Establishment of PCR assays and genetic polymorphism analysis of genes encoding Clostridium perfringens β2 toxin from different sources / 中华流行病学杂志

Objective: To establish and optimize PCR methods for the gene encoding of Clostridium perfringens β2 toxin (cpb2) and atypical-cpb2 (aty-cpb2), analyze the epidemiological characteristics and genetic polymorphism of the cpb2 of Clostridium perfringens in 9 Chinese areas from 2016 to 2021. Methods: The cpb2 of 188 Clostridium perfringens strains were examined by PCR; the cpb2 sequences were acquired by whole-genome sequencing to analyze the genetic polymorphism. Using Mega 11 and the Makeblastdb tool, a phylogenetic tree, and cpb2-library based on 110 strains carrying the cpb2 were produced. Using the Blastn technique, a comparison was made to discover sequence similarity between consensus-cpb2 (con-cpb2) and aty-cpb2. Results: The specificity of PCR assay for the cpb2 and aty-cpb2 was verified. The PCR results for cpb2 amplification were highly consistent with the whole-genome sequencing approach (Kappa=0.946, P<0.001). A total of 107 strains from nine regions in China carried cpb2, 94 types A strains carried aty-cpb2, 6 types A strains carried con-cpb2, and 7 types F strains carried aty-cpb2. The nucleotide sequence similarity between the two coding genes was 68.97%-70.97%, and the similarity between the same coding genes was 98.00%-100.00%. Conclusions: In this study, a specific PCR method for cpb2 toxin was developed, and the previous PCR method for detecting aty-cpb2 was improved. aty-cpb2 is the primary gene encoding of β2 toxin. There is a significant nucleotide sequence variance between the various cpb2 genotypes.

Study on the features of Clostridioides difficile infection among diarrhea patients in Kunming from 2018 to 2020 / 中华流行病学杂志

Objective: We analyze the characteristics of Clostridioides difficile (C. difficile) infection among diarrhea patients in Kunming from 2018 to 2020 and provide evidence for follow-up surveillance and prevention. Methods: A total of 388 fecal samples of diarrhea patients from four sentinel hospitals in Yunnan Province from 2018 to 2020 were collected. Real-time quantitative PCR was used to detect the fecal toxin genes of C. difficile. The positive fecal samples isolated the bacteria, and isolates were identified by mass spectrometry. The genomic DNA of the strains was extracted for multi-locus sequence typing (MLST). The fecal toxin, strain isolation, and clinical patient characteristics, including co-infection with other pathogens, were analyzed. Results: Among the 388 fecal samples, 47 samples with positive reference genes of C. difficile were positive, with a total positive rate of 12.11%. There were 4 (8.51%) non-toxigenic and 43 (91.49%) toxigenic ones. A total of 18 strains C. difficile were isolated from 47 positive specimens, and the isolation rate of positive specimens was 38.30%. Among them, 14 strains were positive for tcdA, tcdB, tcdC, tcdR, and tcdE. All 18 strains of C. difficile were negative for binary toxins. The MLST results showed 10 sequence types (ST), including 5 strains of ST37, accounting for 27.78%; 2 strains of ST129, ST3, ST54, and ST2, respectively; and 1 strain of ST35, ST532, ST48, ST27, and ST39, respectively. Fecal toxin gene positive (tcdB+) results were statistically associated with the patient's age group and with or without fever before the visit; positive isolates were only statistically associated with the patient's age group. In addition, some C. difficile patients have co-infection with other diarrhea-related viruses. Conclusions: The infection of C. difficile in diarrhea patients in Kunming is mostly toxigenic strains, and the high diversity of strains was identified using the MLST method. Therefore, the surveillance and prevention of C. difficile should be strengthened.

Phenotypic characteristics of wild-type plague phage growth in different experimental environments / 中国热带医学

@#Abstract: Objective To observe the phenotypic characteristics of 3 wild-type plague phages under different experimental environments, providing scientific evidence for the identification of phage biological characteristics and the study of their interaction with host bacteria in the future. Methods The sensitivity of 3 wild-type plague phages were detected by using liquid culture method, emisolid medium method and micro-liquid culture method based on OmniLog TM microbial identification system. Results The growth result based on LB liquid medium showed that the growth of plague phage 476 for 20-24 hours at both 28 ℃ and 37 ℃was better than that of plague phages 087 and 072204 at 37 ℃, and the growth of plague phages 087 was better than that of plague phages 072204 at 37 ℃. With the attenuated plague bacterium EV76 as the host bacterium, phage 476 was able to form visible plaque on double-layer agar medium for 20-20 hours at both 28 ℃ and 37 ℃, phages 087 and 072204 were only able to form opaque plaque on double-layer agar medium for 20-24 hours at 37 ℃. The growth results based on OmniLogTM system showed that when plague phage was lysed in EV76 strain at 33 ℃, the first row appeared as a straight line with a peak of no more than 100 in the 96-well microplate curve chart. As the phage quantity decreased, the dilution plate appeared with growth curve similar to EV76 strain in turn, and the color of tetrazolium dyes in the experimental wells gradually deepened as the phage number decreased and the host bacteria number increased. Therefore, it indicates that phage 476 was sensitively at both 28 ℃ and 37 ℃, while phage 087 and 072204 were temperature-dependent only at 37 ℃ to attenuated plague bacterium EV76. Conclusions The lysing ability of 3 wild-type plague phages are temperature-dependent, and the growth results are consistent under the three experimental conditions.

Effects of different pretreatment agents on primary tooth dentin bonding durability / 口腔疾病防治

Objective@#To explore the effects of different pretreatment agents on primary tooth dentin bonding durability.@*Methods @#Forty-two retained primary molars were selected, 24 of which were cut along the mesial and distal directions; thus, 48 samples were obtained for shear bond strength tests, and the other 18 teeth were used for nanoleakage tests. According to different pretreatments, both experimental samples were divided randomly into three groups (Group A: distilled water pretreatment group; Group B: 2% chlorhexidine pretreatment group; Group C: 10 mg/mL resveratrol pretreatment group). The test specimens were prepared, the shear bond strength was tested, and interfacial nanoleakage evaluation and scanning electron microscope observation were performed to evaluate the effects of different pretreatment agents on the bonding interface immediately and after aging for one hour with 10% sodium hypochlorite aqueous solution.@*Results @#The immediate shear bond strength results showed that there was no significant difference among the three test groups. After aging, the shear bond strength of Group C was significantly higher than that of Group A and Group B (P<0.05). After aging, the shear bond strength of Group A was significantly lower than the immediate shear bond strength (P<0.05), whereas there was no significant difference in shear bond strength before and after aging in Group B and Group C (P>0.05). For Group C, there was no significant difference in interfacial nanoleakage before and after aging. In addition, among the three groups, Group C had the lowest interfacial nanoleakage (P<0.05).@*Conclusion@# Both chlorhexidine and resveratrol pretreatment can improve the adhesion durability of deciduous dentin, but the effects of resveratrol are better than those of chlorhexidine.

The correlation between the timed up and go test and fall risks in elderly frail patients / 中华老年医学杂志

Objective:To evaluate the correlation between the timed up and go(TUG)test and fall risks in elderly frail patients.Methods:From July to September 2019, elderly frail patients who were treated at the cardiovascular department of our hospital were enrolled.Basic clinical data and fall-related information of patients were collected.Patients were divided into the fall group and the non-fall group.Results on the body mass index(BMI), TUG, 4-meter maximum walking speed(4 m MWS)and Barthel index were compared between the two groups.The correlation between TUG and each indicator was examined.Multivariate Logistic regression analysis was used to analyze the correlation between the TUG and falls in elderly patients.Results:A total of 96 eligible patients were enrolled, including 35 in the fall group and 61 in the non-fall group.The average TUG time was longer in the fall group than in the non-fall group(16.45±6.44 s vs.10.17±2.91 s, t=-6.556, P<0.001). The correlation analysis results showed that the TUG was correlated with falls and 4 m MWS( r=0.582 and 0.875, both P<0.001). Multivariate Logistic regression analysis showed that the TUG( OR=1.201, 95% CI: 1.111-1.470, P=0.004)and 4 m MWS( OR=1.146, 95% CI: 1.063-1.244, P=0.015)were risk factors for falls. Conclusions:The TUG is correlated with fall risks in elderly frail patients and should be recommended as a routine test in clinical practice.

Comparison of clinical efficacy of two different pin arrangements for external fixation of femoral neck fracture / 中国骨伤

OBJECTIVE@#To retrospectively analyze the clinical efficacy of external fixation in the treatment of femoral neck fracture with two different pin layout.@*METHODS@#From April 2000 to April 2018, 140 cases of femoral neck fracture were treated with closed reduction and percutaneous pin external fixation, among them 121 cases were followed up for more than 1 year, including 31 cases in traditional group, 12 males and 19 females, aged 45 to 74(65.4±8.4) years;90 cases in modified group, 39 males and 51 females, aged 12 to 75 (64.5±7.8) years. In traditional group, the first needle was put on the femoral talus, the second and third needles were put under the tension line, and the three needles were not on the same line in the lateral phase; in modified group, the first needle was drilled into the lateralcortex of the femur, obliquely penetrating the distal and proximal end of the femoral talus fracture, and the other two needles were drilled into the medial cortex of the femoral neck and the femoral talus, respectively. The operation time, hospital stay, postoperative ambulation time, femoral neck shortening rate, fracture healing time, fracture healing rate and femoral head necrosis rate of the two groups were observed and compared. Harris hip function score was used one year after operation.@*RESULTS@#These 121 patients were followed-up, the follow up time of traditional group was 13 to 45(30.5±11.4) months;the follow-up time of modified group was 14 to 120(34.5±12.5) months. There was no significant difference in operation time, hospital stay and femoral head necrosis rate between two groups (@*CONCLUSION@#Compared with the traditional group, the modified group has the advantages of lower femoral neck shortening rate, shorter fracture healing time, higher fracture healing rate and higher Harris hip function score.

Effect of dentin surface treatments on the bond strength of resin-modified glass ionomer cement / 口腔疾病防治

@#Resin-modified glass ionomer cement (RMGIC) has good physical, chemical and biological properties and is suitable for the treatment of deciduous caries, aged root surface caries and wedge-shaped defects. Surface treatment is a common method to improve bonding strength, which can improve physical and chemical retention between different components. This paper mainly introduces the current research status of the influence of different dentin surface treatment methods on the bonding strength of RMGIC. At present, the common dentin surface treatment methods are pretreatment, acid etching, laser treatment, etc. The pretreatment agent can improve the bond strength of RMGIC by increasing the surface area and porosity of dentin. The bond strength of RMGIC could be effectively improved after the dentin was treated by an acid-etching bonding system. The dentin was treated with a laser to obtain a higher bonding strength. However, whether the use of resin adhesives will affect the release of fluoride ions in RMGIC into the deep dentin and thus affect the repair effect also needs further research.

Treatment of elderly patients with aneurysmal subarachnoid hemorrhage: comparison between surgical clipping and endovascular coil embolization / 国际脑血管病杂志

Objective To compare the treatment effect of surgical clipping and endovascular coil embolization for aneurysmal subarachnoid hemorrhage (aSAH) in the elderly.Methods Elderly patients with aSAH (aged ＞65 years) treated in Shanghai Pudong Hospital from January 2009 to December 2017 were enrolled retrospectively.They were divided into craniotomy clipping group and endovascular intervention group according to the treatment strategy.The Glasgow Outcome Scale w as used for short-term outcome assessment at discharge,4-5 were defined as good outcome,and 1-3 were defined as poor outcome.Longterm follow-up w as performed to assess clinical outcomes using the modified Rankin Scale,0-2 w as defined as good outcome and 3-6 w ere defined as poor outcome.The clinical and imaging information,perioperative complications,short-term and long-term clinical outcomes,and long-term imaging outcomes w ere compared between the two groups.Multivariate logistic regression analysis was used to assess the independent influencing factors of clinical outcomes.Results A total of 68 elderly patients with aSAH were enrolled.Of these,47 (69.1％) received endovascular coil embolization and 21 (30.9％) underwent microsurgical clipping.There were no significant differences between the two groups in the incidence of perioperative complications,short-term and long-term adverse outcomes,and imaging recurrence rate.Multivariate logistic analysis showed that smoking (odds ratio [OR] 36.319,95％ confidence interval [CI] 3.530-373.640;P =0.003),modified Fisher grade (OR 20.406,95％ CI 2.022-205.964;P =0.011) and World Federation of Neurological Societies (WFNS) grade (OR 4.686,95％ CI 1.012-21.692;P=0.048) were the independent risk factors for short-term poor outcomes in elderly patients with aSAH.Conclusion Both endovascular intervention and microsurgical clipping are safe and effective treatments for elderly patients with aSAH.

Treatment of elderly patients with aneurysmal subarachnoid hemorrhage: comparison between surgical clipping and endovascular coil embolization / 国际脑血管病杂志

Objective@#To compare the treatment effect of surgical clipping and endovascular coil embolization for aneurysmal subarachnoid hemorrhage (aSAH) in the elderly.@*Methods@#Elderly patients with aSAH (aged >65 years) treated in Shanghai Pudong Hospital from January 2009 to December 2017 were enrolled retrospectively. They were divided into craniotomy clipping group and endovascular intervention group according to the treatment strategy. The Glasgow Outcome Scale was used for short-term outcome assessment at discharge, 4-5 were defined as good outcome, and 1-3 were defined as poor outcome. Long-term follow-up was performed to assess clinical outcomes using the modified Rankin Scale, 0-2 was defined as good outcome and 3-6 were defined as poor outcome. The clinical and imaging information, perioperative complications, short-term and long-term clinical outcomes, and long-term imaging outcomes were compared between the two groups. Multivariate logistic regression analysis was used to assess the independent influencing factors of clinical outcomes.@*Results@#A total of 68 elderly patients with aSAH were enrolled. Of these, 47 (69.1%) received endovascular coil embolization and 21 (30.9%) underwent microsurgical clipping. There were no significant differences between the two groups in the incidence of perioperative complications, short-term and long-term adverse outcomes, and imaging recurrence rate. Multivariate logistic analysis showed that smoking (odds ratio [OR] 36.319, 95% confidence interval [CI] 3.530-373.640; P=0.003), modified Fisher grade (OR 20.406, 95% CI 2.022-205.964; P=0.011) and World Federation of Neurological Societies (WFNS) grade (OR 4.686, 95% CI 1.012-21.692; P=0.048) were the independent risk factors for short-term poor outcomes in elderly patients with aSAH.@*Conclusion@#Both endovascular intervention and microsurgical clipping are safe and effective treatments for elderly patients with aSAH.

A New High-throughput Real-time PCR Assay for the Screening of Multiple Antimicrobial Resistance Genes in Broiler Fecal Samples from China / 生物医学与环境科学（英文）

OBJECTIVE@#Antimicrobial resistance (AMR) has become a global concern and is especially severe in China. To effectively and reliably provide AMR data, we developed a new high-throughput real-time PCR assay based on microfluidic dynamic technology, and screened multiple AMR genes in broiler fecal samples.@*METHODS@#A high-throughput real-time PCR system with an new designed integrated fluidic circuit assay were performed AMR gene detection. A total of 273 broiler fecal samples collected from two geographically separated farms were screened AMR genes.@*RESULTS@#The new assay with limits of detection ranging from 40.9 to 8,000 copies/reaction. The sensitivity rate, specificity rate, positive predictive value, negative predictive value and correct indices were 99.30%, 98.08%, 95.31%, 99.79%, and 0.9755, respectively. Utilizing this assay, we demonstrate that AMR genes are widely spread, with positive detection rates ranging from 0 to 97.07% in 273 broiler fecal samples. blaCTX-M, blaTEM, mcr-1, fexA, cfr, optrA, and intI1 showed over 80% prevalence. The dissemination of AMR genes was distinct between the two farms.@*CONCLUSION@#We successfully established a new high-throughput real-time PCR assay applicable to AMR gene surveillance from fecal samples. The widespread existence of AMR genes detected in broiler farms highlights the current and severe problem of AMR.

Polymorphism analysis of virulence-related genes among Candida tropicalis isolates / 中华医学杂志(英文版)

BACKGROUND@#Adhesion, biofilm formation, yeast-hyphal transition, secretion of enzymes, and hemolytic activity are all considered important factors in Candida tropicalis infection. However, DNA sequence data for this pathogen are limited. In this study, the polymorphism and heterogeneity of genes agglutinin-like sequences (ALS)2, Lipase (LIP)1, LIP4, and secretory aspartyl proteinase tropicalis (SAPT)1-4 as well as the relationship between phenotype and genotype were analyzed.@*METHODS@#This study started in August 2013, and ended in July 2017. The complete length of ALS2, LIP1, LIP4, and SAPT1-4 of 68 clinical C. tropicalis isolates was sequenced. Single nucleotide polymorphisms (SNPs) as well as insertions and deletions (indels) were identified within these genes. In addition, phenotypic characteristics of the virulent factors, including adhesion and the secretion of aspartyl proteinases and phospholipases, were determined.@*RESULTS@#There were 73, 24, 17, 16, 13, and 180 SNPs in the genes LIP1, LIP4, SAPT1, SAPT2, SAPT3, and SAPT4, respectively. Furthermore, 209 SNPs were identified in total for the gene ALS2. Interestingly, large fragment deletions and insertions were also found in ALS2. Isolate FXCT 01 obtained from blood had deletions on all 4 sites and showed the lowest adhesion ability on the polymethylpentene surface. In addition, isolates with deletions in the regions 1697 to 1925 and 2073 to 2272 bp displayed relatively low abilities for adhesion and biofilm formation, and this phenotype correlated with the deletions found in ALS2. LIP1, SAPT4, and ALS2 displayed great heterogeneity among the isolates. Large deletions found in gene ALS2 appeared to be associated with the low ability of adhesion and biofilm formation of C. tropicalis.@*CONCLUSION@#This study might be useful for deeper explorations of gene function and studying the virulent mechanisms of C. tropicalis.

Effect of resvertrol on body mass and biochemical indexes of diabetic cataract model rats / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To study the effect of resveratrol on body mass and biochemical parameters of diabetic cataract model rats.<p>METHODS: Thirty rats were divided into blank group, model group and treatment group with 10 rats for each. Rats in the blank group were not treated. Rats in the model group and the treatment group were modeled. Rats in the treatment group were intervened with astragalol. Lens opacity, body mass, body growth and biochemical indexes of rats in each group were detected. The right eyeballs of rats in the three groups were stained with HE.<p>RESULTS: The rats in the blank group had transparent lens and no opacity; the degree of lens opacity and the overall score of cataract in the treatment group were better than those in the model group(<i>P</i><0.05); at 3rd and 5th weeks, the body weight and length of the rats in the treatment group were higher than those in the model group, and the levels of FPG and 2hPG were lower than those in the model group(<i>P</i><0.05); after 5wk, the biochemical indexes in the treatment group were significantly improved compared with those in the model group(<i>P</i><0.05). <p>CONCLUSION: Astragall can improve the degree of lens opacity in diabetic cataract rats, alleviate the effect of disease on the body weight of rats, regulate the level of blood sugar and biochemical indicators in rats, and alleviate the progress of cataract.

Discussion on strategy of development of forestry schistosomiasis control programs of China in new period / 中国血吸虫病防治杂志

Based on the literature research, this paper states the mechanism and measures of the Oncomelania snail control by the schistosomiasis prevention forests, reviews the development process of forestry schistosomiasis control programs, analyzes the new situation and problems, and then puts forward some countermeasures and suggestions about the national forestry program for schistosomiasis control in order to provide new ideas for the goal of schistosomiasis elimination in China in 2025 and the construction of national forestry program for schistosomiasis control.

Analysis of proficiency testing results for pathogenic bacteria in laboratory animal in six years / 中国比较医学杂志

Objective To analyze the result of proficiency testing(PT)of detection activities for Laboratory animal pathogenic bacteria in 2011 and 2013-2017. To further improve the detection capacity of laboratory animal testing agency,and promote PT to be carried out in future. Methods During the six years(2011 and 2013 -2017), the National Institutes for Food and Drug Control conducted a total of six(seven projects)PT activities of laboratory animal pathogen bacteria. We analyzed the overall trend and the exposed problems by summarizing the result data of the PT in 6 years. Results A total of 45 laboratories in the country including 20 provinces and cities participated in the PT. The PT projects included Mycoplasma pulmonis, Clostridium piliformis, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella spp.,Klebsiella pneumoniae and Bordetella bronchiseptica. The satisfaction rates were 75%,87.5%,80.0%, 78.6%,93.3,96.2% and 88.0%, respectively. The main reasons of unsatisfactory results were for lack of incubation time,select errors of suspicious bacteria, biochemical identification errors, report writing errors and not timely feedback results. Conclusions The level of domestic laboratory animal pathogenic bacteria detection is gradually increased to achieve the desired goal through continuous proficiency testing activities.

Research progress in long non-coding RNAs involved in invasion and metastasis of breast cancer / 基础医学与临床

Long non-coding RNAs(lncRNAs) defined as RNAs of more than 200 nucleotides in length don't en-code protein but are closely related to the development of breast cancer.Plenty of lncRNAs are dysregulated in breast cancer,and they can regulate the invasion and metastasis of breast cancer through various mechanisms such as transcription and post-transcriptional regulation which is greatly important to guide the treatment and prognosis of breast cancer.

Genotypic analysis of Mycobacterium tuberculosis strains collected from He'nan Province by 24-locus variable-number tandem-repeat typing and spoligotyping / 中华传染病杂志

Objective To investigate the genotype of M.tuberculosis in He'nan Province.Methods A total of 668 M.tuberculosis clinical strains collected in difference regions of He'nan Province during 2015 were genotyped by two standard methods,including classical 24-locus mycobacterium interspersed repetitive unit variable-number tandem-repeat (MIRU-VNTR) typing and spoligotyping.Results The 668 isolates were divided into 11 clusters and 35 patterns by spoligotyping.Among the 558 Beijing strains,546 were typical Beijing strains and the other 12 were atypical Beijing strains.Among the 110 non-Beijing strains,eight were new strains and the remaining 102 non-Beijing strains were divided into 10 families.There were 76 isolates belonging to T family,including 59 of T1 families,7 of T2 families,and 10 of T3 families.The 668 strains were divided into 550 gene patterns by standard 24-locus VNTR,including 508 un-clustered patterns and 160 clustered into 42 clusters.The largest cluster contained 21 strains,the other clusters contained 2-20 strains.Conclusion Beijing strain is still the most prevalent M.tuberculosis in He'nan Province.

Stromal cell-derived factor-1 inducing metastasis squamous cell carcinoma of head and neck induced via integrin ανβ3-CXC chemokine receptor 4/7 biological axis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the influence of LM609/AMD3100/CCX754 on chemotactic capability, cytoskeleton, and expression of integrin ανβ3 protein of squamous cell carcinoma of head and neck (SCCHN) cell line PCI-13 induced by stromal cell-derived factor-1 (SDF-1) in vitro.</p><p><b>METHODS</b>Migration assays, flow cytometry and immunofluorescence were used to observe the effects of SDF-1, LM609, AMD3100 and CCX754 on the migration, cytoskeleton and the expression of integrin ανβ3 protein in PCI-13 cell lines.</p><p><b>RESULTS</b>SDF-1 favored PCI-13 cell migration, pseudopod formation, and activities of integrin ανβ3 phosphorylation. LM609, AMD3100, and CCX754 blocked all these effects.</p><p><b>CONCLUSIONS</b>SDF-1 can induce metastatic SCCHN by integrin ανβ3-CXC chemokine receptor (CXCR) 4/CXCR7 axi. LM609, AMD3100, and CCX754 and can reduce the regulation of SDF-1 on SCCHN activity.</p>

Functional Investigation of a GRIN2A Variant Associated with Rolandic Epilepsy / 神经科学通报·英文版

N-methyl-D-aspartate receptors (NMDARs), a subtype of glutamate-gated ion channels, play a central role in epileptogenesis. Recent studies have identified an increasing number of GRIN2A (a gene encoding the NMDAR GluN2A subunit) mutations in patients with epilepsy. Phenotypes of GRIN2A mutations include epilepsy-aphasia disorders and other epileptic encephalopathies, which pose challenges in clinical treatment. Here we identified a heterozygous GRIN2A mutation (c.1341T>A, p.N447K) from a boy with Rolandic epilepsy by whole-exome sequencing. The patient became seizure-free with a combination of valproate and lamotrigine. Functional investigation was carried out using recombinant NMDARs containing a GluN2A-N447K mutant that is located in the ligand-binding domain of the GluN2A subunit. Whole-cell current recordings in HEK 293T cells revealed that the N447K mutation increased the NMDAR current density by ~1.2-fold, enhanced the glutamate potency by 2-fold, and reduced the sensitivity to Mg inhibition. These results indicated that N447K is a gain-of-function mutation. Interestingly, alternative substitutions by alanine and glutamic acid at the same residue (N447A and N447E) did not change NMDAR function, suggesting a residual dependence of this mutation in altering NMDAR function. Taken together, this study identified human GluN2A N447K as a novel mutation associated with epilepsy and validated its functional consequences in vitro. Identification of this mutation is also helpful for advancing our understanding of the role of NMDARs in epilepsy and provides new insights for precision therapeutics in epilepsy.

Structure optimum and biological activity studies of APC/Asef peptide inhibitors / 上海交通大学学报（医学版）

Objective: To design and synthesize APC/Asef peptide inhibitors, and investigate the relationship between the structures and affinity of peptides. Methods: Based on crystal structure of the APC-MAI-150 complex, on the one hand, 3-phenylpropane, Glu-Glu (GG) and β-Ala-Ala (β-AA) were used to replace carbobenzoxy (CBZ) at the N terminal of MAI-150 to bind -NH2; on the other hand, -CN, -NO2, -NH2, and -F were replaced the parahydroxyl on the sixth tyrosine (Tyr) side chain benzene at the N terminal of MAI-150. And seven peptides were synthesized. Fluorescence polarization was applied to test peptide affinity, and molecular design laboratory-3 (MDL-3), MDL-4 and MDL-5 were docked into APC protein based on the results of activity. The structure-activity relationship of the three peptides was studied by combining the binding patterns of computer docked peptide and APC protein. Results: Among the seven peptides, the N terminal 3-phenylpropane linked -NH2 of peptide MDL-5 had the highest affinity, which IC50 was 2.35 μmol/L. Compared with MDL-5, the affinity of MDL-6 and MDL-7 were significantly reduced. The para-hydroxyl on the sixth Tyr side chain benzene at the N terminal replaced with -NH2 (MDL-3) and -F (MDL-4), which the affinity were higher than -CN (MDL-1) and -NO2 (MDL-2). However, the affinity of newly synthesized peptides was lower than MAI-150. Conclusion: Transforming peptide N terminal linked -NH2 and the para-hydroxyl group on the sixth Tyr side chain benzene doesn't help to improve the affinity of peptides.

Progress in Application of Measuring Skeleton by CT in Forensic Anthropology Research / 法医学杂志

Individual identification by measuring the human skeleton is an important research in the field of forensic anthropology. Computed tomography （CT） technology can provide high-resolution image of skeleton. Skeleton image can be reformed by software in the post-processing workstation. Different skeleton measurement indexes of anthropology, such as diameter, angle, area and volume, can be measured on section and reformative images. Measurement process is barely affected by human factors. This paper reviews the literatures at home and abroad about the application of measuring skeleton by CT in forensic anthropology research for individual identification in four aspects, including sex determination, height infer, facial soft tissue thickness measurement and age estimation. The major technology and the application of CT in forensic anthropology research are compared and discussed, respectively.