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Endoscopic ultrasonography (EUS) and magnifying endoscopy (ME) reliably determine indications for endoscopic resection in patients with superficial esophageal squamous cell carcinoma (SESCC). ME is widely accepted for predicting the invasion depth of superficial esophageal cancer with satisfying accuracy. However, the addition of EUS is controversial.To evaluate the diagnostic efficiency of MEWe retrospectively analyzed patients with suspected SESCC who completed both ME and EUS and then underwent endoscopic or surgical resection at Sun Yat-Sen University Cancer Center between January 2018 and December 2021. We evaluated and compared the diagnostic efficiency of EUS and ME according to histological results, and investigated the influencing factors.We included 152 lesions from 144 patients in this study. The diagnostic accuracies of ME and EUS in differentiating invasion depth were not significantly different (73.0% and 66.4%,Preoperative diagnosis of SESCC should be conducted endoscopically using white light and magnification. In China, EUS can be added after obtaining patient consent. Use of a high-frequency miniature probe or miniature probe combined with conventional EUS is preferable.

Alzheimer's disease (AD) is a multifactorial disease, and it has become a serious health problem in the world. Senile plaques (SPs) and neurofibrillary tangles (NFTs) are two main pathological characters of AD. SP mainly consists of aggregated β-amyloid (Aβ), and NFT is formed by hyperphosphorylated tau protein. Sleep-wake disorders are prevalent in AD patients; however, the links and mechanisms of sleep-wake disorders on the AD pathogenesis remain to be investigated. Here, we referred to the sleep-wake disorders and reviewed some evidence to demonstrate the relationship between sleep-wake disorders and the pathogenesis of AD. On one hand, the sleep-wake disorders may lead to the increase of Aβ production and the decrease of Aβ clearance, the spreading of tau pathology, as well as oxidative stress and inflammation. On the other hand, the

Glutamic acid (Glu) is not only an important protein building block, but also a signaling molecule in plants. However, the Glu-boosted thermotolerance and its underlying mechanisms in plants still remain unclear. In this study, the maize seedlings were irrigated with Glu solution prior to exposure to heat stress (HS), the seedlings' thermotolerance as well as osmoregulation, glyoxalase, and non-glyoxalase systems were evaluated. The results manifested that the seedling survival and tissue vitality after HS were boosted by Glu, while membrane damage was reduced in comparison with the control seedlings without Glu treatment, indicating Glu boosted the thermotolerance of maize seedlings. Additionally, root-irrigation with Glu increased its endogenous level, reinforced osmoregulation system (i.e., an increase in the levels of proline, glycine betaine, trehalose, and total soluble sugar, as well as the activities of pyrroline-5-carboxylate synthase, betaine dehydrogenase, and trehalose-5-phosphate phosphatase) in maize seedlings under non-HS and HS conditions compared with the control. Also, Glu treatment heightened endogenous methylglyoxal level and the activities of glyoxalase system (glyoxalase I, glyoxalase II, and glyoxalase III) and non-glyoxalase system (methylglyoxal reductase, lactate dehydrogenase, aldo-ketoreductase, and alkenal/alkenone reductase) in maize seedlings under non-HS and HS conditions as compared to the control. These data hint that osmoregulation, glyoxalase, and non-glyoxalase systems are involved in signaling molecule Glu-boosted thermotolerance of maize seedlings.

Endoplasmic reticulum stress (ERS) and mitochondrial dysfunction have been suggested to relate with the pathology of Alzheimer's disease (AD). However, their cross-talk is needed to investigate further. Mitofusin-2 (Mfn2) is a member of mitochondria-associated membrane (MAM), which connects endoplasmic reticulum (ER) and mitochondria. This study investigated the protective effect of curcumin on thapsigargin (TG)-induced ERS and cell apoptosis and the role of Mfn2 on mitochondrial dysfunction. The cell viability of SH-SY5Y cells was decreased and cell damage and apoptosis were increased in a concentration-dependent manner when cells were treated with TG. TG upregulated the protein levels of GRP78, pSer981-PERK, and pSer51-eIF2α. Curcumin attenuated TG-induced damage on cell viability and apoptosis and downregulated the protein levels of GRP78, pSer981-PERK, and pSer51-eIF2α. TG caused the increases in intracellular reactive oxygen species (ROS) and in the protein levels of pSer40-Nrf2 and hemoglobin oxygenase 1 (HO-1). Curcumin decreased the TG-induced intracellular ROS but did not alter the protein levels of pSer40-Nrf2 and HO-1. TG resulted in the upregulation on Mfn2 expression and mitochondrial spare respiratory capacity but the downregulation on mitochondrial basal respiration and ATP production. Curcumin attenuated the TG-induced Mfn2 expression and mitochondrial stress. When Mfn2 was silenced by shRNA interference, curcumin failed to recovery the TG-damaged mitochondrial function. In general, the TG-induced ERS trigged mitochondrial dysfunction and cell apoptosis. Curcumin attenuates TG-induced ERS and the cell damage and apoptosis. Mfn2 is required for curcumin's protection against the TG-induced damage on mitochondrial functions.

Nitric oxide (NO) and hydrogen sulfide (H2S) are novel signaling molecules, which participate in plant growth, development, and response to stress. In this study root-irrigation with 0.15 mM sodium nitroprusside (SNP, NO donor) up-regulated gene expression of L-CYSTEINE DESULFHYDRASE1 (LCD1), activities of L-cysteine desulfhydrase (LCD) and D-cysteine desulfhydrase (DCD), as well as an endogenous H2S level, compared to control seedlings. The SNP-up-regulated effects were enhanced by 0.5 mM sodium hydrosulfide (NaHS, H2S donor), but weakened by NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) and H2S scavenger hypotaurine (HT) alone. NaHS had no significant effect on gene expression and activity of nitrate reductase (NR, a NO candidate producing enzyme). These data indicate that NO could trigger the LCD/H2S signaling pathway in maize seedlings. To further investigate the effect of NO and H2S crosstalk on thermotolerance in maize seedlings, thermotolerance parameters and reactive oxygen species (ROS)-scavenging system were estimated. The results show that SNP increased survival rate and tissue viability, decreased malondialdehyde (MDA) accumulation, and electrolyte leakage in maize seedlings under heat stress (HS), implying NO could improve thermotolerance in maize seedlings. The NO-improved thermotolerance was impaired by H2S inhibitor DL-propargylglycine (PAG) and scavenger HT alone. Similarly, SNP up-regulated the gene expression of DEHYDROASCORBATE REDUCTASE (DHAR) and GLUTATHIONE REDUCTASE1 (GR1); activities of ascorbate peroxidase, glutathione reductase, and catalase; as well as levels of ascorbic acid, glutathione, flavonoids, carotenoids, and total phenols. SNP also reduced hydrogen peroxide and superoxide radical accumulation in maize seedlings under HS compared to the control. The effects of SNP on ROS and their scavenger system were weakened by PAG and HT alone. These data hint that NO could evoke thermotolerance in maize seedlings by triggering the LCD/H2S signaling pathway, and the ROS-scavenging system played a key role in the NO and H2S crosstalk-evoked thermotolerance.

Hydrogen sulfide (H2S) and methylglyoxal (MG) were supposed to be novel signaling molecules in plants. However, whether interplay between H2S and MG can initiate thermotolerance in maize seedlings and in relation to metabolism of reactive oxygen species (ROS) and osmolytes is little known. In this study, watering with MG and NaHS (H2S donor) alone or in combination elevated survival and tissue vigor of maize seedlings under heat stress and coped with an increase in the biomembrane injury (as indicated in membrane lipid peroxidation and electrolyte leakage). The above-mentioned effects were separately weakened by MG scavengers (N-acetyl cysteine: NAC; aminoguanidine: AG) and H2S inhibitor (DL-propargylglycine, PAG) and scavenger (hypotaurine, HT). These suggested that the interplay between H2S and MG initiated the thermotolerance in maize seedlings. The further data indicated that, under non-heat stress and heat stress conditions, MG and NaHS alone or in combination modulated ROS metabolism by regulating the activities of antioxidant enzymes (catalase, ascorbate peroxidase, guaiacol peroxidase, glutathione reductase, monodehydroascorbate reductase, and dehydroascorbate reductase) and the contents of non-enzymatic antioxidants (ascorbic acid, glutathione, flavonoids, and carotenoids) in maize seedlings. In addition, MG and NaHS alone or in combination also separately modulated the metabolism of osmolytes (proline, trehalose, glycine betaine, and total soluble sugar), H2S (L-cysteine desulfhydrase and O-acetylserine (thione) lyase), and MG (glyoxalase I, glyoxalase II, and MG reductase). These physiological effects also were separately impaired by NAC, AG, PAG, and HT. The current data illustrated that the interplay between H2S and MG initiated the thermotolerance in maize seedlings by modulating ROS, osmolyte, H2S, and MG metabolism.

Glutamic acid (Glu) is not only a protein amino acid, but also a signaling molecule, which takes part in various physiological processes in plants. Our previous study found that root-irrigation with Glu could improve the heat tolerance of maize seedlings by plant Glu receptor-like channels-mediated calcium signaling (Protoplasma, 2019; 256:1165-1169), but its molecular mechanism remains unclear. In this study, based on the our previous work, the maize seedlings were treated with 1 mM Glu prior to be exposed to heat stress (HS), and then the expression of genes related to related to methylglyoxal (MG)-scavenging and osmoregulation systems was quantified. The results showed that Glu treatment up-regulated the gene expression of

Alzheimer’s disease (AD) is a neurodegenerative disorder in the central nervous system, and this disease is characterized by extracellular senile plaques and intracellular neurofibrillary tangles. Amyloid-β (Aβ) peptide is the main constituent of senile plaques, and this peptide is derived from the amyloid-β protein precursor (AβPP) through the successive cleaving by β-site AβPP-cleavage enzyme 1 (BACE1) and γ-secretase. AβPP undergoes the progress of post-translational modifications, such as phosphorylation and glycosylation, which might affect the trafficking and the cleavage of AβPP. In the recent years, about 10 phosphorylation sites of AβPP were identified, and they play complex roles in glycosylation modification and cleavage of AβPP. In this article, we introduced the transport and the cleavage pathways of AβPP, then summarized the phosphorylation and glycosylation sites of AβPP, and further discussed the links and relationship between phosphorylation and glycosylation on the pathways of AβPP trafficking and cleavage in order to provide theoretical basis for AD research.

Antigen-specific tolerizing DNA vaccines are one of the most promising strategies for rheumatoid arthritis (RA) treatment. They act by inducing potent immune tolerance instead of generalized immunosuppression. Recently, we developed a novel antigen-specific tolerizing DNA vaccine pcDNA-CCOL2A1 coding for chicken type II collagen (CCII) and confirmed its potent therapeutic efficacy in an established rat model of collagen-induced arthritis (CIA). Here we report the prophylactic vaccination efficacy of a single 300 μg/kg dose of pcDNA-CCOL2A1 against CIA incidence, severity, and onset. CCOL2A1 transcripts were detected in the blood of CIA rats 14-42 days after intramuscular injection by 300 μg/kg pcDNA-CCOL2A1. The expression of CCOL2A1 transcripts increased quickly on day 21, peaked at day 28, and then gradually decreased thereafter. Importantly, a single prophylactic vaccination of pcDNA-CCOL2A1 14 days before CIA establishment significantly reduced CIA incidence and severity, deferred its onset, and was as efficacious as the current gold standard drug, methotrexate. The marked effects on CIA incidence and severity closely corresponded to the expression of CCOL2A1. Furthermore, prophylactic vaccination with pcDNA-CCOL2A1 markedly decreased serum content of anti-type II collagen (CII) immunoglobulin G (IgG) antibodies, induced Th1-to-Th2 and Tc1-to-Tc2 shifts, and decreased the percentages of CD4

Three new cyclopentenoneacrylic acid derivatives, trichodermacid A (

Three new cyclopentenoneacrylic acid derivatives, trichodermacid A (1), trichodermester A (2), and trichodermester B (3), together with thirteen known compounds, were isolated from an ethyl acetate extract of Trichoderma atroviride H548, a fungus derived from mangrove sediment. The structures of the new compounds were elucidated by spectroscopic methods including HR ESI-MS, 1H NMR, 13C NMR, and 2D-NMR techniques. The antifungal activity of the isolated compounds was evaluated against tea pathogenic fungus Pestalotiopsis theae. Trichodermester A (2) showed potent anti P. theae activity with MIC of 125 μg/disc, while the other compounds were inactive.

Effective treatment of rheumatoid arthritis can be mediated by native chicken type II collagen (nCCII), recombinant peptide containing nCCII tolerogenic epitopes (CTEs), or a therapeutic DNA vaccine encoding the full-length CCOL2A1 cDNA. As recombinant CCII (rCCII) might avoid potential pathogenic virus contamination during nCCII preparation or chromosomal integration and oncogene activation associated with DNA vaccines, here we evaluated the importance of propeptide and telopeptide domains on rCCII triple helix molecular assembly. We constructed pC- and pN-procollagen (without N- or C-propeptides, respectively) as well as CTEs located in the triple helical domain lacking both propeptides and telopeptides, and expressed these in yeast Pichia pastoris host strain GS115 (his4, Mut+) simultaneously with recombinant chicken prolyl-4-hydroxylase α and β subunits. Both pC- and pN-procollagen monomers accumulated inside P. pastoris cells, whereas CTE was assembled into homotrimers with stable conformation and secreted into the supernatants, suggesting that the large molecular weight pC-or pN-procollagens were retained within the endoplasmic reticulum whereas the smaller CTEs proceeded through the secretory pathway. Furthermore, resulting recombinant chicken type II collagen pCα1(II) can induced collagen-induced arthritis (CIA) rat model, which seems to be as effective as the current standard nCCII. Notably, protease digestion assays showed that rCCII could assemble in the absence of C- and N-propeptides or telopeptides. These findings provide new insights into the minimal structural requirements for rCCII expression and folding.

A novel on-line comprehensive two-dimensional liquid chromatography (2D-LC) method by coupling mixed-mode gel liquid chromatography (MMG-LC) with reversed phase liquid chromatography (RPLC) was developed. A mixture of 17 reference compounds was used to study the separation mechanism. A crude water extract of Flos Carthami was applied to evaluate the performance of the novel 2D-LC system. In the first dimension, the extract was eluted with a gradient of water/methanol over a cross-linked dextran gel Sephadex LH-20 column. Meanwhile, the advantages of size exclusion, reversed phase partition and adsorption separation mechanism were exploited before further on-line reversed phase purification on the second dimension. This novel on-line mixed-mode Sephadex LH-20 × RPLC method provided higher peak resolution, sample processing ability (2.5 mg) and better orthogonality (72.9%) versus RPLC × RPLC and hydrophilic interaction liquid chromatography (HILIC) × RPLC. To the best of our knowledge, this is the first report of a mixed-mode Sephadex LH-20 × RPLC separation method with successful applications in on-line mode, which might be beneficial for harvesting targets from complicated medicinal plants.

This study investigated the role of anthrax toxin receptor 2 (ANTXR2) gene polymorphisms and capillary morphogenesis protein 2 (CMG2) expression in susceptibility and pathogenesis to ankylosing spondylitis (AS) in the Han Chinese in Beijing. A case-control study was performed using 602 AS patient samples meeting the revised New York criterion and 619 matched controls from Han Chinese individuals. Nineteen single-nucleotide polymorphisms (SNPs) of ANTXR2 genes were selected and genotyped using the Sequenom iPlex platform. Real-time polymerase chain reaction and flow cytometry were performed to investigate the impact of SNP polymorphisms on ANTXR2 transcription and CMG2 expression, respectively. The association of variants with AS was examined with UNPHASED 3.1.5. A novel association was observed between AS and three SNPs in the ANTXR2 gene (rs4690127, rs6823031, and rs4333130; P = 0.004, 0.011, and 0.013, respectively), confirming the association between rs433130 and AS in the Han Chinese. The strongest haplotype association was observed with rs4690127-rs6823031-rs4333130 (P = 2.5 × 10(-4)). rs6534639 and rs4333130 showed a cis-interaction (P = 0.027) in AS. ANTXR2 messenger RNA (mRNA) expression was significantly higher in the AS group than in the control group (P = 0.039). CMG2 expression in the lipopolysaccharide (LPS)-stimulated group was significantly lower than that in the control group (P = 0.018). This study reports a novel association between ANTXR2 and AS in the Han Chinese. ANTXR2 genetic polymorphisms affect ANTXR2 mRNA transcription and CMG2 expression. The opposing results observed for ANTXR2 transcription and CMG2 expression suggest a complex role of ANTXR2 polymorphisms in AS pathogenesis.

On the basis of understanding the we media and crisis public relations (PR) and its relations, taking the cases of female attacks in Beijing Heyi Hotel as an example, through the investigation of the events in the fermentation process and the diffusion process of Weibo, this article analyzes the information dissemination channels, the speed of dissemination and the scope of dissemination from the we media era. On this basis, combined with the current situation of Heyi Hotel crisis PR and compared the trend of the events afterwards, this article analyzes the new situation and new challenges faced by hotel crisis RR in the current situation, and through investigation and analysis, sums up the handling procedures of hotel crisis public relations from the we media era and provides constructive suggestions for the future crisis management in the hotel industry.

To examine the neuroprotection of baicalin, a flavonoid compound derived from the dried root of Scutellaria baicalensis Georgi, on neurons.A rat PC12 cell line was used to study the neuroprotection and possible mechanisms of baicalin on H₂O₂-induced neuron damage.Three anti- and one pro-apoptosis genes in PC12 cells were examined. Cell apoptosis was induced by H₂O₂ and apoptotic rate was obtained by flow cytometry. MTT for cell viability, immunofluorescence microscopy for promoter activity and western blot for gene expression were also employed.Data of MTT reduction assay and flow cytometry revealed that viability loss and apoptotic rate were reduced by pre-treatment of PC12 cells with baicalin for 24 hours. Baicalin was also found to increase SOD, GSH-Px activities and to decrease MDA level. Results from Western blot and immunofluorescence microscopy showed baicalin increased the expressions of survivin, Bcl-2 and p-STAT3 and decreased caspase-3 expression which were attenuated by AG-490.The results point to the possibility of the neuroprotective effects of baicalin on neuronal apoptosis induced by oxidative stress and indicate that activation of the JAK/STAT signalling pathway might involve the anti-apoptotic effect of baicalin.

The plastidic caseinolytic protease (Clp) of higher plants is an evolutionarily conserved protein degradation apparatus composed of a proteolytic core complex (the P and R rings) and a set of accessory proteins (ClpT, ClpC, and ClpS). The role and molecular composition of Clps in higher plants has just begun to be unraveled, mostly from studies with the model dicotyledonous plant Arabidopsis (Arabidopsis thaliana). In this work, we isolated a virescent yellow leaf (vyl) mutant in rice (Oryza sativa), which produces chlorotic leaves throughout the entire growth period. The young chlorotic leaves turn green in later developmental stages, accompanied by alterations in chlorophyll accumulation, chloroplast ultrastructure, and the expression of chloroplast development- and photosynthesis-related genes. Positional cloning revealed that the VYL gene encodes a protein homologous to the Arabidopsis ClpP6 subunit and that it is targeted to the chloroplast. VYL expression is constitutive in most tissues examined but most abundant in leaf sections containing chloroplasts in early stages of development. The mutation in vyl causes premature termination of the predicted gene product and loss of the conserved catalytic triad (serine-histidine-aspartate) and the polypeptide-binding site of VYL. Using a tandem affinity purification approach and mass spectrometry analysis, we identified OsClpP4 as a VYL-associated protein in vivo. In addition, yeast two-hybrid assays demonstrated that VYL directly interacts with OsClpP3 and OsClpP4. Furthermore, we found that OsClpP3 directly interacts with OsClpT, that OsClpP4 directly interacts with OsClpP5 and OsClpT, and that both OsClpP4 and OsClpT can homodimerize. Together, our data provide new insights into the function, assembly, and regulation of Clps in higher plants.

To study the utilization situation of harm reduction services among drug users and to analyze the reasons of the drug users' absence in the services as well as their evaluation of the services quality based on the gender perspective, and then to provide advice on the improvement of harm reduction services in the future.Cross-sectional surveys were carried out in four cities of Yunnan Province. The information was collected from the drug users in the drug rehabilitation centers and communities through outreach workers. The utilization of methadone maintenance treatment (MMT) and needle exchange service, the reason of the drug users' absence in the services and the evaluation towards the convenience, acceptability, as well as services quality were all surveyed.The valid samples were 579 with sex ratio 50.8% (male) and 49.2% (female). Most of the respondents (56.3%) had an educational level of junior high school and 44.0% were single. There were 61.8% (358/579) of respondents who used to utilize MMT service with female participation rate being 57.5%, which was lower than the male one (P0.05). And there were 58.2% (223/383) of the respondents who used to exchange needles from the outreach workers or needle exchange points with female participation rate being 52.9%, which was lower than the male one (P0.05). The leading cause of the drug users' absence in the services was their lack of understanding about the services. Among the people who used to utilize the services, the evaluations of women were better than those of men.Drug users have a low awareness of harm reduction services and female drug users are lack of the utilization towards services. More publicity and concern on harm reduction services in the future as well as exploration of the services which better fit the traits of women and privacy protection are suggested.

To study the allelic characteristics of "homozygote" resulted from low resolution genotyping of HLA-Cw locus and to provide more precise typing data for clinical transplantation.Forty three related allogeneic hematopoietic stem cell transplantation(allo-HSCT) donors and patients with HLA-Cw * 03, Cw * 07 homozygote, which were the most common gene groups in Chinese population, identified by low resolution genotyping level were retyped by high resolution PCR-SSP genotyping method, and three dimensional structure modelling was also made by using a solely developed HLA three-dimensional matching software (HLA strucMark version 1.0) to evaluate the effect of differences between two mismatched alleles and its relationship with GVHD occurrence.The typing results of high resolution level demonstrated that the confirmed allelic homozygotes for Cw * 03 and Cw * 07 were 14% and 43%, respectively, while the others were all heterozygotes. The ambiguous typing results could be confirmed by family data study or by high resolution typing methods when there was no family data available, Three-dimensional modeling results of the mismatched alleles undetected in low resolution typing level indicated that family data study or high resolution PCR-SSP genotyping were important in preventing graft-versus-host disease.When HLA-Cw homozygote results were found in low resolution genotyping method, the results should be reconfirmed by family data study or by high resolution typing methods to provide precise typing results for avoiding severe graft-versus-host disease.

Commerce is an important part of national economy and it plays an influential role to national economic growth; our country (China) also pays more and more attention to commercial development, for example the commercial affairs department decided to start establishing works of the commercial network plans of cities in 2008. This article wants to explain the importance and present situation of commerce through the research to the law of commercial development and economic growth. The empirical study of this article mainly centers on the law of commercial scale and economic growth. By selecting national data and 4 sample provinces and cities' local commercial data-commerce and Gross Domestic Product rate, total retail sales of consumer goods, per capita Gross Domestic Product and other relative data, using the method of correlation analysis to verify the law of commercial scale and economic growth, and comparing our country's industrial structures with those in other countries, it draws an conclusion as follows: Our country's commercial scale and area commercial scale have both already developed to a relatively stable level, but this level is low, under such circumstance, if we want to increase commercial economic benefit further, we must adjust commercial internal structures and expand market demand or make use of other methods to promote commercial development.

The development of the service industry is a significant symbol for measuring the level of economy and the modernization. Promoting the service industry's development, especially the modern one's, is a strategic method for enhancing citizens'quality on economy and running. Meanwhile, it is an inevitable choice to realize social coordination and the sustainable development. In view of this point, it is necessary for us to analyze and study the inner law of service industry and then guide its further development. Regarding service industry as the study object, this paper discusses deeply about the law and trend of its structure on long-term evolving, which is be considered as basis on adjusting and optimizing the structure of service industry in Zhenjiang, so as to induce it to develop healthily. Generally speaking, adopting the comparative and dynamic analysis and the empirical research method, this paper draws the conclusion, which has a vital practical significance on understanding development law of service industry, hierarchical of internal structure and the readjustment and optimization of the economic structure ,economic development, and build an overall well-to-do society.

The aim of this study was to investigate the parameters of gene frequencies, haplotype frequencies and linkage disequilibrium of HLA-A, -B, -Cw in HLA classical I loci for Chinese Han population. HLA-A, HLA-B and HLA-Cw loci were genotyped in 1014 unrelated China people using low resolution PCR-SSP typing method, and their genetic parameters were analyzed by statistic methods. The results indicated that among all the detected HLA-I genes, A*02 (0.33), A*11 (0.24), B*15 (0.14), B*13 (0.13), Cw*03 (0.25) and Cw*07 (0.18) were the popular gene groups distributing in Chinese Han population, and A*02-B*46 (0.071), A*11-B*15 (0.051), A*02-Cw*01 (0.084), A*11-Cw*03 (0.079), B*46-Cw*01 (0.095) and B*13-Cw*03 (0.071) were the predominant haplotypes in Han population. Additionally, A*02-B*46, A*30-B*13, A*30-Cw*06, A*02-Cw*01, B*46-Cw*01 and B*58-Cw*03 were statistically significant with strong linkage disequilibrium. While A*02-B*15, A*02-B*40, A*24-Cw*03, A*02-Cw*03 and A*31-Cw*03 were in low linkage disequilibrium, among them A*24-Cw*03 appeared frequently in HLA recombination events. In addition, A*02-B*46-Cw*01 (0.075), A*30-B*13-Cw*06 (0.046), A*11-B*13-Cw*03 (0.045), A*33-B*58-Cw*03 (0.044), A*11-B*15-Cw*08 (0.027), A*02-B*38-Cw*07 (0.023) and A*11-B*40-Cw*07 (0.022) were the main extended haplotypes in Han population. In conclusions, this study investigated systematically the genetic polymorphism features of Chinese Han population, which may provide useful genetic parameters for researches in colonial evolution, clinical transplantation and disease susceptibility.

In order to investigate the expression of heavy chain of HLA-B * 2705 in prokaryotic system and identify its activity, the extra-membrane gene fragment of HLA-B * 2705 was amplified from full-length HLA-B*2705 cDNA by PCR and cloned into pGEM-T vector. After identification by sequencing, the prokaryotic expressing vector pET32a (+)-B * 2705 was constructed. The antigenic activity of expressed protein was identified by Western blot and antibody blocking reaction. The results indicated that the fused HLA-B * 2705 protein expression with high efficiency was obtained. The expressed product was more than 50% of the total bacteria protein. The antigenic activity of expressed protein was confirmed by Western blot and antibody blocking reaction. It is concluded that HLA-B * 2705 fusion protein are obtained as basis for the further studies.

To investigate the therapeutic effect of gene vaccine encoding chicken collagen type II (CC II) on collagen-induced arthritis (CIA) comprehensively.Three groups (CIA) were given a single intravenous injection of plasmid pcDNA-CCOL2A1 (20 microg/kg, 200 microg/kg, 400 microg/kg) respectively and one group (CIA) was injected 200 microg/kg pcDNA3.1 as a control. The effect of gene vaccine (pcDNA-CCOL2A1) was evaluated according to the arthritis score, radiological and histological examinations.The severity of arthritis of CIA rats which were administered 200 microg/kg pcDNA-CCOL2A1 was significantly reduced from the fifth day. According to the radiological and histological examinations, the articular cartilage as well as subchondral bone trabeculae are similar to those of the normal groups, so the bone and articular cartilage structure were protected after treatment with 200 microg/kg pcDNA-CCOL2A1 with a little synovial hyperplasia. The therapeutic effect of 200 microg/kg pcDNA-CCOL2A1 group has significant difference in comparison with that of the pcDNA3.1 group (P0.05) and the arthritis scores are reduced to about 50% of those in control groups, but 20 microg/kg group and 400 microg/kg group has no therapeutic effect in our observation (P0.05).The new gene vaccine pcDNA-CCOL2A1 has significant therapeutic effect on CIA rats, and the treatment may therefore be an effective strategy for RA patient clinically.

To analyze the distribution of genes in HLA-Cw locus from Han population of China in a large scale, and to provide basic data for further study on the genetic characteristics of HLA-Cw locus of this population.Totally 1285 unrelated Chinese Han individuals were typed by PCR-SSP, and statistics was utilized to investigate the distribution rules of detected genes.Twenty-three HLA-Cw alleles were identified in Chinese Han population, out of them HLA-Cw*01, *03, *07 and *08 were the commonest genes, which accounted for frequencies of 0.1529, 0.2385, 0.1747 and 0.1004, respectively. Five genes which could not be identified by serological method were deaed: HLA-Cw*12, *14, *15, *16 and *17. Hardy-Weinberg test showed that the observed genetic polymorphism distribution values were correspondent with the expected (chi-square=73.74, df=98, P0.5).This study may serve a full-scale scientific genetic parameters of HLA-Cw genes for Chinese Han population studies.

To investigate the recombination event occurring between HLA-B and -Cw loci discovered in a family of Chinese Han nationality with an acute myeloid leukemia (AML) patient.Peripheral blood samples were collected from a Chinese man with M5 type AML, aged 39, and his healthy wife and daughter, all of Han nationality. HLA class I (-A, -B, and -Cw) and II (-DRB1 and -DQB1) alleles were typed by both low and high resolution PCR with sequence specific primers (PCR-SSP) and sequence-based typing (SBT). Then the recombination sites were analyzed by family study.The 2 haplotypes of the patient, his daughter, and his wife were A*2402101-Cw*030401/0402-B*1301-DRB1*0406-DQR1*030302/0303 and A*02011-Cw*150201/0202-B*4002-DRB1*1405-DQB1*05031, A*02011-Cw*150201/0202-B*1301-DRB1*0406-DQB1*030302/0303 and A*2406-Cw*0602-B*1302_DRB1*070101/0102-DQB1*0202, and A*330301/0302-Cw*030201/0202-B*58-1-DRB1*17-DQB1*0202 and A*2406-Cw*0602-B*1302-DRB1*070101/0102-B*5801-DRB1*0202 respectively. Family study demonstrated that A*02011-Cw*150201/0202 recombination and B*1301-DRB1*030302/0303 recombination carried by the daughter came from the 2 isolated chromosomes of her father, indicating that the recombination event occurred between HLA-B and -Cw loci during meiosis of the father and resulted in a new HLA haplotype that was inherited by the daughter.An unusual HLA-B/Cw recombination event occurring between HLA-B and -Cw loci has been found in a Han family, which helps further study the mechanisms of HLA recombination.

This study was aimed to establish downstream purification procedure by which the protein of interest can be purified to higher purity rapidly and efficiently. The different combinations of various purification strategies, methods and conditions were compared, including reversed phase chromatography, metal chelating chromatography, anion exchange chromatography, blue dye affinity chromatography, filtration chromatography and so on. The results showed that in reversed phase chromatography, isolated protein of interest was denatured and precipitated immediately after chromatography because methanol or acetonitrile were adopted as the organic phase. In blue dye affinity chromatography expecting to purify the protein of interest in one step, protein of interest was difficultly differentiated from mixed protein as much proteins bound to the chromatography media by non-specific affinity. While there is a translation-enhancing sequence T7-g10 in the PRSETA-B7-2-PE40KDEL expression vector, so it adds 6 histidines to the N terminus of the protein of interest, this allows to purify the protein of interest by metal chelating chromatography. Based on this characteristic, a three-step chromatography line including metal chelating chromatography, anion exchange chromatography and filtration chromatography was finally established after repeated experiments. By this way the purity of protein of interest reached 95% and the total recovery rate was 8%. The result of Western blot indicated that the expressed and purified recombinant B7-2-PE40KDEL could specifically bind with mAb against human B7-2 and multiple antibody against PEA. The cytotoxicity of the recombinant toxin tested by MTT method showed that the B7-2-PE40KDEL could selectively kill Jurkat cell line expressing CD28 receptor well and had no killing effect on the Raji cell line unexpressing CD28 receptor. It is concluded that a high efficient and speedy three-step purification procedure for the purifying recombinant protein B7-2-PE40KDEL was established, and this procedure possess selective killing activity on CD28 positive T lymphocytes.

Understanding with greater clearness the characteristics of recombination within the human leucocyte antigen(HLA) is of deep significance to gaining an insight into the evolutionary process of shaping HLA allelic diversity and ultimately the human resistance against diverse pathogens. Family studies and statistical analysis of recombination have provided estimations of recombination fractions across the major histocompatibility complex and have identified the potential recombination hotspots. Other characteristics such as haplotype specificity and sequence motifs have been intensively studied. The recombination fractions, hotspots and other characters are reviewed in this paper.

To study the expression of recombinant human SCF-TPO fusion protein and its biological function.Four primers were designed according to known sequences of TPO and SCF. The functional amino acid domains of TPO and SCF were amplified by RT-PCR from fetus hepatocytes, respectively. The expression plasmid pET32a/SCF-TPO was constructed by VOE gene fusion technique and expressed in E. coli BL21(DE3) plysS as inclusion body after isopropyl-beta-D-1-thiogalactopyranoside (IPTG) induction. The fusion protein was tested by SDS-PAGE and Western blot. The biological functions of SCF-TPO fusion protein in MO7e cells was investigated by MTT method after purification with metal chelating chromatography.The high expression SCF-TPO fusion protein was obtained, reaching up to 30% of the total cellular protein. Western blot verified the correct fusion expression and MTT results showed the growth promoting effect of the SCF-TPO fusion protein on MO7e cells, with a higher promoting activity at 100 ng/ml.Expressed SCF-TPO fusion protein after renaturation has biological activity in promoting the proliferation of MO7e cells.

The objective was to identify some biochemical and physical properties for fusion protein IL6D24-PE40KDEL. Edman degradation, SDS-PAGE, peptide mass fingerprinting, Western blot and MTT were used for identification of the protein. The results showed that the sequence of N-terminus is Met-Ile-Asp-Lys-Gln-Ile, Met was added because of prokaryotic expression system; Western blot revealed that the purified protein could react with IL6 and PEA antibody. The purified protein IL6D24-PE40KDEL could kill the multiple myeloma cell lines U266 expressing high affinity IL6R, but it could not kill the cell lines CEM which not expressed IL6R; The molecular weight was 58.7 kD measuring by SDS-PAGE; peptide mass fingerprinting (PMF) confirmed that the construction of IL6D24-PE40KDEL was correct. A novel protein by Peptident database in EXPASY web site was identified. In conclusion, IL6D24-PE40KDEL is a new targeting protein with bioactivity of specific killing effect.

A novel recombinant interleukin6-Pseudomonas exotoxin fusion protein, having targeted cytotoxicity for leukemic cells highly expressing IL6R has been molecularly designed and constructed in this study.Firstly, REDLK at C-terminus of Pseudomonas exotoxin PE40 was replaced with KDEL using point mutagenesis technology. Secondly, a cDNA encoding interleukin-6 devoid of N-terminal 24 amino acids [IL6D24] was fused to 5' terminus of PE40KDEL DNA by the method of gene splicing by overlap extension, which could generate recombinant IL6D24-PE40KDEL and IL6D24-Linker-PE40KDEL fusion genes. Thirdly, recombinant fusion genes IL6D24-PE40KDEL and IL6D24-Linker-PE40KDEL were ligated into the EcoR I and Sma I cloning sites in the pBV220 plasmids respectively and then transformed into E.coli HB101 cells. The expressed recombinant exotoxin fusion proteins were purified to electrophoresis purity by Mono Q column chromatography. Its selectively killing was tested by the MTS colorimetric method using both U937 and CEM cells lines.Recombinant exotoxin fusion proteins IL6D24-PE40KDEL was expressed as a form of inclusion bodies at higher level of 40% approximately of total proteins in bacterial cells. Western blot showed that the purified products could react specifically with IL6 monoclonal antibody and PEA antiserum, respectively. IL6D24-PE40KDEL was selectively cytotoxic to U937 cells expressing IL6R-positive with ID50 of 250 ng/ml, and but not CEM cells expressing IL6R-negative.Two novel recombinant interleukin6-Pseudomonas exotoxin fusion proteins IL6D24-PE40KDEL and IL6D24-Linker-PE40KDEL have been successfully constructed and they can selectively kill the leukemic cells expressing highly IL6R.

To establish and stabilize a new HLA typing strategy, reference strand mediated conformation analysis (RSCA), and to conform its advantages by using HLA-A as target gene.20 standard DNA samples from international RSCA cooperative team were used to establish and stabilize RSCA. 84 DNA samples of related hematopoietic stem cell transplant donor-recipients were extracted from peripheral blood cells. HLA-A loci were typed both by RSCA and PCR-SSP.RSCA results of 20 standard samples were identical to those of international RSCA cooperative team, and the accuracy and replication rate were both 100%. Among the 84 samples, 82/84 (97.6%) cases could be designated definitely, and 33/84 (39%) cases could be typed to allelic level. In addition, 2/84 (2.4%) cases could not be detected by RSCA for their poor PCR results. 20 samples were randomly selected to identify the replication rate of RSCA, and the results demonstrated that the replication rate was 100%. Among the PCR-SSP typing results, there were about 10% samples needed to be typed for 1 - 3 times to confirm their types.RSCA has some advantages compared with the PCR-SSP typing method, which are high resolution, high sensitivity, high accuracy, high replication, capability to find new alleles, high throughput, low cost and suitability for unrelated hematopoietic stem cell transplant donor-recipient HLA typing. But there are still some defects in this new strategy, which are time-consuming for only one sample a time and higher DNA quality requirement. In addition, RSCA database needs to be further improved.

After search at hematopoietic stem cell banks for transplant donors, there may be several donors matched with given standards. To determine the most appropriate donor for a specific patient, the potential donors were analyzed and compared by three methods. The first is cross-reactive group (CREG) antigens, which defined as the public antigens that shared specific serological reaction patterns. The second is residue match theory, which concerned the three residues oriented upward toward the T-cell receptor. The third is comparing HLA three-dimensional structure models. The results of the three methods were not completely accorded in our case. However, some less matched donors could be excluded from the candidates and the range of selection was further reduced. It is concluded that combined application of three methods would contribute in selecting donor for hematopoietic stem cell transplantation in clinics.

Background The earlier and unprotected sexual behaviour of young Chinese, and the consequences of these actions, have become a health concern, posing a challenge to traditional Chinese concepts. The aim of the present study was to investigate changes in sexual behaviour and associated risks among adolescents and young people in mainland China over the past three decades. Methods: A meta-analysis was undertaken to comprehensively review the sexual behaviour of Chinese young people (aged from 10 to 24 years) over the past 30 years. Relevant data published from 1979 to 2009 in the Chinese literature database were identified and retrieved. Analysis was performed based on set criteria. Results: Seventy-five studies were identified that were published after 1990. Overall, the estimated prevalence of sexual intercourse among young people was 12.6%, with an average age at sexual debut of 19.4 years. The rate of condom use at sexual debut was 37.2%; 53.6% of young people reported not using a condom during the most recent act of sexual intercourse. The rates of unintentional pregnancy and abortion among the sexually active were 15.1% and 10.8%, respectively. Reviewing the data according to decades revealed that the number of young people engaging in sexual intercourse decreased from 14.3% in 1990–1999 to 11.8% in 2000–2009. However, these rates are higher than the prevalence of sexual activity reported before 1990, which, according to sporadic studies, did not exceeded 1%. Conclusions: There have been increases in sexual activity and high-risk sexual behaviour in Chinese young people in the decades since the adoption of the open door policy in China, particularly since the mid-1990s. High-risk sexual behaviours have contributed to certain adverse consequences in adolescents and young people, such as unwanted pregnancy and sexually transmissible infection. Thus, efficacious intervention strategies need to be developed and implemented.