Aerosolized Ad5-nCoV booster vaccination elicited potent immune response against the SARS-CoV-2 Omicron variant after inactivated COVID-19 vaccine priming

The SARS-CoV-2 Omicron variant has become the dominant SARS-CoV-2 variant around the world and exhibits immune escape to current COVID-19 vaccines to some extent due to its numerous spike mutations. Here, we evaluated the immune responses to booster vaccination with intramuscular adenovirus-vectored vaccine (Ad5-nCoV), aerosolized Ad5-nCoV, a recombinant protein subunit vaccine (ZF2001) or homologous inactivated vaccine (CoronaVac) in those who received two doses of inactivated COVID-19 vaccines 6 months prior. We found that the Ad5-nCoV booster induced potent neutralizing activity against the wild-type virus and Omicron variant, while aerosolized Ad5-nCoV generated the greatest neutralizing antibody responses against the Omicron variant at day 28 after booster vaccination, at 14.1-fold that of CoronaVac, 5.6-fold that of ZF2001 and 2.0-fold that of intramuscular Ad5-nCoV. Similarly, the aerosolized Ad5-nCoV booster produced the greatest IFN{gamma} T-cell response at day 14 after booster vaccination. The IFN{gamma} T-cell response to aerosolized Ad5-nCoV was 12.8-fold for CoronaVac, 16.5-fold for ZF2001, and 5.0-fold for intramuscular Ad5-nCoV. Aerosolized Ad5-nCoV booster also produced the greatest spike-specific B cell response. Our findings suggest that inactivated vaccine recipients should consider adenovirus-vectored vaccine boosters in China and that aerosolized Ad5-nCoV may provide a more efficient alternative in response to the spread of the Omicron variant.

Study on spectrum-toxicity relationship of in vitro hepatotoxicity of aqueous extract from Euodia rutaecarpa / 中国药房

OBJECTIVE To study the spectru m-toxicity relationship of in vitro hepatotoxicity of aqueous extract from Euodia rutaecarpa. METHODS The aqueous extract from 16 batches of E. rutaecarpa from different habitats were prepared. The fingerprints of aqueous extract from E. rutaecarpa were established by ultra high performance liquid chromatography （UPLC） method and Similarity Evaluation System of TCM Fingerprint （2012A edition ），and common peaks were identified and the similarity was evaluated. Using normal human hepatocytes L 02 as subject ，inhibitory effect of aqueous extract from 16 batches of E. rutaecarpa to them were investigated. The spectrum-toxicity relationship of UPLC fingerprint of aqueous extract from E. rutaecarpa with the hepatotoxicity of hepatocytes L 02 was analyzed by grey relational analysis （GRA）and partial least squares regression analysis （PLSR）. The corresponding compound of the chromatographic peak with the greatest correlation with the in vitro hepatotoxicity of E. rutaecarpa were isolated ，prepared and identified. RESULTS There were 27 common peaks in UPLC fingerprints of aqueous extract from 16 batches of E. rutaecarpa ，with similarity of 0.375-0.995. Totally 9 peaks were confirmed ，i.e. neochlorogenic acid （peak 5），chlorogenic acid （peak 9），cryptochlorogenic acid （peak 10），caffeic acid （peak 12），rutin （peak 16），hyperin（peak 17），dehydroevotarine（peak 19），evotarine（peak 24），rutecarpine（peak 25）. The aqueous extract from 16 batches of E. rutaecarpa showed significant inhibitory effect on the growth of L 02 cells（P＜0.05 or P＜0.01），and the inhibitory rate ranged from 6.68％ to 67.95％. GRA showed that there were 18 common peaks with correlation degree greater than 0.8，which were peak 8＞peak 3＞peak 23＞peak 7＞peak 4＞peak 9＞peak 12＞peak 2＞peak 19＞peak 6＞ 4928381。E-mail：799247687@qq.com peak 15＞peak 5＞peak 1＞peak 17＞peak 21＞peak 26＞ peak 20＞peak 14 in descending order of correlation degree. PLSR showed that there were 14 peaks with regression coefficient＞0 and variable importance projection value ＞1，and the order of regression coefficient was peak 8＞peak 3＞peak 23＞ peak 2＞peak 7＞peak 4＞peak 12＞peak 9＞peak 19＞peak 5＞peak 17＞peak 26＞peak 10＞peak 15. Peak 8 had the greatest correlation with in vitro hepatotoxicity，and the corresponding compound of this peak was identified as 6-O-trans caffeoyl gluconic acid. CONCLUSIONS The in vitro hepatotoxicity of aqueous extract from E. rutaecarpa is the result of multiple component interaction，among which 6-O-trans caffeoyl gluconic acid shows closest relation with in vitro hepatotoxicity.

Study on protective effects of Shiyifang medicinal wine on knee osteoarthritis in rabbits based on TLR 4/MyD88/ NF-кB signaling pathway / 中国药房

OBJECTIVE To study the protective effect and mechanism of Shiyifang medicinal wine （SYF） on knee osteoarthritis（KOA）of rabbit induced by papain . METHODS Thirty-five rabbits were randomly divided into blank group ，model group，positive group （Diclofenac diethylamine emulsion 200 mg/kg），SYF high -dose group （386 mg/kg）and SYF low -dose group（97 mg/kg），with 7 rabbits in each group （all had 4 males and 3 females）. Except for the blank group ，the other groups ’ rabbits were injected 0.5 mL papain mixture （containing 2% papain and 0.03 mol/L L -cysteine）into the right knee cavity on day 1， 4 and 7，to replicate KOA model . Blank group was given constant volume of normal saline . From the 15th day ，drugs were applied to right hind knee joints of rabbits in each group ，twice a day for 20 days. At the same time ，the diameter of right knee joints of rabbits was measured by vernier calipers at day 0，8，14 and 35 to calculate swelling degree . After the experiment ，the levels of IL-1β，TNF-α and PGE 2 in synovial tissue were determined by enzyme -linked immunosorbent assay . Hematoxylin-eosin（HE） staining was used to prepare the sections of synovial tissue ，and the pathological changes were observed . The relative mRNA expressions of TLR 4，MyD88 and NF - кB p 65 in the TLR 4/MyD88/NF- кB signaling pathway were detected by real -time quantitative polymerase chain reaction . The relative protein expressions of TLR 4，MyD88，NF-кB p 65 and p -NF-кB p 65 were detect by Western blot . RESULTS Compared with blank group，the degree of knee swelling could be increased in model group ，the pathological damage of synovial tissue was more serious ，and the levels of IL -1β，TNF-α and PGE 2 were increased significantly in synovial tissue （P＜0.05）. The relative expression levels of TLR 4，MyD88，NF-кB p 65 mRNAs and TLR 4，MyD88，p-NF-кB p 65 proteins were significantly increased（P＜0.05）. Compared with model group ，swelling degree of right hind knee and the pathological injury degree of synovial tissue were significantly improved in each treatment group ，while the levels of IL -1β，TNF-α and PGE 2 in synovial tissue were significantly decreased （P＜0.05）. The relative mRNA expressions of TLR 4，MyD88 and NF -кB p 65 and relative protein expressions of TLR 4，MyD88（except for SYF low -dose group ）and p -NF-кB p 65 were all significantly decreased （P＜0.05）. CONCLUSIONS SYF shows protective effect on KOA induced by papain ，the mechanism of which is associated with decreasing the levels of IL -1β，TNF-α and PGE 2 and down -regulating TLR 4/MyD88/NF-кB signaling pathway .

Four-year changes in central fatness, risk of diabetes, and metabolic control in older adults: a cohort study with mediation analysis

Background/Aims@#Older adults are vulnerable to central obesity, while the association of changes in central fatness with risk of diabetes and metabolic control has not been investigated among this particular population. This study was aimed to address these issues. @*Methods@#A total of 1,815 adults aged ≥ 60 years without diabetes at baseline were followed for 4 years. Incident diabetes was ascertained based on plasma glucose, hemoglobin A1c, medical history, and/or the use of anti-diabetic drugs. Central fatness was assessed by waist circumference (WC), waist-height ratio (WHtR), and body roundness index (BRI). Logistic regression analyses were used to assess the association of changes in central fatness with risk of diabetes, along with dose-response and mediation analyses. @*Results@#During the 4-year follow-up, 177 participants developed diabetes. The risk of diabetes was increased by 42%, 41%, and 40% per 1 standard deviation increases in WC, WHtR, and BRI, respectively, in multivariable-adjusted models (all p < 0.01). Moreover, these relationships were all linearly-shaped (all pnonlinearity ≥ 0.11). Increases in WC, WHtR, and BRI correlated with increases in hemoglobin A1c, triglycerides-and-glucose index, triglycerides, white blood cell, and C-reactive protein (all p ≤ 0.04). Yet only changes in hemoglobin A1c and triglycerides-and-glucose index were identified as the possible mediators for risk of diabetes, with their mediating effect being about 35% and 21%, respectively. @*Conclusions@#Increases in central fatness were related to elevated risk of diabetes, and this association might be partly explained by the worsening of glycemic control and insulin resistance in older adults.

Study on regulatory mechanism of compatibility of ginseng and gecko dispensing granule on neuroendocrine immune network in model rats with kidney yang deficiency / 中国药房

OBJECTIVE To explore the regulatory mechanism of compatibility of ginseng and gecko dispensing granule on kidney yang deficiency model rats. METHODS Male SD rats were randomly divided into normal group （no modeling ，no administration），model group （modeling，no administration ），Jinkui shenqi pill group （modeling，dose of 2.33 g/kg），ginseng group（modeling，dose of 0.53 g/kg），gecko group （modeling，dose of 0.21 g/kg）and compatibility group （modeling，ginseng 0.53 g/kg and gecko 0.21 g/kg）. The body mass and anal temperature of rats were measured at different time points ；the serum levels of cAMP ，cGMP，CRH，ACTH，CORT，T，T3，T4，E2，IgG and IgM were measured ；the pathomorphological changes of adrenal gland ，thyroid gland and testis were observed ；mRNA expression of CRH ，thyroid stimulating hormone releasing hormone （TRH）and gonadotropin releasing hormone （GnRH）in hypothalamus were detected. RESULTS Compared with model group ，the anal temperature ，the levels of cAMP ，CRH，ACTH，CORT，T3，T and cAMP/cGMP ，T/E2 in serum and mRNA expressions of TRH and GnRH in hypothalamus were significantly increased in the compatibility group （P＜0.05 or P＜0.01）；the levels of cGMP，E2 and IgG in serum and mRNA expression of CRH in hypothalamus decreased significantly （P＜0.05 or P＜0.01）； the pathological injuries of adrenal gland ，thyroid gland and testis were all improved. Compared with ginseng or gecko dispensing granules alone ，the anal temperature and T/E 2 of rats in the compatibility group increased significantly ，and mRNA expression of CRH in hypothalamus decreasedsignificantly （P＜0.05 or P＜0.01）. CONCLUSIONS Thecompatibility of ginseng and gecko dispensing granule has a synergistic regulatory effect on kidney yang deficiency model rats ， the mechanism of which may be associated with hypothalamus-pituitary-adrenal axis ， hypothalamus-pituitary-thyroid axis ， hypothalamus-pituitary-gonad axis and neuroendocrine immune network formed by immune function. Compatible drugs are better than single drugs.

Cross-sectional Analysis of Emergency Department and Acute Care Utilization Among Medicare Beneficiaries.

BACKGROUND: We sought to develop a claims-based definition of unscheduled care to describe the use and role of the emergency department (ED) in providing unscheduled care to vulnerable older adult populations. METHODS: This study was a cross-sectional analysis of national 20% sample of Medicare beneficiaries included in the 2012 Chronic Condition Warehouse data set. We measured three outcomes: the number of ED visits per 1,000 Medicare beneficiaries, the proportion of all unscheduled ED and office-based visits occurring in the ED and the number of ED and non-ED unscheduled visits adjusting for risk factors. Each outcome was estimated for vulnerable subpopulations of Medicare beneficiaries with multiple chronic conditions (MCCs), dual eligibility, hospice enrollment, and skilled nursing facility use. RESULTS: A total of 10,717,786 Medicare beneficiaries were included with 33,696,461 potentially unscheduled care visits of which 5,192,235 (15%) occurred in the ED, 364,334 (1.1%) in facility-based urgent care, and 31,570,113 (84%) in ambulatory office settings. In regression analyses each subpopulation was more likely to visit the ED for unscheduled care services than the reference population of Medicare beneficiaries ages 65 to 80. Dual-eligible beneficiaries demonstrated higher ED visit rates and lower non-ED visit rates for unscheduled care. The subpopulation with MCCs uses both the ED and the non-ED setting for unscheduled care more so than any other group. CONCLUSIONS: Medicare beneficiaries, particularly vulnerable subpopulations, disproportionately visit the ED in comparison to physician offices for unscheduled care. Efforts to improve care coordination, measure quality, or reform payment to influence ED visitation should acknowledge these patterns and the unique availability of acute care services in the ED.

Efficacy and safety of Guhong injection for treating coronary microvascular disease: study protocol for a randomized controlled trial.

BACKGROUND: Coronary microvascular disease (CMVD) can be described as one of the cardiovascular diseases with normal coronary angiography but evidence of myocardial ischemia or microcirculatory lesions, often presenting as angina pectoris attacks. Coronary artery microtubular dysfunction is one of the pathogenic features of coronary heart disease, but its occurrence and development and the current CMVD-intervention therapy needs further research. Chinese traditional medicine (TCM) has advantages for the treatment of cardiovascular diseases. Hence, this article describes an ongoing randomized controlled clinical trial based on the theory of TCM for the purpose of evaluating the efficacy and safety of Guhong injection versus placebo in patients with CMVD. METHODS/DESIGN: This is a multicenter, randomized, parallel-arm, open-label, double-blind, placebo-controlled clinical trial. A total of 260 eligible patients will be allocated and randomly assigned, in a ratio of 1:1, to either the experimental group or the control group. The treatment course is 10 consecutive days, and with an 8-week follow-up. The primary outcome is therapeutic efficacy. Secondary outcomes include the quantitative score of TCM syndromes (a series of TCM symptoms and signs of coronary heart disease), the average frequency of anginal attacks, electrocardiogram (ECG) changes, inflammatory response, endothelial function indicators and myocardial metabolites. DISCUSSION: This trial is strictly designed in accordance with principles and regulations issued by the China Food and Drug Administration (CFDA). The results should provide high-quality evidence on the efficacy and safety of Guhong injection in the treatment of CMVD. TRIAL REGISTRATION: Chinese Clinical Trials Registry, ID: ChiCTR1900022902. Registered on 27 April 2019.

Comparative Study on the Effect of Raw Product and Different Processed Products of Gekko gecko on Kidney- yang Deficiency Model Mice Induced by Adenine / 中国药房

OBJECTIVE：To compare the effects of raw pro duct and different processed products of Gekko gecko on kidney-yang deficiency model mice induced by adenine. METHODS ：Totally 100 mice were randomly divided into blank group （n＝10）and modeling group （n＝90）. Modeling group was given adenine （50 mg/kg）intragastrically for 10 days to induce kidney-yang deficiency model ；blank group was given normal saline （0.2 mL/10 g）intragastrically. After modeling ，70 mice were randomly divided into model group ，positive group （Jinkui shenqi pill ，6.4 g/kg），G. gecko crude product group （1.2 g/kg）， wine-processed G. gecko group（1.2 g/kg）and oil-processed G. gecko group（1.2 g/kg）according to body weight and symptoms of kidney-yang deficiency ，with 14 mice in each group. Blank group and model group were given constant volume of normal saline intragastrically；administration groups were given relevant medicine intragastrically （0.2 mL/10 g），once a day ，for consecutive 14 d. During the experiment ，the symptoms and signs of mice in each group were observed. After last medication ，renal index ，testis index and serum levels of T ，CORT，BUN and Cr were measured ；HE staining method was used to observe the pathological changes of renal tissue of mice in each group. RESULTS ：Compared with blank group ，the mice in the model group suffered from performance of kidney-yang deficiency ，such as weight loss ，crouch and arch back ，chills and cold limbs ，and sparse body hair ， while renal index and serum levels of BUN and Cr were increased significantly （P＜0.01）. In renal tissue ，there were BA28117 pathological damages such as irregular arrangement of renal KY2016YB211tubular epithelial cells ，light staining of nucleus and edema of cytoplasm. Compared with model group ， performance of kidney-yang deficiency was improved to different extents in G. gecko crude product group and processed product groups（especially in wine-processed G. gecko group）；serum levelsof BUN and Cr were decreased significantly （P＜0.05 or P＜0.01）；pathological damage of renal tissue was alleviated in different degrees. In addition ，body weight of mice was increased significantly in G. gecko processed products groups （P＜0.01），and renal indexes of mice were decreased significantly in G. gecko crude product group and processed products groups （P＜0.05 or P＜0.01）. Compared with G. gecko crude product group ，renal index ，serum levels of BUN and Cr were significantly decreased in wine-processed G. gecko group（P＜0.01），and serum level of Cr was significantly decreased in oil-processed G. gecko group（P＜ 0.05）. CONCLUSIONS ：G. gecko crude product ，wine-processed G. gecko and oil-processed G. gecko all show a certain improvement effect on kidney-yang deficiency mice induced by adenine ，especially wine-processed G. gecko .

Do men with normal testosterone-oestradiol ratios benefit from letrozole for the treatment of male infertility?

RESEARCH QUESTION: Previous studies of aromatase inhibitors on male infertility have focused on men with low testosterone-oestradiol ratio of less than 10. Can aromatase inhibitors improve spermatogenesis in men with idiopathic male infertility with normal testosterone-oestradiol ratio? DESIGN: Prospective study of men with idiopathic severe oligozoospermia (sperm concentration <5 million/ml) carried out between February 2015 and March 2017. The objective was to assess if semen-analysis parameters improved after treatment with letrozole. Secondary objectives were to monitor the safety of letrozole in men, and to measure the alterations in serum FSH, LH, oestradiol and testosterone levels. RESULTS: Fifteen men with normal testosterone-oestradiol ratio (>10) were treated with letrozole 2.5 mg daily for 4 months. This produced a 5.5-fold increase in sperm concentration (P = 0.0068). All men had increased total serum testosterone and suppressed oestradiol levels after treatment, thus raising the overall testosterone-oestradiol ratio (P < 0.0001). Adverse effects from letrozole were relatively minor and included loss of libido (54%), headaches (25%), fatigue (21%), weakness (13%), loss of hair (8%) and dry mouth (8%). CONCLUSIONS: Letrozole improves sperm concentration and increases testosterone-oestradiol ratio for men with oligozoospermia who have normal testosterone-oestradiol ratio; its role in the treatment of male infertility may be extended to this group of patients. In addition, it is a relatively well-tolerated drug with no serious adverse effects.

Comparative study on neuropsychological tests for typical Alzheimer's disease, frontal variant of Alzheimer's disease and behavioral variant of frontotemporal dementia / 中华老年医学杂志

Objective To compare the neuropsychological features and other clinical and imaging characteristics among patients with typical Alzheimer's disease (TAD),frontal variant Alzheimer's disease (fvAD) and behavioral variant frontotemporal dementia (bvFTD).Methods Neuropsychological scales were used to evaluate the cognitive function and neuropsychiatric symptoms of nine patients with fvAD,30 patients with bvFTD and 32 patients with typical AD (TAD).The apolipoprotein E (ApoE) ε4 allele and brain imaging techniques,including the magnetic resonance imaging (MRI),11 C-Pittsburgh compound B positron emission tomography (PIB-PET),and 18F-deoxyglucose positron emission tomography (FDG-PET),were used for the differential diagnosis of fvAD and bvFTD.Results There were no significant differences in general cognitive function or daily activity in patients among the three groups.However,the cognitive subscale scores of Montreal Cognitive Assessment (MoCA) scale,including visual acuity and executive function,delayed recall,and orientation,were significantly different among the groups (Z =7.891,P =0.035;Z =7.412,P =0.031;Z=6.437,P=0.043,respectively).The clock drawing test (CDT) score of TAD group was higher than that in the bvFTD and fvAD groups (Z=8.673,P=0.020),while the neuropsychological questionnaire (NPI) scores of the bvFTD and fvAD groups were higher than those of the TAD group (Z=7.577,P =0.023).Meanwhile,the incidences of agitation,disinhibition and abnormal behavior of NPI in the bvFTD and fvAD groups were higher than those in the TAD group (x2 =11.139,P =0.004;x2 =6.481,P =0.039;x2 =6.812,P =0.033,respectively).The frequencies of the ApoE ε4 allele were 44.4％ in the fvAD group and 33.3％ in the bvFTD group,which were not significant different from that in the TAD group (40.6％).Furthermore,patients in the fvAD group were associated with decreased regional cerebral glucose metabolism and amyloid deposition in the temporalparietal cortex evaluated by FDG-PET and PIP-PET.Conclusions Neuropsychological evaluation is valuable for the differential diagnosis of TAD,fvAD and bvFTD.The frequencies of the ApoEε4 allele are similar among the three groups.FDG-PET and PIB-PET imaging plays an important role in the diagnosis of variant types of Alzheimer's disease.

Screening of full human anthrax lethal factor neutralizing antibody in transgenic mice / 生物工程学报

Anthrax is a highly lethal infectious disease caused by the spore-forming bacterium Bacillus anthracis. The major virulence factor of B. anthracis consists of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA binds with LF to form lethal toxin (LT), and PA binds with EF to form edema toxin (ET). Antibiotics is hard to work in advanced anthrax infections, because injuries and deaths of the infected are mainly caused by lethal toxin (LT). Thus, the therapeutic neutralizing antibody is the most effective treatment of anthrax. Currently most of the anthrax toxin antibodies are monoclonal antibodies (MAbs) for PA and US FDA has approved ABTHRAX humanized PA monoclonal antibody for the treatment of inhalational anthrax. Once B. anthracis was artificially reconstructed or PA had mutations within recognized neutralization epitopes, anti-PA MAbs would no longer be effective. Therefore, anti-LF MAbs is an important supplement for anthrax treatment. Most of the anti-LF antibodies are murine or chimeric antibodies. By contrast, fully human MAbs can avoid the high immunogenicity of murine antibodies. First, we used LF to immunize the transgenic mice and used fluorescent cell sorting to get antigen-specific memory B cells from transgenic mice spleen lymphocytes. By single cell PCR method, we quickly found two strains of anti-LF MAbs with binding activity, 1D7 and 2B9. Transiently transfected Expi 293F cells to obtain MAbs protein after purification. Both 1D7 and 2B9 efficiently neutralized LT in vitro, and had good synergistic effect when mixed with anti-PA MAbs. In summary, combining the advantages of transgenic mice, fluorescent cell sorting and single-cell PCR methods, this study shows new ideas and methods for the rapid screening of fully human monoclonal antibodies.

Pathogenic detection of infectious diarrhea in patients in a district of Beijing from 2011 to 2013 / 中国感染控制杂志

Objective To analyze the distribution and pulsed-field gel electrophoresis (PFGE)of pathogenic bacteria causing infectious diarrhea in a district of Beijing from 2011 to 2013,and provide basis for tracing infection sources.Methods A total of 1 179 stool specimens of infectious diarrhea from patients in a diarrhea outpatient department from January 2011 to December 2013 were collected,all isolated pathogens were identified by serotyping and PFGE analysis.Results 330 enteric pathogens were isolated from 1 179 specimens,the top 4 bacteria were Shi-gella spp .(28.18%,n=93),Salmonella spp .(20.91 %,n=69),Vibrio parahaemolyticus (13.33%,n =44),and diarrheagenic Escherichia coli (3.33%,n = 11 ).18 Shigella sonnei isolates were identified as 8 PFGE patterns, clustering similarity was close to 88%;69 Salmonella spp .strains belonged to 18 serotypes and 41 PFGE patterns, Salmonella senftenberg and Salmonella enteritidis had dominant patterns;no dominant PFGE patterns were obviously identified among 23 strains ofVibrio parahaemolyticus .Conclusion The serotypes and PFGE patterns of pathogenic bacteria in infectious diarrhea in past three years showed a wide distribution characteristics,the dominant PFGE patterns of Salmonella spp .and Shigella spp .need to be paid more attention,and outbreak of infectious diarrhea caused by Salmonella spp .and Shigella spp .should be alerted.

The imageology study of patients with the hippocampal sclerosis-associated medial temporal lobe epilep-sy / 中国神经精神疾病杂志

Objective To explore the incidence and imageological features of patients with the hippocampal sclerosis-associated medial temporal lobe epilepsy. Methods Seventy-eight patients with the medial temporal lobe epi?lepsy were recruited from our hospital during February 2012 to December 2013. Magnetic resonance imaging (MRI) and resonance spectroscopy (MRS) analysis were conducted in patients with with the hippocampal sclerosis-associated medial temporal lobe epilepsy, patients with epilepsy without the medial temporal lobe diseases and healthy controls. Results The incidence of hippocampal sclerosis was 58.97%among patients with medial temporal lobe epilepsy which were significantly higher compared with either healthy control group or patients with epilepsy without the medial tempo?ral lobe diseases. The average hippocampal volume of the medial temporal lobe epilepsy group(2305.68±814.61 mm3、2456.71±743. 60 mm3)was significantly smaller compared with either healthy controls or patients with epilepsy without the medial temporal lobe diseases. MRI revealed increased T2WI signal and hippocampal atrophy in 74.55%of patients with hippocampal sclerosis-associated medial temporal lobe epilepsy. Sclerosis was detected on the left side (52.17%) and bilateral hippocampus (19.57%). MRS showed that NAA/(Cr ± Cho) significantly reduced (0.58± 0.19) in the hip?pocampal sclerosis. Conclusions Hippocampal sclerosis may be the main imaging features of the medial temporal lobe epilepsy which are characterized by the hippocampal atrophy and high T2WI signal.

Production and immunogenicity analysis of conformation-stable fragment-C mutant of tetanus toxin / 生物工程学报

Tetanus is caused by tetanus toxin synthesized by Clostridium tetani. Fragment C (Hc), the 50 kDa carboxy-terminal portion of tetanus toxin, is nontoxic but has receptor protein binding activities, which has been evaluated as a potential new recombinant subunit vaccine to replace the traditional formaldehyde inactivated toxoid vaccine. It is easy for wild Hc (HcW) to form inter- and intra-molecular disulfide bonds and the different conformations changes unstably, which brings difficulties for vaccine production technology. In our study, the Cys 869 of HcW was mutated to A1a and the conformation-stable fragment-C mutant of tetanus toxin (HcM) was constructed. The HcM was expressed, fermented and purified and its stability, receptor binding and immunogenicity were evaluated. The result showed that the HcM got high-level expression and was purified to > 95% of purity. The purified HcM was conformation-stable at different temperature for different time and kept the binding activities with one of its receptor GT1b. Mice given three vaccinations by HcM developed a protective immune response and were 100% protected against an intraperitoneal administration of 1 x 10(3) 50% lethal doses (LD50s) of tetanus neurotoxin. All the results showed that the conformation-stable HcM had potent immunogenicity as a recombinant tetanus vaccine candidate with simple production process and similar immunogenicity with HcW. Whether for routine tetanus therapy or for countries to respond to unexpected events (war, earthquake or other disaster), it is of great significance.

Construction, expression, purification and characterization of mutant of Aspergillus flavus urate oxidase / 生物工程学报

We converted the TGC codon (307-309 bp) of Aspergillus flavus urate oxidase (UOX) gene to a GCC codon by using fusion PCR techniques to produce a C103A mutant. This gene was cloned into expression vector pET-42a (+) and then transformed into Escherichia coli BL21 (DE3). The mutant protein (UOX-Ala103) was expressed in soluble form at high levels after induction with IPTG The expressed rUOX-Ala103 accounted for about 45% of total bacterial proteins, rUOX-Ala103 of up to 98% purity was obtained after purified using hydrophobic interaction and anion exchange. Western blotting showed that the anti-UOX antibody specifically recognized rUOX-Ala103. The mutant protein showed a 60% increased in vitro biological activities compared with native protein, and performed a good activity of degrading the uric acid in vivo.

Expression and biological activity analysis of human-mouse chimeric antibody against anthrax protective antigen / 中华微生物学和免疫学杂志

Objective To express human-mouse chimeric antibody against anthrax protective anti-gen and to analyze its biological activities. Methods A new mammalian bipromoter expression vector was constructed with dihydrofolate reduetase(DHFR) gene as the selection and complication marker. First, the light and heavy chain variable region gene of the monoclonal antibody 5E1 were cloned by RT-PCR, at the same time the human IgG1 heavy chain constant region gene and kappa type constant region gene were cloned. Next, the human-mouse chimeric antibody genes were synthesized by fusion PCR. Then, the hu-man-mouse chimeric antibody gene were inserted into MCS of pSecTag and B1 to construct pSecTag-5E1L and B1-5E1H, respectively. Finally, heavy chain expression cassette excised from the B1-5E1H with Bgl Ⅱ/BamH Ⅰ was further cloned into the Bgl Ⅱ site of the pSecTag-5E1L to construct pSecTag-5E1. Plasmid pSecTag-5E1 was transfected into CHO(dhfr) engineering cells and high production cell clones that were screened by enhancing MTX concentration. After collecting medium and purifying chimeric antibody with af-finity chromatogram, purified chimeric antibody was analyzed by SDS-PAGE, Western blot. Results A sta-ble and high production cell line was acquired at MTX concentration 5×10~(-8) mol/L. Conclusion The hu-man-mouse chimeric antibodies were successfully expressed in CHO cells.

Study on the entrance of the anthrax toxin PA and LFn mediated EGFP into HeLa cells / 中华微生物学和免疫学杂志

Objective To study the role of protective antigen(PA)and N-terminal segment of lethal factor (LFn)in the entrance of EGFP(enhanced green fluorescent protein)into HeLa cells. Methods The DNA fragments encoding LFn and EGFP were amplified,respectively,and cloned into the plasmid pET-21 a(+)one after another to construct a recombinant plasmid pET-LFn-EGFP. The plasmid was txansformed into BL21 cells to express LFn-EGFP protein under the induction of IPTG. The protein was purified by Ni chelating chromatography. After incubation with LFn-EGFP in the presence of PA or not, the HeLa cells were analyzed by flow cytometry or laser confocal microscopy. Results The fusion protein LFn-EGFP was purified by over 90% homogeneity and retained the ability of LF to bind with PA when incubated with J774A.1 macrophage cells,and could get into HeLa cells. Conclusion The LFn-EGFP could enter the HeLa cells in a PA independent pathway. But PA could help more LFn-EGFP molecules enter into HeLa cells.

Research on repairing facial nerve injury of rabbits by neural stem cells and autologous fasia / 中国康复理论与实践

@#ObjectiveTo observe the effect on repairing facial nerve injury of rabbits by neural stem cells and autologous fasia. Methods22 rabbits with transected facial nerve were divided into 2 groups randomly, control group (8 rabbits,15 sides totally), which transected facial nerve were wrapped by autologous fasia, and treament group (14 rabbits, 20 sides totally), which were wrapped by neural stem cells and autologous fasia. Six weeks after transplantation, neuro-electrophysiological test, immunohistochemical examination were done. The number and thickness of myelin in the re-connected area of transected facial nerve were observed. ResultsThe transplanted animals recovered much better than that in control group (P<0.05). Immunohistochemical examination showed a great deal of BrdU positive cells around the re-connected area of transected facial nerve. Immunohistochemical staining also found plenty of regenerative myelins in this area in the treatment group. While in control group, there were no BrdU positive cells and only a few of regenerative myelins in the same area. ConclusionTransplantation of neural stem cells combined with autologous fasia might become the new method to treat facial nerve injury.

PREPARATION AND APPLICATION OF HUMAN PRP SYNTHYETIC PEPTIDE / 中国人兽共患病学报

Aim In order to improver the diagnostic rate of Prion disease, solve the lacking sources of nature PrP antigen. ,synthesize the human PrP peptide, prepare the antibody to PrP peptide further. Method A synthetic peptide with the sequence identical to the 15 residues of human PrP, as described by Kretzschmar[1], was synthesized by the solid-phase method. The synthetic peptide was coupled to bovine serum albumin(BSA)by the method of EDCI〔2〕. The polypeptide combined with BSA was used as antigen to immunize the rabbit and detected by immunomethod. Result The PrP polypeptide combined with BSA obtained immunogenicity and anti-PrP synthetic peptide antiserum was successfully obtained. Conclusion The preparation and application of human PrP synthetic peptide can substitute for nature PrP antigen partly. It has laid a foundation for further preparation of monoclonal antibody to PrP and the study of Prion disease.