Genotype-environment interaction on arterial stiffness: A pedigree-based study / 北京大学学报(医学版)

OBJECTIVE@#To utilized the baseline data of the Beijing Fangshan Family Cohort Study, and to estimate whether the association between a healthy lifestyle and arterial stiffness might be modified by genetic effects.@*METHODS@#Probands and their relatives from 9 rural areas in Fangshan district, Beijing were included in this study. We developed a healthy lifestyle score based on five lifestyle behaviors: smoking, alcohol consumption, body mass index (BMI), dietary pattern, and physical activity. The measurements of arterial stiffness were brachial-ankle pulse wave velocity (baPWV) and ankle-brachial index (ABI). A variance component model was used to determine the heritability of arterial stiffness. Genotype-environment interaction effects were performed by the maximum likelihood methods. Subsequently, 45 candidate single nucleotide polymorphisms (SNPs) located in the glycolipid metabolism pathway were selected, and generalized estimated equations were used to assess the gene-environment interaction effects between particular genetic loci and healthy lifestyles.@*RESULTS@#A total of 6 302 study subjects across 3 225 pedigrees were enrolled in this study, with a mean age of 56.9 years and 45.1% male. Heritability of baPWV and ABI was 0.360 (95%CI: 0.302-0.418) and 0.243 (95%CI: 0.175-0.311), respectively. Significant genotype-healthy diet interaction on baPWV and genotype-BMI interaction on ABI were observed. Following the findings of genotype-environment interaction analysis, we further identified two SNPs located in ADAMTS9-AS2 and CDH13 might modify the association between healthy dietary pattern and arterial stiffness, indicating that adherence to a healthy dietary pattern might attenuate the genetic risk on arterial stiffness. Three SNPs in CDKAL1, ATP8B2 and SLC30A8 were shown to interact with BMI, implying that maintaining BMI within a healthy range might decrease the genetic risk of arterial stiffness.@*CONCLUSION@#The current study discovered that genotype-healthy dietary pattern and genotype-BMI interactions might affect the risk of arterial stiffness. Furthermore, we identified five genetic loci that might modify the relationship between healthy dietary pattern and BMI with arterial stiffness. Our findings suggested that a healthy lifestyle may reduce the genetic risk of arterial stiffness. This study has laid the groundwork for future research exploring mechanisms of arterial stiffness.

The role of the high-level public health school in the development of the Center for Disease Control and Prevention / 中华流行病学杂志

The Ministry of Education and other four departments jointly issued the Notice on the Construction of high-level schools of public Health, proposing that "it will take ten years to build a number of high-level schools of public health, and form a high-quality education development system to adapt to the construction of modern public health system". At present, the construction of high-level public health schools in various universities in China is in full swing. The high-level School of Public Health and the CDC have played an important role in constructing the national public health system and the human health community. The high-level public health schools are of strategic significance and important value to the development of the CDC. The review presents reflections and insights on the role of high-level public health schools in the development of the CDC and the challenges they might face.

Progress in research of risk prediction of non-syndromic oral clefts using genetic information / 中华流行病学杂志

Non-syndromic oral cleft (NSOC), a common birth defect, remains to be a critical public health problem in China. In the context of adjustment of childbearing policy for two times in China and the increase of pregnancy at older childbearing age, NSOC risk prediction will provide evidence for high-risk population identification and prenatal counseling. Genome-wide association study and second generation sequencing have identified multiple loci associated with NSOC, facilitating the development of genetic risk prediction of NSOC. Despite the marked progress, risk prediction models of NSOC still faces multiple challenges. This paper summarizes the recent progress in research of NSOC risk prediction models based on the results of extensive literature retrieval to provide some insights for the model development regarding research design, variable selection, model-build strategy and evaluation methods.

Exploring the association between de novo mutations and non-syndromic cleft lip with or without palate based on whole exome sequencing of case-parent trios / 北京大学学报(医学版)

OBJECTIVE@#To explore the association between de novo mutations (DNM) and non-syndromic cleft lip with or without palate (NSCL/P) using case-parent trio design.@*METHODS@#Whole-exome sequencing was conducted for twenty-two NSCL/P trios and Genome Analysis ToolKit (GATK) was used to identify DNM by comparing the alleles of the cases and their parents. Information of predictable functions was annotated to the locus with SnpEff. Enrichment analysis for DNM was conducted to test the difference between the actual number and the expected number of DNM, and to explore whether there were genes with more DNM than expected. NSCL/P-related genes indicated by previous studies with solid evidence were selected by literature reviewing. Protein-protein interactions analysis was conducted among the genes with protein-altering DNM and NSCL/P-related genes. R package "denovolyzeR" was used for the enrichment analysis (Bonferroni correction: P=0.05/n, n is the number of genes in the whole genome range). Protein-protein interactions among genes with DNM and genes with solid evidence on the risk factors of NSCL/P were predicted depending on the information provided by STRING database.@*RESULTS@#A total of 339 908 SNPs were qualified for the subsequent analysis after quality control. The number of high confident DNM identified by GATK was 345. Among those DNM, forty-four DNM were missense mutations, one DNM was nonsense mutation, two DNM were splicing site mutations, twenty DNM were synonymous mutations and others were located in intron or intergenic regions. The results of enrichment analysis showed that the number of protein-altering DNM on the exome regions was larger than expected (P < 0.05), and five genes (KRTCAP2, HMCN2, ANKRD36C, ADGRL2 and DIPK2A) had more DNM than expected (P < 0.05/(2×19 618)). Protein-protein interaction analysis was conducted among forty-six genes with protein-altering DNM and thirteen genes associated with NSCL/P selected by literature reviewing. Six pairs of interactions occurred between the genes with DNM and known NSCL/P-related genes. The score measuring the confidence level of the predicted interaction between RGPD4 and SUMO1 was 0.868, which was higher than the scores for other pairs of genes.@*CONCLUSION@#Our study provided novel insights into the development of NSCL/P and demonstrated that functional analyses of genes carrying DNM were warranted to understand the genetic architecture of complex diseases.

Study on Thalassemia in Han Population in Sanya of Hainan Province / 中国实验血液学杂志

OBJECTIVE@#To study the distribution characteristics of thalassemia genotype in Han Population in Sanya of Hainan Province.@*METHODS@#Gap PCR and reverse dot hybridization were used to detect and analyze the thalassemia gene in 572 suspected thalassemia carriers of Han Population in Sanya.@*RESULTS@#Among the 572 Han Population in Sanya, 271 cases of thalassemia gene abnormality were detected, among which 161 cases were founded to be carriers of α-thalassemia gene. A total of 9 genotypes were detected, in the following order of the detection rate was －－SEA/αα，－α3.7/αα，－α4.2/αα，－－SEA/－α3.7，－－SEA/－α4.2，－α4.2/－α4.2，－α3.7/－α4.2，－α3.7/－α3.7，－－SEA/－－SEA. Among them, the deletion type (－－SEA/αα) in southeast Asia was the most common, accounting for 66 cases. 99 cases of β-thalassemia were detected, there were 7 genotypes, all of which were heterozygous. The order of the detection rate was CD41-42/βN, IVS-II-654/βN, CD17/βN, CD71-72/βN, -28/βN, －29/βN, CD27-28/βN. Among them, CD41-42/βN was the most common, accounting for 51 cases. In addition, 11 cases of combined α and β thalassemia were detected. Five kinds of genotypes were checked out, the order of detection rate was －α3.7/αα composite CD41-42/βN, －－SEA/αα composite IVS-II-654/βN, －α4.2/－α4.2 composite CD41-42/βN, －α4.2/αα composite －29/βN , －－SEA/ －α4.2 composite CD41-42/βN.@*CONCLUSION@#Han Population in Sanya of Hainan Province is a high-risk population of thalassemia, the genotype characteristics are different from other areas with high incidence of thalassemia in China. The main type of α-thalassemia is the deficiency mutation of southeast Asia, while CD41-42 heterozygous mutation is the main type of β-thalassemia.

Effect assessment of caries management by risk assessment in children in the Minhang district of Shanghai / 口腔疾病防治

Objective@#To assess the effect of caries management based on risk assessment in children and to provide the basis for the government to develop strategies to prevent and treat oral diseases.@*Methods@# From March 2018 to March 2019, 512 3-year-old children and 502 6-year-old children were selected by cluster sampling from kindergartens and primary schools, respectively, in the Minhang district of Shanghai, and oral examinations and questionnaires were carried out to assess baseline status. Then, the children were divided into 3 groups, including low-, middle- and high-risk groups, according to caries risk assessment. In each caries risk group, the children were randomly divided into an experimental group and a control group. The experimental groups were managed by risk assessment, and the control groups were provided basic oral public health services by the Shanghai government. The effect of caries prevention was evaluated 12 months later.@*Results@#Among the children with a high risk of caries, the incidence of caries was 51.22% in the 3-year-old control group, 34.17% in the experimental group, 51.27% in the 6-year-old control group and 33.15% in the experimental group, with statistical significance (P < 0.05). Among the children with a middle risk of caries, the incidence of caries was 38.71% in the 3-year-old control group and 7.32% in the experimental group, with statistical significance (P < 0.05). However, there was no significant difference in caries incidence between the control group and the experimental group for the children with low risk in the 3-year-old or 6-year-old groups and middle risk caries risk in the 6-year-old group. Both 3-year-old and 6-year-old children in the experimental group had a lower risk of caries than those in the observation group in 2019.@*Conclusion@#The caries prevention effect is remarkable in high-risk children, and caries management by risk assessment is probably recommended for children with high caries risk.

Study on variation trend of repetitive nerve stimulation waveform in amyotrophic lateral sclerosis / 中华医学杂志(英文版)

BACKGROUND@#Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease involving both upper and lower motor neurons with no effective cure. Electrophysiological studies have found decremental responses during low-frequency repetitive nerve stimulation (RNS) except for diffused neurogenic activities. However, the difference between ALS and generalized myasthenia gravis (GMG) in terms of waveform features is unclear. In the current study, we explored the variation trend of the amplitudes curve between ALS and GMG with low-frequency, positive RNS, and the possible mechanism is discussed preliminarily.@*METHODS@#A total of 85 ALS patients and 41 GMG patients were recruited. All patients were from Peking Union Medical College Hospital (PUMCH) between July 1, 2012 and February 28, 2015. RNS study included ulnar nerve, accessory nerve and facial nerve at 3 Hz and 5 Hz stimulation. The percentage reduction in the amplitude of the fourth or fifth wave from the first wave was calculated and compared with the normal values of our hospital. A 15% decrease in amplitude is defined as a decrease in amplitude.@*RESULTS@#The decremental response at low-frequency RNS showed the abnormal rate of RNS decline was 54.1% (46/85) in the ALS group, and the results of different nerves were 54.1% (46/85) of the accessory nerve, 8.2% (7/85) of the ulnar nerve and 0% (0/85) of the facial nerve stimulation, respectively. In the GMG group, the abnormal rate of RNS decline was 100% (41/41) at low-frequency RNS of accessory nerves. However, there was a significant difference between the 2 groups in the amplitude after the sixth wave.@*CONCLUSIONS@#Both groups of patients are able to show a decreasing amplitude of low-frequency stimulation RNS, but the recovery trend after the sixth wave has significant variation. It implies the different pathogenesis of NMJ dysfunction of these 2 diseases.

Quantitative determination of biogenic amine from Biomphalaria glabrata nervous system by UPLC MS/MS / 中国血吸虫病防治杂志

Objective To establish a method for the quantitative determination of serotonin and dopamine in the nervous sys-tem of Biomphalaria glabrata by using ultra high performance liquid chromatography-tandem quadrupole mass spectrometry (UPLC MS/MS). Methods The B.glabrata nervous system was broken in the pure methanol solution after obtaining it by dis-secting with microscope.Then,the supernatant containing the target substance after twice high speed centrifugation was got.The extraction was separated on an ACQUITY UPLC BEH Amide column with Waters TQ-XS series mass spectrometry detector, with ESI source and positive electrospray ionization mode when the machine testing. Results The detection limit of serotonin was 0.03 ng/ml and the limit of quantification was 0.1 ng/ml.The detection limit of dopamine was 0.05 ng/ml and the limit of quantification was 0.15 ng/ml.The recoveries of serotonin ranged from 90.68% to 94.72% over the range of 1 to 40 ng/ml.The re-coveries of dopamine ranged from 91.68% to 96.12% over the range of 1.0 ng/ml to 40 ng/ml. Conclusion The established UPLC MS/MS method is simple,stable and reproducible.It can be used for the quantitative analysis of serotonin and dopamine in the nervous system of B.glabrata snails.

Single Center Analysis of Pathogenic Characteristics of Lung Candida Infection and Other Pathogenic Bacteria Infection in the Patients with Hemotological Maliglancies in the Tropical Region of Hainan / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the infection characteristics of sputum and venous blood pathogen in the patients with hematological malignancies and respiratory symptom in the Hematology Department in tropical region and to investigate its drug-resistance so as to provide etiological evidence for clinical treatment.</p><p><b>METHODS</b>The pathogens and their drug-resistance of 2466 samples in the patients with hematological malignancies and respiratory symptom in the Department were analyzed retrospectively from January 2013 to November 2017. The samples were collected from sputum and venous blood.</p><p><b>RESULTS</b>The sputum sample culture in patients with hematologic diseases showed that 224 strains were isolated, out of them 98 strains (43.75%) were fungi mainly candida albicans (41 strains); and then 88 Gram-negative strains (39.28%), among them the main pathogenic bacteria were Escherichia coli(22 strains) and klebsiella Klebsiella pneumoniae(12 strains); and then 38 Gram-positive strains (16.96%), among them the main pathogeni-bacteria were Enteroccocus (14 strains) and Gram-positive bacilli (14 strains). The blood samples culture of patients with hematologic diseases showed that 61 strains were isolated, out of them the isolated rate of Gram-positive bactetia was higherst, which accounted for 55.74%(34/61), mainly including staphylococcus lominis (19 strains); and the isolated rate of Gram-negative bacteria was 44.26% (27/61), among them main pathogenic bacteria was Klebsiella pneumoniae (12 strains). The resistance test of pathogenic bacteria to drugs showed that the resistant rate of Gram-negative bacteria to tigecycline, imipenem and atl-962 duenner was lowest, while the Gram-positive pathogenic bacteria such as Enterococcus, Gram-positive bacilli and Staphylococcus aureus were sensitive to vancomycin, tigecycline and linezolid was high.</p><p><b>CONCLUSION</b>the patients in hematology department are infected easily in the hospital in tropical region. The main pathogens are fungal strains in the respiratory tract of patients with hematological malignancy according to the sputum culture results. The clinician in tropical regions should choose suitable antibiotics for anti-infective therapy, which is different from the situation in North China or other northern areas.</p>

Genotype Analysis of Patients with Thalassemia in Sanya Area of Hainan Province in China / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the frequency and spectrum of thalassemia gene mutations of the population in Sanya area of Hainan province in China.</p><p><b>METHODS</b>The type and frequency of gene mutation in 1060 patients with suspected thalassemia were analyzed by Gap-PCR and reverse dot blot (RDB).</p><p><b>RESULTS</b>The detection on mutation of thalassemia gene were found in 539 suspected thalassemia patients, the total detected rate was 50.85% (539/1060), out of them 330 (31.13%) were diagnosed with α-thalassemia, 162 (15.28%) with β-thalassemia, and 47 (4.43%) as carriers of both α and β-thalassemia. In α-thalassemia patients, genotype were as follows in proper order--SEA/αα (9.25%)、-α /αα (5.94%)，HbH (5.56%)，-α /αα (5.00%)，-α /-α (2.36%)，-α /-α (1.70%), and -α/-α(1.32%). In β-thalassemia patients, there were 9 gene mutations: CD41-42 (9.8%), CD17 (1.32%), 654 (1.23%), CD71-72 (1.23%), IVS-II-654 (1.04%), -28 (0.37%), CD43 (0.19%), -29 (0.18%) and βE (0.09%). In the α and β composite thalassemia there were 12 genotypes. The -α/αα was the most common genotype co-existed with β-thalassemia (1.70%), followed by the -α /αα genotype (0.94%).</p><p><b>CONCLUSION</b>The data of this study provide the frequency and the spectrum of thalassemia gene mutations in the sanya area of Hainan province, which can contribute to set up the strategies for the prevention and control of thalassemia in this area.</p>

Inhibition of the recombinant human endostatin adenavirus on experimental choroidal neovascularization in rats / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the inhibition of the recombinant human endostatin adenavirus (Ad-Es) on the experimental choroidal neovascularization(CNV) models by intravitreous injection.METHODS: Experimental CNV models were induced by semiconductor laser in 30 male Brown Norway(BN) rats and randomly divided into 3 groups with 10 rats in each group.At 21d after photocoagulation, the single administration group were given intravitreous injection with Ad-Es 0.01mL;the repeated administration group were given intravitreous injection with Ad-Es 0.01mL and a repeated injection 7d later;the saline control group were given intravitreous injection with saline 0.01mL.At 7d after final administration, the leakage of fundus fluorescein angiography (FFA) was observed.Various CNV areas were measured by using laser confocal microscopy of choroidal flatmount method.Pathology and ultrastructure were observed with light microscopy, the expressions of CD105 were measured by immunohistochemistry.RESULTS: The leakage of CNV of the administration group abviously decreased as compared with those in the saline group, the leakage of repeated administration group decreased compared with that of single administration group (P<0.05).Laser confocal microscope quantitative CNV analysis showed that CNV area of the administration group was significantly smaller than that of control group, the area of repeated administration group was smaller than that of single administration group (P<0.05).Under the light microscope, the vascular endothelial cell number and quantity of the administration group were significantly lower than that of the control group, the cell number of repeated administration group was lower than that of single administration group.CD105 expression of the administration group was significantly weaker than that in the saline group;the expression of repeated administration group was weaker than that of single administration group.CONCLUSION: Ad-Es can effectively inhibit semiconductor laser induced CNV in BN rats, and the inhibition effect of repeated administration group is better than that of single administration group.It may be a useful new method in the treatment of CNV.

Protection of Transcutaneous Acupoint Electrical Stimulation for Brain Injury Undergoing Intervention: a Clinical Observation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of transcutaneous acupoint electrical stimulation (TAES) combined dexmedetomidine on hemodynamic of intracranial aneurysmal subarachnoid hemorrhage patients undergoing intervention, and their protection for brain Injury.</p><p><b>METHODS</b>Totally 108 intracranial aneurysmal subarachnoid hemorrhage patients undergoing intervention were randomly assigned to the electroacupuncture (EA) group and the control group according to random digit table, 54 in each group. All patients were anesthetized with dexmedetomidine. Patients in the EA group were needled at bilateral Neiguan (PC6), Lieque (LU7), and Yunmen (LU2). Parameter setting was as follows: The dilatational wave at 1. 5 Hz, strength 2 - 4 mA, 30 min. Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), and heart rate (HR) were compared between the two groups immediately after entry into the room (T0), after administration (T1), intubating (T2), resuscitation (T3), extubation (T4), and leaving the operating room (T5). Levels of S100β protein (S100β) and neuron specific enolase (NSE) were compared between the two groups at T0, immediately after surgery (T6), 6 h after operation (T7), 12 h after operation (T8), and 24 h after operation (T9).</p><p><b>RESULTS</b>Compared with the same group at T0, SBP, DBP, MAP, and HR were significantly reduced in the two groups at T1-T5(P <0. 05), serum levels of S100β and NSE in the two groups were significantly increased at T6-T9 (P<0. 05). Compared with the control group at T1 - T5, SBP, DBP, MAP, and HR decreased in the EA group (P <0. 05). Compared with the control group at T6-T9, serum levels of S100β and NSE decreased in the EA group (P <0. 05).</p><p><b>CONCLUSION</b>TAES combined dexmedetomidine could effectively maintain stable hemodynamics of intracranial aneurysmal subarachnoid hemorrhage patients undergoing intervention, and regulate their serum levels of S100β and NSE.</p>

Chemical constituents in ethyl acetate fraction of Artemisia selengensis / 中草药

Objective: To study the chemical constituents in the ethyl acetate extract from Artemisia selengensis. Methods: The chemical constitunents were isolated by various column chromatographic methods. The structures of the compounds were elucidated on the basis of physiochemical properties and spectral analyses. Results: Eleven compounds were identified as trans-resveratrol (1), trans-cinnamic acid (2), caffeic acid (3), chlorogenic acid (4), gallic acid (5), luteolin (6), isorhamnetin (7), 7-methoxy coumarin (8), quercetin (9), acteoside (10), and 7-methoxy-4'-hydroxyisoflavone (11). Conclusion: Compounds 1-11 are isolated from A. selengensis for the first time.

Purification and characterization of a ribonuclease from the wild edible mushroom Armillaria luteo-virens.

A 15 kDa ribonuclease (RNase) was purified from dried fruiting bodies of the wild edible mushroom Armillaria luteo-virens. The simple 4-step purification protocol involved ion-exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion-exchange chromatography on SP-Sepharose and a final gel filtration by FPLC on Superdex-75. The RNase was unadsorbed on Affi-gel blue gel, but adsorbed on DEAE-cellulose and SP-Sepharose. The N-terminal amino acid sequence of purified RNase was AGVQYKLTILLV, which showed low sequence homology to those of previously reported RNases. The optimal pH and temperature of the enzyme were very close to 4.0 and 70°C, respectively. The enzyme showed considerably high ribonucleolytic activity and broad specificity towards polyhomoribonucleotides, with a specificity of poly(U)>poly(C)>poly (G)>poly(A). The ribonucleolytic activities towards poly(U), poly(C), poly(G) and poly(A) were 279.5, 184.1, 69.9 and 52.3 U/mg, respectively.

Exposure assessment on dietary lead and cadmium of residents in three areas of Zhejiang province from 2009 to 2010 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To assess the safety of dietary lead and cadmium intake in 3 areas of Zhejiang province.</p><p><b>METHODS</b>Using the total dietary study method, the study was conducted in 3 regions which represented coastal, city and rural areas in Zhejiang province from 2009 to 2010. The dietary survey was conducted on the residents (512 subjects) and the categories and volume of food consumption were obtained. The analytical food samples were obtained by food consumption survey, food aggregation, food sampling and preparation. The food samples were detected by inductively coupled plasma mass spectrometry (ICP-MS). The safety of dietary lead and cadmium intake was evaluated.</p><p><b>RESULTS</b>The median dietary lead intake (P₅₀) in Zhejiang province was 37.8 µg/d. The 97.5% dietary lead intake (P₉₇.₅) was 72.3 µg/d. The P₅₀ dietary lead intakes of different age and gender groups ranged from 23.2 to 44.2 µg/d. The P₉₇.₅ dietary lead intakes of different age and gender groups ranged from 34.2 to 88.1 µg/d. The P₅₀ dietary cadmium intake in Zhejiang province was 9.6 µg/d. The P₉₇.₅ dietary cadmium intake was 15.7 µg/d. The P₅₀ dietary cadmium intakes of different age and gender groups ranged from 6.4 to 11.4 µg/d, accounting 15.6% - 42.6% of PTMI (provisional tolerable monthly intake, 25 µg/kg). The P₉₇.₅ dietary cadmium intakes of different age and gender groups ranged from 10.5 to 21.4 µg/d, accounting 27.5% - 77.6% of PTMI. Vegetable (11.3 µg), cereal (11.0 µg) and meat (9.8 µg) were the first three food sources which accounted for 84.9% of dietary lead (P₅₀: 37.8 µg/d). Cereal (3.6 µg), vegetable (2.1 µg) and legume (0.9 µg) were the first three food sources which accounted for 68.8% of dietary cadmium (P₅₀: 9.6 µg/d).</p><p><b>CONCLUSION</b>Dietary lead and cadmium intakes of most residents in 3 areas of Zhejiang province as well as the average level are safe.</p>

Relationship between proteoglycans and proliferation of human salivary adencid cystic carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of down-regulated proteoglycans on the proliferation of human salivary adenoid cystic carcinoma (SACC).</p><p><b>METHODS</b>The short hairpin RNA (shRNA) plasmid silencing human xylosyltransferase-I (XT-I) gene was constructed and named shRNA-WJ3. Adenoid cystic carcinoma cells with high metastatic tendency (ACC-M) were transfected by shRNA-WJ3. The plasmid shRNA-HK not targeting any human gene was transfected into ACC-M cells used as negative control. After 48 h of transfection, the positive cells were screened by G418 to isolate the stable transfected cells. Real-time PCR and Western blotting were used to test the gene silence, and the proteoglycans contents of the cells were detected. The stable cell line silenced XT-I was named ACC-M-WJ3. MTT assay was performed to detect the cell proliferation. The cell cycle was analyzed by flow cytometry.</p><p><b>RESULTS</b>ShRNA-WJ3 showed powerful RNA interference and gene silence of XT-I. The inhibition rate was 83.70% of mRNA expression and 79.60% of protein expression respectively. The content of proteoglycans in ACC-M-WJ3 was down-regulated by 49.71%-54.59%. The results of MTT assay showed that the cell growth was inhibited significantly. S phrase decreased and G₁-G₀ phrase increased in group ACC-M-WJ3 compared with that of group ACC-M-HK (P < 0.05).</p><p><b>CONCLUSIONS</b>The down-regulated proteoglycans could inhibit the proliferation of human ACC-M cells.</p>

Immune effects of three different programs for revaccination among adults of non- and hypo-responders to hepatitis B vaccine / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the immune effects of three different programs for revaccination among adults of non- and hypo-responders to recombinant Hepatitis B vaccine.</p><p><b>METHODS</b>Those who were once immunized with recombinant Hepatitis B vaccine more than one standard schedule (0, 1, and 6 months) in two years and negative for Hepatitis B markers were randomly given three-different projects for revaccination. 34 adults of A group were given GM-CSF 300 microg by subcutaneous injection for the first day, then 10 microg each time by intramuscular route for routine immune method. 33 adults of B group were given Hepatitis B vaccine 20 microg each time. 33 adults of C group were given Hepatitis B vaccine 10 microg each time. The blood samples were collected before the first injection and in 1, 2 and 8 months following the first injection to test Anti-HBs.</p><p><b>RESULTS</b>At T1, the anti-HBs positive conversion rate of group A, B and C was 26.47%, 48.48% and 18.18% respectively (chi-2 = 7.20, P = 0.027). At T8, the anti-HBs positive conversion rate of group A (64.71%) and group B (75.76%) were higher than group C (39.39%), and there was significant difference (chi-2 = 9.07, P = 0.011). At T1, the anti-HBs level of group B (417.00 +/- 69.36) was higher than that of group A (203.74 +/- 79.56). At T2, the anti-HBs level of group B (458.17 +/- 64.09) was higher than that of group C (257.86 +/- 76.60). At T8, the anti-HBs level of group A (501.48 +/- 70.00) and group B (532.73 +/- 68.82) were higher than those of group C (256.12 +/- 75.39) (t =4.27, P = 0.0173).</p><p><b>CONCLUSION</b>Schemes of augmentation doses of Hepatitis B vaccine and being combined with GM-CSF should be in effect for non- and hypo-responders to Hepatitis B vaccine.</p>

Study on rhGM-CSF as adjuvant in revaccination among adults of non-and hyporesponders to hepatitis B vaccine / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the effect of recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) as adjuvant on immune response in adults of non-and hyporesponders to hepatitis B vaccine.</p><p><b>METHODS</b>Those who were once immunized with recombined yeast gene hepatitis B vaccine more than one standard scheme in two years and negative for hepatitis B markers were randomly sorted as group A and group B. 33 adults of group A were given hepatitis B vaccine 10 microg each time. The immune procedure was 0, 1 and 6 month. 34 adults of group B were given rhGM-CSF 300 microg for the first day, then 10 microg each time for routine immune. The blood samples were collected before the first injection and in 1, 2 and 8 months (T1, T2, T8) following the first injection to test Anti-HBs.</p><p><b>RESULTS</b>Anti-HBs positive conversion rates of group A and B at T8 was 39.39% and 64.71% respectively (P = 0.038). Anti-HBs levels of group B at T1, T2, T8 were (113.85 +/- 198.56) mIU/ml, (312.40 +/- 349.44) mIU/ml, (427.74 +/- 411.58) mIU/ml (P = 0.001). There was significant difference between group A and B in T8 Anti-HBs levels (P = 0.010).</p><p><b>CONCLUSION</b>Better immune response was found in the group of rhGM-CSF with hepatitis B vaccine. So rhGM-CSF can induce the immune respond to hepatitis B vaccine.</p>

Effect of shRNA-mediated silence of H-ras gene on proliferation of human SACC-M cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To examine the effects of H-ras gene silence on cell cycle, proliferation and apoptosis of salivary adenoid cystic carcinoma -M (SACC-M) cell lines.</p><p><b>METHODS</b>The plasmid H-ras-shRNA, containing the sequence of shRNA targeting H-ras, and HK-shRNA (without interfering effect) were constructed and transfected into SACC-M cells. The cell line with shRNA plasmid stable expression was isolated by G418. The expression levels of H-ras were detected by RT-PCR and protein immunofluorescent assay; cell cycle and cell apoptosis were analyzed by flow cytometry (FCM). The proliferation of cell was also determined by subcutaneous tumor formation in nude mice.</p><p><b>RESULTS</b>After transfection of H-ras-shRNA plasmid, the mRNA expression of H-ras in SACC-M cells was down-regulated by 61.80% and protein expression of H-ras was inhibited by 62.76%; the cell proliferation was inhibited obviously; the G0G1 phase cells were increased. The cell apoptosis rate of H-ras-shRNA group was significantly higher than that of HK-shRNA group (P <0.05). The volume of subcutaneous tumor in nude mice was significantly smaller in Hras-shRNA group than in control group.</p><p><b>CONCLUSIONS</b>The recombinant plasmid HRAS-shRNA could efficiently down-regulate the expression of H-ras gene and protein, induce apoptosis of SACC-M cells and simultaneously inhibit proliferation of these cells in vitro and in vivo.</p>

Correlation between DNA methylation of the ABO gene promoter CpG island and leukemia / 中国实验血液学杂志

Recent studies have found that ABO blood group antigen is also closely related to the onset and development of many diseases. More and more attention is being paid to the decrease of A/B blood group antigen caused by some tumors. This study was purpose to investigate the correlation between DNA methylation of the ABO gene promoter CpG island and leukemia. The relative contents of ABH antigen on the surface of RBC from kinds of blood disease patients and healthy individuals were detected by using flow cytometry and confocal laser scanning microscopy. The DNA sequences and CpG methylation of ABO gene promoter in patients with hematopathy and healthy individuals, as well as the -102 site methylation of ABO gene promoter in patients with hematopathy and healthy individuals were detected by PCR and MSP-PCR respectively. The results showed that RBC from leukemia patients displayed different degree of A/B antigen decrease. The sequences of ABO gene promotor of patients with hematopathy were not different from healthy individuals indicating high conservation of promoter sequences. Comparison of sequences between patients with hematopathy and healthy individual indicated that CpG islands on ABO gene promoter either from blood disease patients or from healthy individual had no methylated site in AA patients, but C residues at position -102, -101, -100, -99 and -97 on the promoter of ABO gene in AML, CML, ALL and some MDS patients were methylated. It is concluded that methylation of CpG islands in promoter of ABO gene may result in AB antigen decrease in patients with leukemia. The methylation sites -102, -101, -100, -99 and -97 may be specific for leukemia. The methylation of site -102 can be used as a molecular marker in differential diagnosis for leukemias.