Genetic characteristics of thalassemia in newborns in Baisha Li Autonomous County, Hainan Province / 中华地方病学杂志

Objective:To understand the incidence and genetic characteristics of thalassemia in newborns in Baisha Li Autonomous County, Hainan Province, and to provide data support for government decision-making departments to formulate appropriate policies for prevention and control of thalassemia.Methods:With the help of Newborn Disease Screening Network of Hainan Province, samples of dry blood spots on the heels of newborns born in Baisha Li Autonomous County from January to June 2020 were collected based on the principle of informed consent. Fluorescent PCR melting curve method was used to detect the common types of thalassemia genes in Chinese population, and some samples were verified by the PCR + flow-through hybridization method. Samples of suspected new or rare mutations were sent to gene companies for sequencing analysis.Results:A total of 391 samples of neonatal dry blood spots were collected, and 252 samples with thalassemia genes were detected, the detection rate was 64.45% (252/391). Among them, 213 samples with α-thalassemia genes were detected, and the detection rate was 54.48% (213/391); 13 samples with β-thalassemia genes were detected, and the detection rate was 3.32% (13/391); 26 samples with α- and β-thalassemia genes were detected, and the detection rate was 6.65% (26/391). Among the above mentioned thalassemia genotypes, 1 case of rare type α-thalassemia -α 4.2/HKαα and 1 case of rare type β-thalassemia β CD39/β N were detected. According to ethnicity, 176 samples with thalassemia genes were detected in 238 Li samples, with a detection rate of 73.95% (176/238); 67 samples with thalassemia genes were detected in 137 Han samples, with a detection rate of 48.91% (67/137); 9 samples with thalassemia genes were detected in 16 other ethnic samples, with a detection rate of 56.25% (9/16). Conclusions:The detection rate of neonatal thalassemia genes is relatively high in Baisha Li Autonomous County, Hainan Province, and α-thalassemia is the most common. It is recommended that relevant government departments of Hainan Province should carry out genetic testing of neonatal thalassemia in Baisha Li Autonomous County as soon as possible to ensure the quality of life of the newborns.

Retrospective analysis of a ten-year screening project for G6PD deficiency in neonates in Hainan Province / 中华检验医学杂志

Objective@#To investigate the prevalence of glucose-6-posphate dehydrogenase (G6PD) deficiency and its gene mutations among neonates in Hainan Province.@*Methods@#The G6PD activity of dried blood spots of 914 520 neonates born from 2007 to 2016 was screened by fluorescence spot test in Hainan Province. The G6PD/6-glucose phosphate dehydrogenase (6GPD) ratio method was used to confirm the diagnosis of suspected specimens, and 3 012 of year 2016 dried blood spots of neonates with G6PD deficiency were genotyped using the multicolor probe-based fluorescence melting curve analysis.@*Results@#From 2007 to 2016, 36 314 positive cases were screened in 914 520 neonates. A total of 26 370 cases of G6PD deficiency were diagnosed with an incidence rate of 2.88%(26 370/914 520) in Hainan Province. The incidences of G6PD deficiency were 2.80%(21 688/774 555) in ethnic Han population, 3.45% (4 292/124 419) in ethnic Li population, 3.31%(212/6 401) in ethnic Miao population and 1.95%(178/9 145) in other ethnic groups. There were significant differences in the incidence of G6PD deficiency in ethnic Han population and ethnic Li population(χ2=161.261, P=0.000), ethnic Miao population(χ2=6.104, P=0.013) and other ethnic groups(χ2=24.283, P=0.000). A total of 13 mutation types were detected by gene detection in 3 012 confirmed cases of G6PD deficiency, of which c.1376 G>T, c.1388 G>A, c.95 A>G and c.1024 C>T mutations and related combinations accounted for approximately 91.74%. Two mutations outside 16 genotypes, c.86 C>T and c.1311 C>T, were found by gene sequencing.@*Conclusions@#The incidence of G6PD deficiency among newborns in Hainan Province is high, and there are ethnic and regional differences. The dominant genetic mutations in Hainan Province are c.1376 G>T, c.1388 G>A, c.95 A>G and c.1024 C>T.

Retrospective analysis of a ten-year screening project for G6PD deficiency in neonates in Hainan Province / 中华检验医学杂志

Objective:To investigate the prevalence of glucose-6-posphate dehydrogenase (G6PD) deficiency and its gene mutations among neonates in Hainan Province.Methods:The G6PD activity of dried blood spots of 914 520 neonates born from 2007 to 2016 was screened by fluorescence spot test in Hainan Province. The G6PD/6-glucose phosphate dehydrogenase (6GPD) ratio method was used to confirm the diagnosis of suspected specimens, and 3 012 of year 2016 dried blood spots of neonates with G6PD deficiency were genotyped using the multicolor probe-based fluorescence melting curve analysis.Results:From 2007 to 2016, 36 314 positive cases were screened in 914 520 neonates. A total of 26 370 cases of G6PD deficiency were diagnosed with an incidence rate of 2.88%(26 370/914 520) in Hainan Province. The incidences of G6PD deficiency were 2.80%(21 688/774 555) in ethnic Han population, 3.45% (4 292/124 419) in ethnic Li population, 3.31%(212/6 401) in ethnic Miao population and 1.95%(178/9 145) in other ethnic groups. There were significant differences in the incidence of G6PD deficiency in ethnic Han population and ethnic Li population(χ 2=161.261, P=0.000), ethnic Miao population(χ 2=6.104, P=0.013) and other ethnic groups(χ 2=24.283, P=0.000). A total of 13 mutation types were detected by gene detection in 3 012 confirmed cases of G6PD deficiency, of which c.1376 G>T, c.1388 G>A, c.95 A>G and c.1024 C>T mutations and related combinations accounted for approximately 91.74%. Two mutations outside 16 genotypes, c.86 C>T and c.1311 C>T, were found by gene sequencing. Conclusions:The incidence of G6PD deficiency among newborns in Hainan Province is high, and there are ethnic and regional differences. The dominant genetic mutations in Hainan Province are c.1376 G>T, c.1388 G>A, c.95 A>G and c.1024 C>T.

Analysis of inborn error metabolism in 277 children with autism spectrum disorders from Hainan / 中华医学遗传学杂志

OBJECTIVE@#To assess the value of dry blood spot tandem mass spectrometry for the diagnosis of autism spectrum disorder (ASD).@*METHODS@#Peripheral blood samples of 277 autistic children were collected. Their amino acid and carnitine profiles were detected by liquid chromatography tandem mass spectrometry. Urine samples of suspected patients were collected for verification by gas chromatography mass spectrometry. Blood samples were also taken for genetic testing.@*RESULTS@#Of the 277 children with ASD, 19 (6.9%) were suspected to be with inborn error of metabolism (IEM), which included 6 cases with amino acidemia, 9 with organic acidemia and 4 with fatty acidemia. Three cases of phenylketonuria, one case of homocysteinemia, one case of propionemia, one case of methylmalonic acidemia, one case of glutaric acidemia, one case of isovaleric acidemia, one case of argininemia, one case of citrullinemia I and four cases of primary carnitine deficiency were confirmed by genetic testing, which yielded an overall diagnostic rate of 5.1% (14/277).@*CONCLUSION@#Our result has provided further evidence for the co-occurrence of ASD and IEM. Tandem mass spectrometry has a great value for the diagnosis and treatment of ASD in childhood.

The screening and genetic analysis of primary carnitine deficiency in the minority municipalities in Hainan Province / 中华检验医学杂志

Objective To investigate the incidence of neonatal carnitine deficiency (PCD) in eight minority autonomous cities and counties in Hainan Province (Changjiang, Ledong, Dongfang, Baoting, Baisha, Qiongzhong, Wuzhishan and Lingshui). Methods A total of 18, 701 cases of newborn filter paper dried blood samples were collected from August 1, 2017 to July 31, 2018 in eight minority autonomous cities and counties in Hainan Province, including 10051 male infants and 8650 female infants. Tandem mass spectrometry and the non-derivatized multi-amino acid, carnitine and succinylacetone assay kits produced by PE were used to detect free carnitine (C0) and acylcarnitine (C2, C3, C16, C18, etc.).The carnitine spec-trum was reexamined with the recall of mothers and infants whose C0 was less than 10μmol/L. Blood sam-ples from those who were still low were sent to the Beijing McKinnon gene for genetic diagnosis, and the urine samples were sent to the Guangzhou Golden Field for urine gas chromatography. Results Among the 18701 newborns, 5 cases were diagnosed with PCD. The incidence of neonatal PCD was 2.67/10000 (5/18701). Two cases of Li mutation c.388GA, two cases of Han mutation P.R254X and one case of Miao mutation P. Y84N were confirmed by gene detection. The confirmed children were treated with L-carnitine to avoid metabolic disorders or myocardial and skeletal muscle damage. Conclusions The incidence of neonatal PCD is high in eight cities and counties in Hainan Province. The mutation sites of neonatal PCD are different in Li and Han. The tandem mass spectrometry screening of PCD can guarantee the quality of the birth population.

Analysis of inborn error metabolism in 277 children with autism spectrum disorders from Hainan / 中华医学遗传学杂志

Objective@#To assess the value of dry blood spot tandem mass spectrometry for the diagnosis of autism spectrum disorder (ASD).@*Methods@#Peripheral blood samples of 277 autistic children were collected. Their amino acid and carnitine profiles were detected by liquid chromatography tandem mass spectrometry. Urine samples of suspected patients were collected for verification by gas chromatography mass spectrometry. Blood samples were also taken for genetic testing.@*Results@#Of the 277 children with ASD, 19 (6.9%) were suspected to be with inborn error of metabolism (IEM), which included 6 cases with amino acidemia, 9 with organic acidemia and 4 with fatty acidemia. Three cases of phenylketonuria, one case of homocysteinemia, one case of propionemia, one case of methylmalonic acidemia, one case of glutaric acidemia, one case of isovaleric acidemia, one case of argininemia, one case of citrullinemia I and four cases of primary carnitine deficiency were confirmed by genetic testing, which yielded an overall diagnostic rate of 5.1% (14/277).@*Conclusion@#Our result has provided further evidence for the co-occurrence of ASD and IEM. Tandem mass spectrometry has a great value for the diagnosis and treatment of ASD in childhood.

Heat shock protein 27 enhances the inhibitory effect of influenza A virus NS1 on the expression of interferon-beta / 生物工程学报

Heat shock protein 27 (HSP27) is a member of the small heat shock proteins (sHSP) and has multiple functions, it also plays an important role in the life cycle of some viruses. To investigate the regulatory effect of HSP27 during influenza virus infection, we cloned and expressed human HSP27 in both prokaryotic and eukaryotic cells, and demonstrated that HSP27 interacted with influenza A virus NS1 protein both in vivo and in vitro. Luciferase assay showed that HSP27 inhibited the expression of interferon-beta (IFN-beta) in infected cells, and independent of its phosphorylation. Moreover, HSP27 enhanced the inhibitory effect of NS1 on the expression of IFN-beta. Further analysis indicated that HSP27 exerted the inhibitory effect probably through influencing MDA5 of the RIG-I like helicase (RLH) pathway. The results suggested that HSP27 play a role in the innate immunity of infected cells, contributed to our understanding of the regulatory effect of host factors during influenza virus infection.

Establishment of a mouse model of chronic pulmonary infection with Pseudomonas aeruginosa and analysis of its inflammatory response / 中华微生物学和免疫学杂志

Objective To research the chronic respiratory infectious diseases caused by Pseudomonas aeruginosa need to objectively the reflection of the real environment in the body, so established a mouse model of chronic pulmonary infection with Pseudomonas aeruginosa and analyzed it's inflammatory response. Methods Establishment of chronic bronchial infection mice model that were inoculated with Pseudomonos aeruginosa-laden agarose beads, and analyzed its inflammatory response by detected the cytokines and MMP-2, and a differential cell count and a total leukocytes count were performed as well. Results Pseudomonas aeruginosa had been detected after infection, and there were changes in pathological. Pulmonary inflammation appeared in 1 d and reached near baseline levels by 7 d after inoculation. It was verified that the peak of inflammatory reaction in Pseudomonas aeruginosa infection is in 2-3 d; the mice did have detectable levels of circulating matrix metalloproteinase-2 ( MMP-2 ) after infection with Pseudomonas aeruginosa. MMP-2 concentrations in the blood serum peaked at 3 d after inoculation. It is indicated that Pseudomonas aeruginosa can initiate a certain degree of pulmonary fibrosis on the basis of the pulmonary inflammation. Conclusion In this study, chronic bronchial infection animal model affected by Pseudomonas aeruginosa was established successfully. Base on this animal model, we can do the pathogenicity and drug resistance of Pseudomonas aeruginosa further study.

Biomechanical analysis of spinal implants / 中国组织工程研究

With development of modern medicine and tissue engineering,spinal implants have been commonly used in clinic for spinal diseases. According to post-implant functional features,the spinal implants are divided into static fixation system (pedicle screw,steel plate and intervertebral fusion) and dynamic fixation system (artificial intervertebral disc and artificial nucleus pulposus). In addition,biocompatibility and mechanics performances are involved in the system. Implant loosening and breakage are primary complications following surgery. Based on rich experience of treatment with various spinal implants and research advances of spinal biomechanics,implant selection is a complex problem and related to many factors. Thus,careful protocol before implantation,biomechanical factor evaluation and their relationships with patient occupation,sex,age and living habit are important.

Construction of eukaryotic expression plasmid carrying interleukin-18 receptor alpha,beta and establishment of cell line stably expressing interleukin-18 receptor beta / 中国组织工程研究

AIM:To construct human interleukin-18 receptor ?(IL-18R?) and interleukin-18 receptor ?(IL-18R?) eukaryotic expression vectors named pcDNA3.1-zeocin/IL-18R? and pcDNA3.0/IL-18R? respectively,and then stably transfect pcDNA3.0/IL-18R? into 293 cells in order to establish the cell line stably expressing IL-18R? and cell model which responds IL-18 signal transduction in vitro. METHODS:The experiment was performed at the laboratory of Department of Immunology,Peking University Health Science Center from June 2006 to May 2007. DNA encoding IL-18R? or IL-18R? was amplified and cloned into pcDNA3.1-zeocin or pcDNA3.0 vector,respectively. The recombinant of pcDNA3.0/IL-18R? was transfectd into 293 cells. After screening culture by G418,stable transfected cell line was established and the expression of IL-18R? was identified by Western-blot assay. The stable cell line with the combinant eukaryotic expression vector pcDNA3.1-zeocin /IL-18R? to establish the IL-18 signaling pathway,which was detected by NF-?B-dependent Luciferase. RESULTS:The eukaryotic expression vectors of IL-18R? and IL-18R? were constructed successfully. The gene of IL-18R? stably expressed in 293 cells and cell model of IL-18 signaling transduction in vitro could be established in these stable 293 cell lines. CONCLUSION:The establishment of NF-?B-activated cell model in vitro which responds to IL-18 can provide solid foundation for further experimental studies on IL-18 signaling transduction pathway.

Cloning, Expression, Purification of a Fusion Protein of SARS-CoV Nucleocapsid and Spike Protein Fragments and Its Immunological Characteristics / 中国生物化学与分子生物学报

The spike (S) and nucleocapsid (N) proteins, which are responsible for viral binding to cell surface receptors and the formation of ribonucleoprotein complexes during virion assembling, are major structure proteins of severe acute respiratory syndrome coronavirus (SARS-CoV). The expression of recombinant protein may give more accurate result for detecting SARS-CoV infection. A novel fusion protein,comprising of two fragments of N and S proteins from SARS-CoV, was prepared. Our computer-assisted analysis suggested that the immunodominant domains were located in the amino acid residues 1-227 of N protein and 450-650 of S protein, further the fusion of the two fragments did not change the immunochemical characteristics. The complementary DNA(cDNA) encoding N1-227 fused with S450-650 was obtained by sequence overlapping extension (SOE), and named NLS. It was cloned into pET-28a ( + ), an expression vector for His-tag fusion protein. This new constructed fusion protein was prokaryotic expressed in E. coli,and purified by metal chelate affinity chromatography with the purity over 95 %. The purified fusion protein was identified by anti-His monoclonal antibody and convalescence SARS patients serum. The NLS protein based ELISA showed that NLS maintained appropriate antigenicity and specificity to react with the sera of convalescent SARS patients. The functional NLS protein were successfully expressed and purified. And the fusion protein based ELISA can be used for detection of antibodies (Abs) against the S and N proteins of SARS-CoV. It may provided a novel diagnostic tool and have the potential application in developing of anti-SARS vaccine.

Studies on Extracting Process of Hujin Granules / 广州中医药大学学报

[Objective] To optimize the conditions of the extracting process of Hujin Granules. [Methods] The orthogonal design was applied. With the total emodin and the total anthraquinone (TA) content as the parameters for the alcohol-extraction, the concentration of alcohol, the volume of solvent and the extracting time were used for optimization of alcohol-extraction. With the total polysaccharide (TP) as the parameters for water-extraction, the soaking time, the volume of water and the extracting time were used for optimization of water-extraction. [Results] The optimum conditions of alcohol-extraction were: extracting with 70% alcohol 245 mL for 2 hours and extracting twice. The optimum conditions of water-extraction were: extracting with 80 mL water (not for soaking) for 1.5 hours, extracting 3 times. [Conclusion] The results indicate that the extracting process is rational and feasible, and can provide evidence for the extracting process of Hujin Granules.

Comparative Studies on the Anti- inflammatory and Analgesic Actions of Huang qi Guizhi Wuwu Decoction and Its Compositions / 中药新药与临床药理

Objective To investigate the anti- inflammatory and analgesic a ctions of Huangqi Guizhi Wuwu Decoction (HGWD) and its compositions and to study its rule of compatibility.Methods Analgesic action of HGWD and its compositio ns were observed by hot- plate method and acetic- acid- induced body twist me thod. Anti- inflammatory action of HGWD and its compositions were observed on t he models of xylene- induced auricular swelling in mice, celiac capillary perme ability in mice, cotton- induced granuloma in rats, albumem- induced arthritis and adjuvant arthritis in rats.Results HGWD and its drug pairs could inhibit the acute inflammation induced by albumen and xylene and rat adjuvant arthriti, decrease the celiac capillary permeability, inhibit the proliferation of granu loma, increase the pain threshold in mice and reduce the frequencies of body tw ist induced by acetic acid.Conclusion HGWD has significant anti- inflammatory and analgesic actions, so does single drug; Radix Astragali alone, but when Ra dix Astragali is used with the other drugs in HGWD, its effect can be enhanced.