Establishment of a screening cell model for cannabinoid type 1 receptor agonists / 药学学报

Cannabinoid receptors are one of the most expressed G protein-coupled receptors in the central nervous system, which are potential drug targets for inflammation, pain and drug abuse. Cannabinoid receptors are composed of type 1 receptor (CB1R), type 2 receptor (CB2R) and other receptors, of which CB1R plays a vital role in regulating central memory, cognition, and motor function. Therefore, screening CB1R agonists has potential value in treating nervous system diseases. In this study, the intracellular loop 3 (ICL3) domain of CB1R was replaced with a circular-permutated enhanced green fluorescent protein (cpEGFP). After infecting HEK 293T cells with lentivirus particles, we obtained a stable cell line that was overexpressed human CB1R-cpEGFP after puromycin selection. The interaction between receptor agonists and CB1R led to the change of receptor conformation, resulting in de-protonation of the EGFP, and enhancing the fluorescence intensity. Therefore, active CB1R compounds could be verified by measuring the fluorescence intensity. Using CB1R agonist arachidonyl-2′-chloroethylamide (ACEA) as a positive control to evaluate the reliability of this model, studies have shown that ACEA could induce receptor activation and increase fluorescence intensity, while antagonist rimonabant inhibited receptor activation with unchanged fluorescence intensity. In conclusion, this study successfully constructed a fluorescent probe screening model for CB1R agonists.

Clinical efficacy and safety of tumor-specific individualized multi-target DC-CIK in the treatment of advanced non-small cell lung cancer / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：评价肿瘤特异性个体化多靶点树突状细胞-细胞因子诱导的杀伤细胞（DC-CIK）治疗晚期非小细胞肺癌（NSCLC）患者的临床疗效和安全性。方法：回顾性分析2019年10月1日至2022年10月31日东部战区总医院生物治疗科行肿瘤特异性个体化多靶点DC-CIK治疗晚期NSCLC患者的临床资料。统计NSCLC患者的临床疗效和不良反应，分析治疗前后血清中肿瘤标志物的变化，FCM检测患者治疗前后的淋巴细胞亚群和各种细胞因子的表达情况，用质谱仪检测治疗前后靶点的变化。结果: 共入组52例晚期NSCLC患者，其中女性21例、男性31例；年龄32~71岁，平均年龄（50.97±10.72）岁，中位年龄47.5岁。经DC-CIK治疗后，CR 0例，PR 0例，SD 27例，PD 25例。与治疗前比较，DC-CIK治疗后：（1）CEA和CYFRA21-1水平无显著改变，CA125水平显著低于治疗前（P<0.01）；（2）治疗后患者淋巴细胞亚群无显著变化；（3）治疗后患者外周血IL-2、IL-4、IFN-γ和TNF-α水平显著升高（均P<0.01），IL-6、IL-10及IL-17水平无明显变化；（4）治疗后靶点数下降明显。DC-CIK治疗过程中无严重不良反应发生。结论: 晚期NSCLC患者行肿瘤特异性个体化多靶点自体DC-CIK治疗是安全的，能使患者产生抗肿瘤免疫反应并得到一定的临床获益。

The screening model for dopamine receptor agonists by a dopamine sensor / 药学学报

Parkinson's disease (PD) is a degenerative disease of the central nervous system due to the loss or death of dopaminergic neurons in the substantia nigra. Clinically, levodopa is the most effective and commonly used drug for PD treatment. However, long-term levodopa therapy is prone to motor complications and other side effects caused by excessive peripheral dopamine production, which has become an urgent problem to be solved in PD treatment. Dopamine receptor (DR) agonists are similar to dopamine. They can directly stimulate postsynaptic dopamine receptors, produce the same effect as dopamine, delay the application of levodopa as much as possible, and reduce complications caused by long-term use of levodopa. Therefore, screening effective dopamine receptor agonists has become a key issue in the study and treatment of PD. In order to establish a rapid, stable and reliable method for dopamine receptor agonist screening, this study used the human dopamine receptor 2 (DRD2) gene fused with a circular permuted EGFP (cpEGFP) to construct a recombinant gene, packaged with lentiviral vector, and the vector replaced the parted inner transmembrane domain of the third intracellular loop (ICL3) of genetically-encoded GPCR-activation based (GRAB) sensors. The fluorescence of GPCR-fused cpEGFP is regulated by conformational changes mediated by the interaction of dopamine receptor agonists with GPCRs without altering GPCR activity. The HEK293T cells were infected with viral vector, screened by puromycin to select highly expressed cells. Dopamine receptor agonists (including dopamine, bromocriptine mesylate, cabergoline, pramipexole) were used as positive drugs to explore the best screening and detection conditions, establishing a stable model to evaluate the dopamine receptor agonist. The results showed that the optimal filter for the dopamine receptor agonist in this study was the cell seeding count of 7×104, and the effective concentration of the positive drug was 1-100 µmol·L-1. In addition, pretreated with 10 µmol·L-1 dopamine receptor antagonists (including chlorprothixol hydrochloride, domperidone, and sulpiride), the positive fluorescence signal of overexpressed DRD2-cpEGFP HEK293T cells could not be detected when exposed to 10 µmol·L-1 dopamine receptor agonists, which proved that dopamine receptor antagonists could block the activity of dopamine receptor agonists, so they cannot activate dopamine receptor allosteric, indicating that the model has good specificity and can also be used for the screening and detection of new dopamine receptor antagonists. In summary, the study constructs a stable dopamine sensor detection system, which can effectively screen potential dopamine receptor agonists. The operation procedures are simple and rapid. And it can be used for a large-scale screening providing a fundamental methodology for drug development and PD treatment targeted on DRD2.

Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica / 口腔疾病防治

Objective@# To investigate the transcriptomic changes in primary human oral keratinocytes (pHOKs) after coculture with Prevotella melaninogenica (P.m) and to verify the changes in human oral keratinocyte (HOK) cell lines.@*Methods@# pHOK was isolated and cocultured with P.m for 0, 4 and 24 h. Total RNA was extracted, a gene library was constructed, transcriptional sequencing was performed, differentially expressed genes (DEGs) were analyzed, gene ontology (GO) pathway analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed, and the validation of DEGs was performed by qRT-PCR and Western Blot in the HOK and P.m coculture cell model. @*Results @#1 788 DEGs were detected between the 4 h group and control group, including upregulated DEGs such as lymphocyte cytosolic protein 1(LCP1), keratin 7 (KRT7) and Cilia and flagella associated protein 251(CFAP251) and downregulated DEGs such as FERM, ARH/RhoGEF and Pleckstrin domain protein 1 (FARP1), WW domain containing transcription regulator 1(WWTR1) and Discoidin, CUB and LCCL domain-containing protein 2 (DCBLD2). 1 832 DEGs were detected between the 24 h group and control group, including upregulated DEGs such as LCP1, complement C1s(C1S), kynureninase (KYNU) and downregulated DEGs such as phosphoserine aminotransferase 1 (PSAT1), FARP1 and FKBP prolyl isomerase 10 (FKBP10). There were 1 090 common differentially expressed genes (cDEGs) in the 4 h and 24 h groups, including LCP1, KYNU and long intergenic nonprotein coding RNA 958 (LINC00958). The GO pathways were mainly enriched in response to lipopolysaccharide and the molecules of bacterial origin and apical part of the cell. KEGG pathway analysis revealed enrichment in the interleukin-17 (IL-17) signaling pathway, tumor necrosis factor (TNF) signaling pathway, Toll-like receptor (TLR) pathway, etc. We verified the expression of a cDEG, Myosin1B (MYO1B), and qRT-PCR and Western Blot analysis showed that MYO1B expression was significantly upregulated between the control group and the P.m cocultured group (P<0.001), and its expression followed a time-dependent and concentration-dependent manner. @*Conclusion@#P.m played an important role in the transcriptome of oral keratinocytes.

Tracing investigation and analysis of a Clostridium botulinum food poisoning incident in Bayingolin Mongolian Autonomous Prefecture, Xinjiang / 中华预防医学杂志

To analyze a suspected case of Clostridium botulinum food poisoning in Bayingolin Mongolian Autonomous Prefecture, Xinjiang and to help validating the diagnosis and providing technical support for clinical treatment. The basic information and clinical manifestations of food poisoning cases were investigated by using the epidemiological method of food safety accidents. The botulinum toxin genes in the samples were detected by real-time PCR and inoculation of KM mouse. The enriched bacteria were further purified and validated. PFGE and cluster analysis were performed on five isolates. Clostridium botulinum type A was detected in two homemade fermented bean samples and stool lavage fluid samples of three patients from enriched samples by toxin test and real-time PCR, and were further validated after isolation of Clostridium botulinum. PFGE showed 100% homology among five isolates. Five isolates of bacteria isolated from the stool lavage fluid of three patients and two homemade fermented bean curd were identified as the same source through PFGE. The cause of this food poisoning cases is food pollution of Clostridium botulinum type A.

Mechanism of Dabuyuan Jian in Promoting Neurogenesis of Hippocampus in APP/PS1 Double Transgenic Dementia Mice Based on Wnt/β-catenin Signaling Pathway / 中国实验方剂学杂志

Objective::To investigate the possible mechanism of Dabuyuan Jian to promote neurogenesis by studying the effect of Dabuyuan Jian on Wnt/β-catenin signaling pathway in hippocampus of amyloid precursor protein/presenilil(APP/PS1) mice. Method::Totally 30 5-month-old APP/PS1 mice and 10 wild mice were divided into control group, model group, donepezil group (6.5×10-4 g·kg-1·d-1) and Dabuyuan Jian group (13.2 g·kg-1·d-1), and given drugs by gavage for 30 days. The control group and the model group were given an equal volume of saline. The learning and memory of mice were evaluated by water maze. The pathological changes of hippocampal neurons were observed by hematoxylin-eosin (HE) staining. The immunohistochemistry (IHC) was used to detect label positive cells of 5-bromodeoxyuridine (Brdu), adrenocortical hormone (Dcx) and neuronal nuclear antigen (NeuN). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western bolt were used to detect the mRNA and protein expression levels of β-catenin and glycogen synthase kinase-3β(GSK-3β) in hippocampus of mice. Result::Compared with the control group, the latency and swimming total distance of the model group were significantly increased (P<0.01), while the number of crossing platforms and the target quadrant time were significantly reduced (P<0.01). Compared with the model group, the platform latency and the total swimming distance of the donepezil group and the Dabuyuan Jian group were decreased (P<0.05, P<0.01), while the number of crossing platforms and the target quadrant time were increased (P<0.05). Compared with the control group, the number of neurons in the hippocampal dentate gyrus (DG) area of the model group was decreased, and necrotic neurons were observed. Compared with the model group, the number of neurons in the hippocampal DG area of the mice in the donepezil group and the Dabuyuan Jian group was increased, while the number of necrotic neurons was decreased. Compared with the control group, the number of positive cells labeled with Brdu, Dcx and NeuN in the hippocampal DG area of the model group was significantly decreased (P<0.01). Compared with the model group, the number of positive cells labeled with Brdu, Dcx and NeuN in the hippocampal DG area of the donepezil group and the Dabuyuan Jian group was increased (P<0.05, P<0.01). Compared with the control group, gene and protein expression levels of β-catenin in the hippocampus of the model group were significantly decreased (P<0.01), whereas gene and protein expression levels of GSK-3β were significantly increased (P<0.01). Compared with the model group, gene and protein expression levels of β-catenin in hippocampus of donepezil group and Dabuyuan Jian group were increased (P<0.05, P<0.01), while gene and protein expression levels of GSK-3β were increased (P<0.05, P<0.01). Conclusion::Dabuyuan Jian could promote hippocampal neurogenesis in APP/PS1 double transgenic mice by regulating Wnt/β-catenin signaling pathway.

Dabuyuan Jian Improves Hippocampal Synaptic Plasticity of APP/PS1 Double Transgenic Mice by Regulating BDNF/TrkB/CREB Signal Pathway / 中国实验方剂学杂志

Objective:To observe the effect of Dabuyuan Jian on the synaptic plasticity of hippocampus and the brain derived neurotrophic factor （BDNF）/tyrosine kinase receptor （TrkB）/cyclic adenosine phosphate reactive element binding protein （CREB） signaling pathway in amyloid precursor protein/presenilil （APP/PS1） mice， and to explore its possible mechanism for improving synaptic plasticity. Method:Totally 36 APP/PS1 mice were divided into model group， donepezil group（6.5×10-4 g·kg-1·d-1） and Dabuyuan Jian group （13.2 g·kg-1·d-1）， and another wild mice were set as control group. The mice in control group and model group received an equal volume of saline， and the mice in each group received drugs by gavage for 30 days. The learning ability and memory of mice in each group were detected by Morris water maze. The pathological changes of neurons and synapses in the hippocampus of each group were observed by Nissl staining and Golgi staining. The expression levels of postsynaptic density protein 95 （PSD95） and synaptophysin （SYN） in hippocampus of each group were detected by immunofluorescence （IF）. The protein expression levels of BDNF， TrkB， CREB and phosphorylated CREB （p-CREB） in hippocampus were detected by Western blot. Result:As compared with the control group， the platform latency and total swimming distance of the model group were increased in the model group （P<0.01）， with decreased times of crossing the platform and staying time in the target quadrant （P<0.01）， the intracellular Nissl bodies of neurons in hippocampal CA3 area decreased or disappeared in model group， with decreased number of neurons and dendritic branches and decreased density of dendritic spine in hippocampal CA3 area of the mice （P<0.01）， and the protein expression levels of SYN， PSD95， BDNF， TrkB and p-CREB in hippocampus of mice were also decreased in model group （P<0.05， P<0.01）. As compared with the model group， the platform latency and total swimming distance were decreased in the donepezil group and Dabuyuan Jian group （P<0.05， P<0.01）， with increased times of crossing platform and staying time in target quadrant （P<0.05， P<0.01）， the number of Nissl bodies of neurons in hippocampal CA3 area was increased in the donepezil group and Dabuyuan Jian group， with increased number of neurons and dendritic branches and increased density of dendritic spine in hippocampal CA3 area of the mice， and the protein expression levels of SYN， PSD95， BDNF， TrkB and p-CREB in hippocampus of mice were increased in the donepezil group and Dabuyuan Jian group （P<0.05， P<0.01）. Conclusion:Dabuyuan Jian can improve the synaptic plasticity of APP/PS1 double transgenic mice， and its mechanism may be related to its up-regulation of BDNF/TrkB/CREB signal pathway in mouse hippocampus.

Anxiety and depression in middle school and college students with acne / 中国学校卫生

Objective@#To investigate anxiety and depression among young acne patients, to provide a reference for the comprehensive treatment and to improve the quality of life of the patients.@*Methods@#Totally 157 students with acne of a major skin disease (acne group) and 157 healthy subjects (control group) selected from 244 and 297 students with and without acne were evaluated with the Zung’s Depression Self-rating Scale (SDS) and the Zung’s Anxiety Self-rating Scale(SAS). The SDS and SAS scores at different levels of sex and disease state were compared between two groups.@*Results@#The positive detection rate of depression symptoms and anxiety symptoms in acne group was 59.20% and 69.40%,respectively, and the combined detection rate of anxiety and depression symptoms was 52.90%; the detection rate of depression symptoms and anxiety symptoms in control gruop was 32.50% and 25.50%,respectively, with anxiety and depression symptoms accounted for 20.40%, the differences were statistically significant(χ2=22.63, 60.81, 35.69, P<0.01). Compared with acne group, there were higher SDS and SAS scores in acne group(Z=5.95, 9.16, P<0.01). In the acne group, the SDS and SAS scores of males were higher than females. There were significant differences in the SDS and SAS scores of the samples with different incidence(H=55.67, 43.83, P<0.01).@*Conclusion@#Students with acne are more likely to suffer from depression and anxiety. Compared with healthy population, moderate and severe acne patients have more serious depression and anxiety symptoms. It is suggested that the medical staff in school should take appropriate psychological intervention and treatment to facilitate the comprehensive rehabilitation of the young students with acne.

Ultrastructure Characteristics of Different Chinese Medicine Syndromes of Helicobacter pylori-Correlated Gastric Diseases / 中国结合医学杂志

OBJECTIVE@#To explore the ultrastructure characteristics of patients with dampness-heat of Pi (Spleen)-Wei (Stomach) syndrome (DHPW) and Pi-qi deficiency syndrome (PQD), both of which are Helicobacter pylori (Hp)-correlated gastric diseases (HPCG), and implicate a helpful hint for the clinical microcosmic syndrome differentiation.@*METHODS@#Fourteen gastric mucosa samples from 6 chronic gastritis (CG) and 6 active peptic ulcer (including 8 DHPW, 4 PQD) as well as 2 healthy volunteers were collected and tested for Hp infection. The ultrastructure of gastric mucosa was observed under the transmission electron microscope (TEM).@*RESULTS@#Among 14 gastric mucosa samples, 8 of them were Hp positive (6 DHPW and 2 PQD), which were all accordance with the results screened by supermicro-pathological method. Under TEM, the normal gastric mucosa, with tidy microvilli and abundant in mucus granules, mitochondria and rough endoplasmic reticulum distributed evenly, and with smooth nucleus membrane. But in those specimens of DHPW with Hp infection, microvilli were presented with burr shape. Especially, those samples from dampness-heat syndrome with predominant heat type (DHSH) patients were more obvious, with microvilli damaged, mitochondria concentrated and distributed in disorder, secretory tubule extended. In dampness-heat syndrome with predominant dampness type (DHSD) patients, mucus granules aggregated obviously, mitochondria swelled and blurred, and rough endoplasmic reticulum crowded. For 2 samples of DHPW without Hp infection, their microvilli were intact, with mitochondria increased and gathered but well-distributed, and secretory tubule extended mildly. In 2 PQD patients with Hp positive, the specimens of microvilli were sparse, and their mucus granules and mitochondria were decreased, with fractured crests and vacuole, secretory tubules extension to nucleus membrane, and rough endoplasmic reticulum extension in a pool-like way, and nucleus condensed. The 2 samples from PQD patients without Hp infection were characterized with intact microvilli, decreased mitochondria, fractured crest and extended rough endoplasmic reticulum in a pool-like way.@*CONCLUSION@#It's obviously different in ultrastructure of DHPW and PQD patients under TEM, which may give a helpful hint for the microcosmic syndrome differentiation of HPCG.

Effect of zoladronate on the proliferation and osteogenic differentiation of rat bone mesenchymal stem cells / 华西口腔医学杂志

OBJECTIVE@#To evaluate the effect of zoledronate acid (ZA) on the proliferation and osteogenic differentiation of rat mesenchymal stem cells (BMSCs).@*METHODS@#The BMSCs isolated from the SD rats were cultured with different concentrations of ZA (1, 5, 10, and 20 μmol·L), and the contro1 group received the same volume of culture medium but without ZA. Cell counting kit-8 was used to detect proliferation activity in each group. Alkaline phosphatase (ALP) staining and alizarin red staining were used to detect the osteogenic differentiation ability in each group. The gene expression levels of ALP, bone morphogenetic protein-2 (BMP-2), typeⅠcollagenase (COL-Ⅰ), runt-related transcription factor-2 (Runx-2), zinc finger structure transcription factor (Osx), osteocalcin (OCN), and osteopontin (OPN) were evaluated by real-time quantitative polymerase chain reaction (qRT-PCR).@*RESULTS@#Zoledronate at 1 μmol·L⁻¹ concentration had no effect on the proliferation and osteogenic differentiation of BMSCs. No significant difference was observed between this group and the control group (P>0.05). When the ZA concentration was more than 1 μmol·L⁻¹, ZA inhibited the proliferation and osteogenic differentiation of BMSCs, and the effect was concentration dependent. The difference between each group and the control group was statistically significant (P<0.05). At ZA concentration of 5 μmol·L⁻¹, ZA enhanced the expression of ALP, BMP-2, COL-Ⅰ, Runx-2, Osx, OCN, and OPN (P<0.05). However, at ZA concentration of more than 5 μmol·L⁻¹, the expression levels of osteogenicrelated genes in each group was lower than those of the control group (P<0.05).@*CONCLUSIONS@#Low ZA concentration has no effect on the proliferation and osteogenic differentiation of BMSCs. ZA at 5 μmol·L⁻¹ concentration inhibits the proliferation but promotes the osteogenic differentiation of BMSCs. High ZA concentration inhibits the proliferation and osteogenic differentiation of BMSCs.

Control of imported mosquito-borne diseases under the Belt and Road Ini-tiative / 中国血吸虫病防治杂志

Mosquito is a vector of many infectious diseases,and it is recognized a leading killer of human in the world.After the Belt and Road Initiative launches,more are countries involved and the international communication and cooperation are sig-nificantly growing in China.Therefore,the risk of imported infectious diseases is increasing as well,some mosquito-borne dis-eases which have been well controlled or seldom seen in China,will be more risky to cause locally transmission from imported cases and become the threat to people's health in China.This paper reviews the risk of major imported mosquito borne-diseases to China,and discusses the control strategy as well,so as to provide the suggestion for entry-exit inspection and control of im-ported mosquito-borne diseases in China.

Clinical observation of Conbercept intravitreal injection combined with retinal laser photocoagulation therapy on macular edema secondary to retinal vein occlusion / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To study the efficacy of conbercept intravitreal injection combined with retinal laser photocoagulation therapy and simple laser photocoagulation therapy on macular edema (ME) secondary to retinal vein occlusion (RVO).METHODS:Forty-eight patients (53 eyes) with macular edema secondary to retinal vein occlusion diagnosed by clinical examination from October 2014 to March 2015 were retrospectively analyzed.Among them,28 patients (31 eyes) were treated with conbercept intravitreal injection combined with retinal laser photocoagulation,which was defined as Group A.And simple laser group contained 20 patients (22 eyes),which was defined as Group B.The clinical data including the patients' best-corrected visual acuity (BCVA) and central retinal thickness (CMT) before treatment and 1wk and 3mo after treatment were observed.RESULTS:Followed up for 3mo,the average BCVA values of A and B were 0.44±0.25,0.56±0.24,respectively and the average CMT were 330.50 ± 121.71,354.67 ± 102.79μm at first week of treatment.There was no significant difference in BCVA and CMT of Group A compared with Group B.There was statistically significant in BCVA and CMT of Group A and Group B compared with before treatment (P＜0.05).The average BCVA values of A and B were 0.24±0.18,0.39±0.20,respectively and the average CMT were 252.62 ± 83.01,332.67 ± 102.33μ m at third month of treatment.There were statistically significant differences between the two groups and compared with before treatment (P＜0.05),and Group A was superior to Group B.CONCLUSION:Conbercept intravitreal injection combined with retinal laser photocoagulation therapy and simple laser photocoagulation treatment of macular edema secondary to retinal vein occlusion are both effective that macular edema is significantly reduced,and vision is stable and improved.But for serious cases,conbercept intravitreal injection can reduce retinal edema at first,then combine with retinal laser photocoagulation which has obvious therapeutic effect and it is better than simple laser photocoagulation treatment.

Comparative studies on cariogenic bacteria of the root surface before and after post-core crown restoration in aged people / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the changes in the quantity of colonizing Streptococcus mutans(S. mutans) and Actinomyces on the root surface plaque before and after post-core crown restoration of the mandibular first molars in the elderly patients.</p><p><b>METHODS</b>A total of 30 elderly patients, each with one post-core crown restoration of the mandibular first molar, were randomly chosen to participate in the studies. Patients with mandibular first molars with post-core crown restoration and those with healthy contralateral mandibular first molars were divided into the test and control groups, respectively. Root surface plaques of the two groups were collected before tooth preparation, 72 h after preparation, one week after preparation, and one month after restoration. S. mutans, Actinomyces naeslundii (A. naeslundii) and Actinomyces viscosus (A. viscosus), were identified using colony morphology, biochemical techniques, and polymerase chain reaction (PCR). Plaque count was measured using microbial colony count.</p><p><b>RESULTS</b>The number of S. mutans and A. viscosus and A. naeslundii in the test group, which was statistically significant (P<0.05), increased 72 h after preparation. The quantities of S. mutans, A. viscosus, and A. naeslundii one week after preparation were significantly different (P<0.05). The plaque count of S. mutans, A. viscosus, and A. naeslundii in the test group decreased one month after restoration (P<0.05).</p><p><b>CONCLUSION</b>The quantities of S. mutans, A. viscosus and A. naeslundii increase one week after preparation but decrease one month after restoration. The finding suggests that dentists should educate patients about plaque control during the early period after tooth preparation.</p>

Advances in recently identified coronaviruses / 病毒学报

Coronaviruses are a large family of viruses which include viruses that cause the common cold and severe acute respiratory syndrome (SARS) in humans and other diseases in animals. There are considerable genetic diversities within coronaviruses due to their wide rang hosts and their special gene replication and transcription mechanisms. During this process, gene recombinations often occur, resulting in novel subtype or coronavirus emerge constantly. Of note are SARS-like-CoVs and novel HCoV-EMC identified in 2012. This minireview summarized major advances of recently identified coronaviruses, focusing on the genome structures and interspecies jumping mechanism of coronavirus.

Mifepristone repairs alteration of learning and memory abilities in rat model of depression / 药学学报

This study is to investigate the amelioration effect of glucocorticoid receptor (GR) antagonist mifepristone on the changes of learning and memory abilities in rat model of depression. In the present study, a 35-day rat chronic unpredictable stress (CUS) model was used to observe both depression-like behaviors with sucrose preference test and open-field test and learning and memory-associated behaviors with Morris water maze test. A total of 45 male adult Sprague-Dawley rats were randomly assigned to three groups of equal size: control group (CON); CUS group (CUS); CUS + mifepristone group (CM). Animals in CM group were first exposed to CUS for 14 days, and then were administered with 50 mg x kg(-1) x d(-1) of mifepristone with continued CUS procedure. Corticosterone EIA Kit was used to detect the concentration of plasma corticosterone (CORT). Nissl staining was used to observe the structure of hippocampus. The results demonstrated that CUS exposure induced both depressive-like and learning and memory-associated behaviors and these deficits were reversed by mifepristone. Compared to CON group, the concentration of plasma CORT increased significantly in CUS group. CUS exposure damaged the structure of hippocampus, whereas mifepristone had an amelioration effect. Together, the structural deficits of hippocampus resulting from long-term stress exposure, which could contribute to the impairment of learning and memory in depression, are reversed by the GR receptor antagonist mifepristone.

Analysis of EDA gene mutation for a family affected with X-linked hypohidrotic ectodermal dysplasia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect potential mutations of EDA gene for a Chinese family affected with X-linked hypohidrotic ectodermal dysplasia (XLHED).</p><p><b>METHODS</b>Genomic DNA was extracted from peripheral blood of the proband, his relatives and 50 non-related healthy controls. Exonic sequences of the EDA gene were subjected to polymerase chain reaction amplification and direct sequencing.</p><p><b>RESULTS</b>A c.467G> A mutation (R156H) was detected in exon 3 of the EDA gene in the proband, his mother, 2 uncles, and 1 aunt. The same mutation was not detected in the 50 non-related healthy controls.</p><p><b>CONCLUSION</b>A c.467G>A mutation of the EDA gene probably underlies the disease in the family.</p>

Molecular detection assays for 2012 identified novel human coronavirus (HCoV) and probe modification with locked nucleic acid (LNA) / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To develop and optimize the molecular detection assays for recently identified human coronavirus (HCoV) infection.</p><p><b>METHODS</b>Based on the 208 base pair(bp) sequence of novel HCoV reported by HPA of UK, we designed and obtained several pairs of primer (F-1, R-1; F-2, R-2) and Taqman probes (TZ1,TZ2) for detection of novel HCoV. Two of probes were modified with LNA (LNA-TZ1, LNA-TZ2). Then, RT-PCR and various real time RT-PCR assays were developed and optimized in this study. We also compared our assays with the real time RT-PCR assays reported recently by Europe team based on upE or ORF1b target.</p><p><b>RESULTS</b>The RT-PCR or real time RT-PCR assays for novel HCoV were developed without cross-reactivity with other HCoV and several common respiratory viruses using clinical specimen panel. The analytical sensitivity of assays were less than 50-500 copies per reaction and the detection was improved when Taqman probe modified with LNA-tagged, compared to no LNA-tagged in real time RT-PCR assays. The upE and LNA-TZ1 based assays were better than others.</p><p><b>CONCLUSION</b>The molecular detection sensitivity and specificity of TaqMan-based real time PCR assay could be improved when probe tagged with LNA. The upE or LNA-TZ1 based real time RT-PCR assay was recommend for detection of novel HCoV. This study laid a foundation for improving the performance of novel HCoV detection.</p>

Genetic diversity in cultivated populations of Dendrobium officinale / 中草药

Objective: To investigate the genetic diversity in cultivated populations of Dendrobium officinale, providing the basis for the protection of germplasm resources and breeding of new varieties. Methods: A molecular marker RAPD was used to analyze the genetic diversity of 14 populations of D. officinale and a population of D. devoninum from Yiwu, Tiantai, Jiande, Wuyi in Zhejiang Provinces and Malipo, Menghai in Yunnan Provinces. Results: Fifteen primers screened from 225 RAPD primers were used in the PCR amplification of 105 samples of D. officinale and 3 samples of D. devoninum; And 138 bands were generated among which 133 bands were polymorphic. The percentage of polymorphic bands (PPB) was 96.38%; The percentage of polymorphic loci (PPL) for 14 populations of D. officinale ranged from 10.79% to 76.26% was 45.32% (mean). Conclusion: Cultivated D. officinale of main producing places in whole country is rich in genetic diversity. Genetic relationship of D. officinale has a obvious correlation with its geography provenance, but nothing to do with the cultivation place.

Study on 11 metal correction of mental element contents in Dendrobium officinale / 中国中药杂志

<p><b>OBJECTIVE</b>To find out the variation of 11 mental element contents in Dendrobium officinale with different germplasms and harvesting ages, the results can provide scientific basis for the quality evaluation and the breeding of D. officinale.</p><p><b>METHOD</b>32 samples with 1-3 ages were collected from cultivated fields of Zhejiang and 11 samples were collected from markets. The 11 mental element contents of samples were determined by ICP-MS or AAS.</p><p><b>RESULT</b>The average contents of K, Ca, Mg, Mn, Zn, Cr, and Cu were 1,205.23, 766.82, 158.25, 31.06, 4.28, 8.28, and 0.97 mg x kg(-1), the contents of As, Hg, Pb, and Cd were all in limits except Cd content of one sample exceeded the standard limit 0.07 mg x kg(-1); germplasms and physiological ages impacted mental elements contents accumulation significantly.</p><p><b>CONCLUSION</b>There were rich essential mental elements in D. officinale. D. officinale from Zhejiang province and medical materials from market were all safe; the breeding of D. officinale can increase the contents of essential mental elements and reduce contents of heavy mental elements; the effect of physiological age on metal elements contents was related to each element's physiological and biochemical function.</p>

Variation of monosacchride composition of polysacchrides in Dendrobium officinale by pre-column derivatization HPLC method / 中国中药杂志

The monosacchride composition of polysacchrides in Dendrobium officinal of different germplasms, physiological ages and closely related species were determined by pre-column derivatization HPLC. The results showed that the absolute and relative volumes of all monosacchrides were significantly different between D. officinale and its closely related species, different germplasms and physiological ages of D. officinale. Absolute peak areas of mannose ranged from 0.854 x 10(7) to 10.340 x 10(7) in closely related species of D. officinale, ranged from 1.467 x 10(7) to 8.475 x 10(7) in different germplasms of D. officinale and were 4.411 x 10(7) (2.577 x 10(7)-6.516 x 10(7)), 5.528 x 10(7) (3.179 x 10(7)-8.475 x 10(7)) and 3.601 x 10(7) (1.467 x 10(7)-5.888 x 10(7)), respectively, in one to three physiological ages of D. officinale. The ratio of mannose to glucose peak areas (relative peak area) ranged from 0.976 to 16.599 in closely related species of D. officinale and from 2.679 to 7.831 in different germplasms of D. officinale. Only the relative peak areas of D. pendulum and D. primulinum were in the range of different germplasms of D. officinale in all tested samples. The results revealed the variation of monosacchride composition of polysacchrides in D. officinale. Monosacchride composition of D. officinale could be altered by breeding new varieties and controlling harvesting season. Most adulterants of D. officinale could be ruled out according to the relative peak areas of D. officinale, providing a basis for quality control and resources training of D. officinale.