Construction of serial deletants and chimeras of multi-kringle containing molecules and primary analysis of their functions

In comparison of amino acid sequences of 4 kringles of both macrophage stimulating protein (MSP) and hepatocyte growth factor (HGF), consensus motif sequence was determined. According to this consensus sequence, a pair of universal primers were designed. In combination with specific upstream or downstream primer of MSP or HGF respectively, serial fragments containing variant number of kringle (from 1 to 4) can be obtained by once PCR. By ligating the C terminal and N terminal fragments with different combination, serial deletants and chimeras of MSP and HGF were constructed. Sequence analysis showed that the degeneracy for universal primers and the sequences of those constructed deletants and chimeras are desired. Biological assay of these deletants revealed that wild type MSP can inhibit the growth of some tumor cell lines and that kringle 1 of MSP is essential for function as that of HGF.

Melatonin improves synapse development by PI3K/Akt signaling in a mouse model of autism spectrum disorder

Autism spectrum disorders are a group of neurodevelopmental disorders involving more than 1100 genes,including Ctnnd2 as a candidate gene.Ctnnd2knockout mice,serving as an animal model of autis m,have been demonstrated to exhibit decreased density of dendritic spines.The role of melatonin,as a neuro hormone capable of effectively alleviating social interaction deficits and regulating the development of dendritic spines,in Ctnnd2 deletion-induced nerve injury remains unclea r.In the present study,we discove red that the deletion of exon 2 of the Ctnnd2 gene was linked to social interaction deficits,spine loss,impaired inhibitory neurons,and suppressed phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt) signal pathway in the prefrontal cortex.Our findings demonstrated that the long-term oral administration of melatonin for 28 days effectively alleviated the aforementioned abnormalities in Ctnnd2 gene-knockout mice.Furthermore,the administration of melatonin in the prefro ntal cortex was found to improve synaptic function and activate the PI3K/Akt signal pathway in this region.The pharmacological blockade of the PI3K/Akt signal pathway with a PI3K/Akt inhibitor,wo rtmannin,and melatonin receptor antagonists,luzindole and 4-phenyl-2-propionamidotetralin,prevented the melatonin-induced enhancement of GABAergic synaptic function.These findings suggest that melatonin treatment can ameliorate GABAe rgic synaptic function by activating the PI3K/Akt signal pathway,which may contribute to the improvement of dendritic spine abnormalities in autism spectrum disorders.

Hepatoprotective effects of Xiaoyao San formula on hepatic steatosis and inflammation via regulating the sex hormones metabolism

BACKGROUND The modified Xiaoyao San(MXS)formula is an adjuvant drug recommended by the National Health Commission of China for the treatment of liver cancer,which has the effect of preventing postoperative recurrence and metastasis of hepatocellular carcinoma and prolonging patient survival.However,the molecular mechanisms underlying that remain unclear.AIM To investigate the role and mechanisms of MXS in ameliorating hepatic injury,steatosis and inflammation.METHODS A choline-deficient/high-fat diet-induced rat nonalcoholic steatohepatitis(NASH)model was used to examine the effects of MXS on lipid accumulation in primary hepatocytes.Liver tissues were collected for western blotting and immunohisto chemistry(IHC)assays.Lipid accumulation and hepatic fibrosis were detected using oil red staining and Sirius red staining.The serum samples were collected for biochemical assays and NMR-based metabonomics analysis.The inflammation/lipid metabolism-related signaling and regulators in liver tissues were also detected to reveal the molecular mechanisms of MXS against NASH.RESULTS MXS showed a significant decrease in lipid accumulation and inflammatory response in hepatocytes under metabolic stress.The western blotting and IHC results indicated that MXS activated AMPK pathway but inhibited the expression of key regulators related to lipid accumulation,inflammation and hepatic fibrosis in the pathogenesis of NASH.The metabonomics analysis systemically indicated that the arachidonic acid metabolism and steroid hormone synthesis are the two main target metabolic pathways for MXS to ameliorate liver inflammation and hepatic steatosis.Mechanistically,we found that MXS protected against NASH by attenuating the sex hormone-related metabolism,especially the metabolism of male hormones.CONCLUSION MXS ameliorates inflammation and hepatic steatosis of NASH by inhibiting the metabolism of male hormones.Targeting male hormone related metabolic pathways may be the potential therapeutic approach for NASH.

A DiGeorge Syndrome Case Report—Challenges of Diagnosis and Management

Background: DiGeorge syndrome (also known as velo-cardio-facial syndrome) is a rare multisystem genetic disorder occurring in approximately 1 in 4000 to 1 in 6000 live births [1]. Although advances in genetic screening have improved diagnosis in developed countries, the condition remains underdiagnosed in developing nations such as the Republic of Moldova, where access to genetic testing and family planning services is limited. Routine prenatal screening usually includes regular ultrasounds, monitoring of blood pressure, complete blood counts, coagulation studies, glucose, urine protein, and urine culture. Current ultrasound techniques have limitations in detecting this syndrome due to variability in interpretation, and genetic testing is often performed based on clinical discretion. The ultrasound could potentially point towards a genetic problem, as in DiGeorge, if multiple cardiac malformations are spotted in utero, but most cases such as this one are diagnosed after birth while being described as totally normal on prenatal ultrasound. Purpose: This study aims to highlight the diagnostic challenges and the need for comprehensive evaluation in identifying DiGeorge syndrome, emphasizing the importance of considering the syndrome as a whole rather than focusing on isolated organ system issues. Method: We present a case report of a 6-month-old girl who, after an uneventful pregnancy and normal prenatal ultrasound, presented with cardiac insufficiency. Following extensive investigations and multiple surgical interventions, DiGeorge syndrome was diagnosed at 9 months of age. Results: The patient’s diagnosis was delayed due to the lack of prenatal markers and the reliance on separate investigations of affected organ systems. Despite several interventions aimed at managing her symptoms, the final diagnosis was made after observing the association of multiple clinical features and conducting comprehensive genetic testing. Conclusions: This case underscores the importance of a holistic approach to diagnosis, which involves a thorough patient history, integration of diverse diagnostic tests, and recognition of the syndrome’s multi-system nature. It highlights the necessity for improved diagnostic protocols and increased awareness in regions with limited access to advanced genetic testing to prevent delays in identifying DiGeorge syndrome and to facilitate timely and appropriate management.

Effects of Homologous Genome 7 on Gas Exchange and Chlorophyll Fluorescence Parameters of Wheat

[Objective]To study the effects of homologous chromosomes 7 （7A,7B and 7D） on wheat photosynthesis and provide theoretical basis for breeding high photosynthetic efficiency wheat by genetic,physiological and biochemical means. [Method]The Triticum asetivum cultivar Chinese Spring wheat and nullisomic wheat （N7A,N7B and N7D） were planted in greenhouse. The photosynthetic indexes were determined at early filling stage. [Result]The photosynthetic rate （Pn）,stomatal conductance （Gs）,primary photochemical efficiency （Fv/Fm）,actual chemical efficiency of photosystem II （ФPS II） and apparent electron transfer rate of photosystem II （ETR） were significantly lower in the N7A and N7B than in the Chinese Spring （P0.05）. The photosynthetic rate and stomatal conductance was significantly lower in the N7D than in the Chinese Spring （P0.05）. The Fv/Fm,ФPS II,ETR of N7D were higher than that of the Chinese Spring without significant difference. [Conclusion]The homologous chromosomes 7A and 7B have positive effects on photosynthetic rate and have relationship with stomatal conductance and photoreaction （chlorophyll fluorescence parameters）. The homologous chromosome 7D has negative effects on photosynthetic rate,which is mainly related to stomatal conductance rather than photoreaction.

Cx43、PTEN在卵巢上皮性肿瘤组织中的表达及临床意义

卵巢癌是女性生殖系统肿瘤中死亡率最高的肿瘤,由于卵巢组织的解剖与内分泌的复杂性使卵巢癌的早期诊断极为困难因此,寻找卵巢恶性肿瘤早期诊断指标已经成为研究热点。间隙连接蛋白43（connexin 43,Cx43）为一种细胞间隙连接蛋白,多种肿瘤细胞中都有间隙连接蛋白43表达的下降与缺失[1]。同源磷酸酶-张力蛋白（phosphatase and tensin homolog deleted on chromosome ten,

Molecular Cloning and Construction of agp Gene Deletion-mutant in Cyanobacterium Synechocystis sp. PCC 6803

The agp gene encoding the ADP-glucose pyrophosphorylase involved in cyanobacterial glycogen synthesis was amplified by PCR. The resulting agp fragment was cloned in plasmid pUC118 to generate plasmid pUCA. Part of the fragment within the agp DNA was deleted and replaced by an erythromycin resistance cassette to generate plasmid pUCAE, which was used to transform the Synechocystis sp. PCC 6803 wild-type strain and a mutant with resistance to erythromycin was obtained. PCR analysis of the genomic DNA from the resulting mutant indicated that the appropriate deletion and insertion indeed had occurred. The cell growth and Chl a, glycogen content in the mutant showed difference from those in the wild-type strain. The obtained biomass as well as the Chl a content in the mutant strain was higher than that of the wild-type strain, which suggested that the photosynthesis efficiency in the agp(-) strain was higher than that in the wild-type strain. No glycogen was found in the mutant, providing evidence for the correction of the mutant in physiological level.

Detection of Serum Aberrant CDKN2/P16 DNA in Colorectal Cancer

Objective： To search for a biomarker for colorectal cancer. Methods： The MSP, SSCP and deletion tests with serum have been taken simultaneously in 100 cases of colorectal cancer and 2 groups of controls, as well as the specimens of 26 cancer tissues and 22 paracancerous tissues and 29 cases of benign disease tissues for a contrast. Results： The aberrant methylation rate of P16 in the serum was 69.00%, deletion rate 4.00% and suspicious point mutation rate 15.00% in colorectal cancer patients. The data of cancer tissues were the same as those of the serum, but in paracancerous tissue those were significantly lower. In 10 cases, sequencing analysis revealed that there were 3 cases of missense, one case of frameshift and one case of nonsense. Among them, four cases had P16 protein deletion. As a tumor marker, the sensitivity of combined use of three methods was 88.00%, specificity 96.87% and accuracy 90.15%. The combined use of MSP and SSCP could obtain the same results. Conclusion： The content of DNA in serum is minimal, but it reflects the tumor burden of patients. The 10^-3 fragments of DNA could be detected in the serum by MSP. It can be used in the clinical diagnosis or popular investigation, and long-term postoperative follow-up.

分布式数据库同步更新的实现方法

把分支机构的数据库作为出版者和分发者,把上级中心的数据库作为订阅者,首先在分发者数据库中对表建立快照代理、日志阅读代理、分发代理,和存储过程的复制事物,并在数据库中记录同步状态信息,然后分发者连接出版者,阅读出版物的事务日志,搜索出带有复制标志的INSERT、UPDATE、DELETE语句和其他更新事务提交给订阅者,当分支机构数据发生更新,日志读取代理即时将更新信息推到上级单位中心的数据库中,实现分发者和订阅者数据库数据两者数据的一致性,从而实现分布式数据库数据的同步更新。

C++动态内存分配研究

本文介绍了C++中内存的分配方式及动态内存分配中常见的内存错误及处理方法.

中国南方汉族人群KIR基因多态性的研究

目的探究中国南方汉族人群自然杀伤细胞免疫球蛋白样受体(KIR)基因的多态性及基因型和单体型特点,为KIR及KIR-HLA组合型多态性与疾病的关联研究提供基础数据资料。方法采用PCR-SSP方法对503名非亲缘关系的南方汉族个体进行KIR基因检测,依据Allele Frequency Net Database的国际标准对其进行基因型和单体型分析。根据第5外显子是否存在22bp碱基缺失,2DS4进一步分为2DS4-normal和2DS4-deleted等位基因。结果在检测的503份标本中,检出了目前已知的所有KIR框架基因,

Foxp3-6054基因多态性与变应性结膜炎的关系

目的探讨Foxp3-6054(deletion/ATT,rs5902434)基因与尘螨变应性结膜炎遗传易感性的相关性。方法应用PCR-SSP技术为63例无亲缘关系的尘螨变应性结膜炎患者和100例无血缘关系的健康汉族人行Foxp3-6054(deletion/ATT,rs5902434)基因分型。结果变应性结膜炎组的Foxp3-6054(deletion/ATT,rs5902434)基因型频率与对照组相比,差异无统计学意义(P>0.05)。结论 Foxp3-6054(deletion/ATT,rs5902434)位点基因多态性与变应性结膜炎不相关。

Studies on Crystalline Growth of MoFe Protein from a nifZ Deleted Strain of Azotobacter vinelandii

Under a given condition of crystallization, dark brown short rhombohedron crystals could be obtained from Δ nifZ MoFe protein purified from a nifZ deleted mutant strain of Azotobacter vinelandii Lipmann. Systematic studies on the effect of concentrations of PEG 8000,MgCl 2, NaCl,Tris and buffer pH on the crystallization and crystal growth of the protein showed that the protein could not be crystallized in lower concentrations of the chemicals and lower buffer pH. A large amount of smaller crystals of the protein appeared in a week with gradual increasing in the chemical concentrations and pH≥8.0. When the chemical concentrations were further increased, the time for crystallization was increased and a few high grade crystals of larger size were formed. If the concentrations of the chemicals were continuously increased, many crystals with smaller size, and, sometimes of poor quality appeared again and eventually ceased to produce any crystals. The optimal concentration for each of the above mentioned chemicals varies with other variable factors. Only one bigger crystal (both of the longest two sides: 0.16 mm) could be obtained in a hanging drop of protein sample when the concentrations of PEG 8000, MgCl 2, NaCl,Tris and protein were kept at 1.86%, 300 mmol/L, 400 mmol/L, 53 mmol/L and 4.64 g/L , respectively, with Tris buffer pH 8.2.

乳腺癌PTEN和S100A4表达的临床病理学意义

乳腺癌是女性最常见的恶性肿瘤之一,目前,乳腺癌的病因尚未完全清楚,其发生发展与多种癌基因和抑癌基因异常改变密切相关。S100A4和PTEN（phosphatase and tensinhomolog deleted on chromosome 10）与乳腺癌的关系成为近年研究热点。

MySQL触发器在电子商城系统中的应用

触发器是一种特殊的存储过程,主要通过事件触发从而执行。通过一个订单与库存管理的案例,详细阐述了insert、delete、update触发器的特性及在电子商城系统中的应用。

三种内存泄漏检测方法的比较

介绍了三种内存泄漏的检测方法:第一种是利用各种检测工具;第二种是通过重载new和delete操作符;第三种是奇异的循环模板模式对象计数法,它们都是程序在运行状态下的检测方法。最后文章比较了它们的优缺点。

控制C++的内存分配

内存分配运算符new/delete是为通用目的设计的,在特殊情况下,它不能满足我们的需要。本文给出了当需要动态分配大量的但很小的对象,或者程序必须在有限的资源情况下运行很长时间而不允许出现堆内存耗尽或出现很多碎片的解决方案。

SQL Server 2005触发器在数据库开发中的应用

在保持数据完整性和一致性方面,触发器起了很重要的作用。文章通过对单次操作多次触发代码的改进,解决了在实际操作过程中对大量记录进行更新和删除操作时,保留操作轨迹的表格不能完全记录这些操作的问题。

flush_action重选参数设置对数据业务用户感知的影响分析与设置优化

日常网络优化过程中往往只着重对语音无线参数进行调整优化,但随着数据业务突飞猛进的发展,GPRS承载网中同样存在设备参数的调整和优化问题。大家都知道在语音业务中用户从原来的小区进入到新的小区要进行切换,而数据业务中不存在切换流程,只有对应的小区重选,小区重选参数设置不合理会直接影响数据业务传输速率甚至会导致传输过程失败,从而对用户感知度产生较大影响,因此,合理优化小区重选参数就显得尤为重要,本文主要要对flush-ll-ack消息参数action设置不同对网络时延造成的不同影响进行论述分析,从而通过该参数调整优化来进一步提升数据业务网络质量,提升用户感知度。