influenza A virus NA protein in SAbDab database were mainly from H3N2, H7N9 and H1N1 subtypes. The hotspot regions of antigen epitopes were primarily located around the catalytic active site. The antibodies used for structural analysis were primarily derived from human and murine sources. Among murine antibodies, the most frequently used V-J gene combination was IGHV1-12*01/IGHJ2*01, while for human antibodies, the most common combination was IGHV1-69*01/IGHJ6*01. There were significant differences in the lengths and usage preferences of heavy chain CDR amino acids between antibodies that bind within the catalytic active site and those that bind to regions outside the catalytic active site. The results revealed that structurally similar antibodies could recognize the same epitopes, indicating a specific spatial recognition between antibody and antigen epitopes. Structural overlap in the binding regions was observed for antibodies with similar paratope structures, and the competitive binding of these antibodies to the epitope was confirmed through BLI experiments. ConclusionThe antigen epitopes of NA protein mainly ditributed around the catalytic active site and its surrounding loops. Spatial complementarity and electrostatic interactions play crucial roles in the recognition and binding of antibodies to antigenic epitopes in the catalytic region. There existed a spatial recognition relationship between antigens and antibodies that was independent of the uniqueness of antibody sequences, which means that antibodies with different sequences could potentially form similar local spatial structures and recognize the same epitopes.

In this study, we investigated the mechanism of crude extract of Psammosilene tunicoides(CEPT) in the treatment of rheumatoid arthritis(RA) based on the Nod-like receptor protein 3(NLRP3) inflammasome. The collagen-induced arthritis(CIA) mouse model was established. On day 32 after the primary immunization, according to the arthritis score, the mice were randomly divided into model group, positive control(methotrexate) group, low-and high-dose CEPT groups, and normal group, with 10 mice in each group. According to the administration dose of each group, the mice were continuously administered for 21 days. Every four days during the administration, the paw edema degree, arthritis score, and spleen index of the mice were measured; histopathological examination was performed for the ankles of the mice; the contents of IL-1β and IL-18 in the serum were determined; the protein expression levels of NLRP3, caspase-1, and apoptosis-associated speck-like protein containing a CARD(ASC), as well as the mRNA expression levels of NLRP3 and caspase-1 in the ankle joints of the mice were detected. The results showed that compared with those in the model group, the mice in the positive control group and CEPT groups had significantly decreased the contents of IL-1β and IL-18 in the serum and spleen index(P<0.01), significantly lowered arthritis score and degree of paw edema(P<0.01), alleviated arthritic infiltration of the knee, and down-regulated protein and mRNA levels of NLRP3, ASC, and caspase-1 in the ankle joint(P<0.01). These results suggest that P. tunicoides may reduce the paw edema and arthritis score and alleviate the inflammatory response in CIA mice by inhibiting the expression of NLRP3. This study provides a basis for the study of immune regulation of P. tunicoides in RA.
Animals ; Arthritis, Experimental/genetics* ; Arthritis, Rheumatoid/genetics* ; Caspase 1/genetics* ; Inflammasomes/genetics* ; Mice ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics*

OBJECTIVETo investigate the prognostic value of F-FDG PET/CT scan quantization parameters, max standardized uptake value （SUVmax）, metabolic tumor volume （MTV）, total lesion glycolysis （TLG） and other clinical factors for prognostic evaluation of paticnts with diffuse large B-cell lymphoma （DLBCL）.

METHODSPET/CT scan and clinical data of a total of 65 newly diagnosed DLBCL patients who received Rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone （R-CHOP） chemotherapy as first-line treatment were analyzed. All patients received a PET/CT scan at diagnosis and an interim PET/CT after 2-4 circles of chemotherapies. The related parameters of SUVmax, MTV and TLG were acquired by analyzing and calculating the scan results. The receiver operating characteristic （ROC） curve was used to determine the optimal cut-off of parameters. Pearson chi-square test, Kaplan-Meier method and COX proportional hazard model were performed to analyze the prognostic value of PET/CT related parameters and clinical factors in progression-free survival （PFS）.

RESULTSAge, B symptom, Ann Arbor stage and extra-nodal involvement in major organs significantly related with PFS （P<0.05）, but the SUVmax didn't relalt with the prognosis. The cut-off values of MTV0, MTV1, TLG0 and TLG1 for disease recurrence or progression were 172.20cm , 4.32cm , 1043.33g and 14.07g. The lower MTV and TLG groups showed longer PFS significantly. In the multivariate Cox regression model, B symptoms, MTV1 and TLG1 were the independent prognostic risk factors.

CONCLUSIONMTV and TLG at baseline and in the interim and NCCN-IPI correlate with disease prognosis. SUVmax related parameters hare no significant relationship with prognosis. Besides MTV and TLG during treatment are the independent prognostic risk factors suggesting more predictive value than NCCN-IPI.

BACKGROUNDCongenital myasthenic syndromes (CMSs) are a group of clinically and genetically heterogeneous disorders caused by impaired neuromuscular transmission. The defect of AGRN was one of the causes of CMS through influencing the development and maintenance of neuromuscular transmission. However, CMS reports about this gene mutation were rare. Here, we report a novel homozygous missense mutation (c.5302G>C) of AGRN in a Chinese CMS pedigree.

METHODSWe performed a detailed clinical assessment of a Chinese family with three affected members. We screened for pathogenic mutations using a disease-related gene panel containing 519 genes associated with genetic myopathy (including 17 CMS genes).

RESULTSIn the family, the proband showed limb-girdle pattern of weakness with sparing of ocular, facial, bulbar, and respiratory muscles. Repetitive nerve stimulation showed a clear decrement of the compound muscle action potentials at 3 Hz only. Pathological analysis of the left tibialis anterior muscle showed predominance of type I fiber and the presence of scattered small angular fibers. The proband's two elder sisters shared a similar but more severe phenotype. By gene analysis, the same novel homozygous mutation (c.5302G>C, p. A1768P) of AGRN was identified in all three affected members, whereas the same heterozygous mutation was found in both parents, revealing an autosomal recessive transmission pattern. All patients showed beneficial responses to adrenergic agonists.

CONCLUSIONSThis study reports a Chinese pedigree in which all three children carried the same novel AGRN mutation have CMS only affecting limb-girdle muscle. These findings might expand the spectrum of mutation in AGRN and enrich the phenotype of CMS.

Background: Congenital myasthenic syndromes (CMSs) are a group of clinically and genetically heterogeneous disorders caused by impaired neuromuscular transmission. The defect of AGRN was one of the causes of CMS through influencing the development and maintenance of neuromuscular transmission. However, CMS reports about this gene mutation were rare. Here, we report a novel homozygous missense mutation (c.5302G>C) of AGRN in a Chinese CMS pedigree. Methods: We performed a detailed clinical assessment of a Chinese family with three affected members. We screened for pathogenic mutations using a disease?related gene panel containing 519 genes associated with genetic myopathy (including 17 CMS genes). Results: In the family, the proband showed limb?girdle pattern of weakness with sparing of ocular, facial, bulbar, and respiratory muscles. Repetitive nerve stimulation showed a clear decrement of the compound muscle action potentials at 3 Hz only. Pathological analysis of the left tibialis anterior muscle showed predominance of type I fiber and the presence of scattered small angular fibers. The proband's two elder sisters shared a similar but more severe phenotype. By gene analysis, the same novel homozygous mutation (c.5302G>C, p.A1768P) of AGRN was identified in all three affected members, whereas the same heterozygous mutation was found in both parents, revealing an autosomal recessive transmission pattern. All patients showed beneficial responses to adrenergic agonists. Conclusions: This study reports a Chinese pedigree in which all three children carried the same novel AGRN mutation have CMS only affecting limb?girdle muscle. These findings might expand the spectrum of mutation in AGRN and enrich the phenotype of CMS.

To study the antirheumatic substance of Loranthus parasiticus and observe the relationship between its in vivo distribution and meridian tropism in rats by establishing adjuvant arthritis models corresponding to effectiveness. All rats except the negative control group were injected with 0.1 mL Freund's complete adjuvant on the left foot. After 8 days, the rats in negative control group and model group were given with normal saline while the rats in positive control group were given with tripterygium glycosides suspension 10 mg•kg-1, and the rats in L. parasiticus treatment groups were given with high(10 g•kg ⁻¹), medium(5 g•kg ⁻¹) and low(2.5 g•kg ⁻¹) dose decoction for 21 days. The left rear ankle joint diameter of rats were measured every 7 days from the 9th day of modeling. On the 22nd day, eyeball blood of part rats in L. parasiticus high-dose group was taken at different time points, and then they were sacrificed to take heart, liver, spleen, lung, kidney, stomach, large intestine, small intestine and brain tissues. For the remaining rats, eyeball blood was taken 30 min after drug treatment, and their left rear ankle joints were taken to detect interleukin (IL)-1β and tumor necrosis factor (TNF)-α levels in serum by ELISA method; rutin, avicularin and quercitrin levels in the tissues of high-dose group were detected by HPLC; pharmacokinetic parameters were analyzed by using DAS 2.0. Our results showed that L. parasiticus decoction could significantly improve the paw edema situation of adjuvant arthritis model rats, and reduce IL-1β and TNF-α levels in rat serum. The in vivo efficacy substance analysis in rats showed that rutin was only present in the stomach with a small amount. AUC0-t of avicularin was stomach > small intestine > kidney, and the duration time in vivo was kidney=stomach > small intestine > lung > heart. AUC0-t of quercitrin was stomach > kidney > liver > heart > lung > spleen > small intestine > brain > large intestine > serum, and the duration time in vivo was kidney=liver=small intestine=brain=lung=spleen=heart=stomach > large intestine > serum. The research indicated that L. parasiticus decoction was effective in treating rats with adjuvant arthritis. Avicularin and quercitrin are important ingredients of L. parasiticus in antirheumatism therapy. The distribution of avicularin and quercitrin in rats were consistent with traditional understanding that L. parasiticus could attribute to the kidney and liver meridians.

OBJECTIVETo diagnose muscular dystrophy using Western blot (WB) by improving the method of the protein extraction.

METHODFirstly,we compared the effect of different sample buffer solutions and processing Methods on the extraction of muscle protein in rats,then selected the appropriate extracting method and the process of the muscular protein.

RESULTSWe put the selected sample buffer into the micro-sample,then mixed. The concentration of the extracting protein was much more,and the loss during the process was much less. We extracted enough protein in 62 cases. The protein bands were showed clearly by WB,and the abnormal protein bands were shown in some patients. Compared with the Results of immunohistochemical staining detected the severe abnormal expressions of Dys-R,Dys-C,and Dys-N in the specimens,we did not detect the corresponding target band in WB. We detected the target protein band of the specimens were abnormal position,light or normal staining in WB,while Dys were mildly expressed in immunohistochemical staining.

CONCLUSIONSThe improved protein extraction method can save the muscle tissue,and the protein bands can be used for diagnosing the muscular dystrophy. For clinically suspected patients with dystrophinopathy,if normal or mild deficiency is shown by immunohistochemistry,WB should be applied to detect the dystrophin protein band.

Animals ; Blotting, Western ; Dystrophin ; Humans ; Immunohistochemistry ; Muscular Dystrophies ; Protein Transport ; Rats ; Staining and Labeling

Objective To determine the audiological characteristics in 832 deaf children with biallelic causative mutations in GJB2 ,SLC26A4 gene .Methods The 832 patients received deafness gene screening ,553 were GJB2 gene biallelic causative mutations ,279 were SLC26A4 gene biallelic causative mutations .Patients were divided into four groups according to ages of hearing loss onset :<1 ,1~3 ,3~6 ,6~12 years old ,and the audiological character-istics and prevalence of GJB2 ,SLC26A4 gene mutations at different ages of onset .Results The prevalence of GJB2 gene mutations at four groups was 37 .97% (210/553) ,38 .34% (212/553) ,16 .27% (90/553) ,7 .41% (41/553) ,re-spectively ;the prevalence of SLC26A4 gene mutations at four groups was 25 .45% (71/279) ,44 .80% (125/279) , 20 .07% (56/279) ,9 .67% (27/279) ,respectively .The difference between GJB2 and SLC26A4 gene was significant(P=0 .001) .The prevalence of profound hearing loss with GJB2 gene mutations at four groups were 66 .67% (140/210) ,61 .32% (130/212) ,47 .78% (43/90) ,41 .46% (17/41) ,respectively .The difference was significant (P=0 .004) ,while the difference in 279 patients with SLC26A4 gene mutations was not statistically significant (P= 0 . 083) .Conclusion The age of hearing loss onset in patients with biallelic causative mutations in GJB 2 or SLC26A4 gene refers to 0~3 years -old ,hearing loss in patients with GJB2 ,SLC26A4 gene mutations gives priority to pro-found .The age of hearing loss onset is smaller ,the ratio of profound hearing loss is higher .Patients with severe and profound hearing impairment should be performed the genetic testing when the age of onset under 12 .

OBJECTIVETo explore a new method for finding more HIV/AIDS.

METHODSIn September 2009, newly reported HIV-infected individuals from May to August, 2009 in Dehong prefecture were asked to participate in a survey which requested demographic characteristics, history of high-risk behaviors and contact information of individuals with whom they had high risky contacts. People with risky contacts with HIV-infected cases (index cases) were also approached to participate in this survey and HIV testing was provided.

RESULTSA total of 342 HIV-infected individuals were newly reported and served as index cases from May to August, 2009. Among them, 47.1% (161/342) were transmitted by regular sexual partners. Through three-round surveys, 218 contacts were traced and among them, 84.9% (185/218) were traced by regular heterosexual partners. HIV positive rate of the tested was 34.7% (60/173) in the first-round tracing and 12.5% (1/8) in the second one. Among the 560 individuals (index cases and their high-risk contacts), the proportions of having regular heterosexual partners, non-regular and non-commercial sexual partners, commercial sexual heterosexual partners and men having sex men were 87.9% (492/560), 18.9% (106/560), 22.3% (125/560) and 0.3% (1/318), respectively, while the proportion of having never used condoms when having sex with the above four types sexual partners were 73.8% (363/492), 72.6% (77/106), 63.2% (79/125) and 0.0% (0/1), respectively.

CONCLUSIONAs an epidemiological method for HIV/AIDS finding, contact tracing identified a large number of HIV infectors who were traced by newly reported HIV-infected individuals.

Acquired Immunodeficiency Syndrome ; epidemiology ; Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Contact Tracing ; methods ; Female ; HIV Infections ; epidemiology ; Humans ; Infant ; Infant, Newborn ; Male ; Risk Factors ; Surveys and Questionnaires ; Young Adult

Objective To investigate current status of use of oral hypoglycemic drugs and insulin among adult patients with type 2 diabetes mellitus (T2DM) in urban community of Beijing.Methods In total,3297 T2DM patients aged more than 20 years from 15 urban communities of Beijing were studied.Their body weight,height,fasting plasma glucose level and glycosylated hemoglobin Alc (HbAlc) were measured.A door-to-door questionnaire survey on use of oral hypoglycemic drugs and insulin was conducted for them.All the T2DM patients surveyed were divided into four groups based on their received intervention.Results ①Of 3279 T2DM patients,454 (13.8％) received lifestyle intervention,971 (29.5％) used only one oral hypoglycemic drug,1179 (35.7％) with combined oral hypoglycemic drugs,and 693(21.0％) with insulin.②There was significant difference in average HbAlc among the four groups of T2DM patients with lifestyle intervention,only one oral hypoglycemic drug,combined oral hypoglycemic drugs,and insulin,with HbAI c of (7.0 ± 1.9) ％,(7.1 ± 1.5) ％,(7.4 ± 1.5 ) ％,and (7.5 ± 1.5 ) ％for them,respectively ( F =15.1,P ＜ 0.01 ).Proportions of the T2DM patients with HbAlc equal to or higher than 7.0％ were 32.2％,39.4％,52.1％ and 59.5％ for the four groups,respectively ( x2 =117.7,P ＜ 0.01 ).③In the T2DM patients with lifestyle intervention,32.2％ (146/454) of them with HbA1 c equal to or higher than 7.0％ were untreated with any oral hypoglycemic drug.In those with only one oral hypoglycemic drug,39.4％ (383/971) of them with HbAlc equal to or higher than 7.0％ were not treated with combined oral hypoglycemic drugs and/or insulin.In those with combined oral hypoglycemic drugs,52.1％ (614/1079) of them with HbAlc equal to or higher than 7.0％ were not received combined insulin treatment.④ Fasting plasma glucose level,treatment strategies,postprandial 2-h blood glucose level and length of the illness were independent risk factors for HbAlc level equal to or higher than 7.0％,with odds ratio (OR) of 1.757,1.256,1.175 and 1.031,respectively.⑤ In 2843 T2DM patients with oral hypoglycemie drugs and/or insulin treatment,1494 (52.6％ ) received biguanides and 693 received (24.4％ )insulin,respectively.Conclusions More than half of adult patients with T2DM do not meet the target of glycemic control of HbAlc less than 7.0％ in urban communities of Beijing,due to not active use of oral hypoglycemic drugs,and not timely adoption of combined use of oral hypoglycemic drugs and insulin therapy.