OBJECTIVETo explore the clinical significance of HBV large protein (HBV-LP) in diagnosing viral replication, we detected the HBV-LP, HBV DNA and the hepatitis B viral markers (HBV M) in the serum of the patients infected with HBV.

METHODSHBV-LP and HBV M were analyzed by using ELISA. HBV-DNA was quantitatively detected using real-time fluorescent polymerase chain reaction.

RESULTS(1) No significant difference between the detectable rate of HBV DNA and HBV-LP was found in the same HBV M (P 0.05). (2) No significant difference between the positive rate of HBV DNA and HBV-LP was found in HBeAg negative patients. (3) The contents of serum HBV-LP was positively correlated with the number of HBV DNA copies.

CONCLUSIONThere was a close correlation between the positive rate of HBV-LP and HBV DNA, and HBV-LP is a reliable serological marker that can reflect the replication of HBV.

Adolescent ; Adult ; Aged ; DNA, Viral ; blood ; Female ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; physiology ; Humans ; Male ; Middle Aged ; Virus Replication

OBJECTIVETo investigate the expressions of chemokine receptors and interleukin (IL) receptors on the peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients and their correlations with clinical features as well as SLE disease activity index (SLEDAI).

METHODSThe mRNA expressions of chemokine receptors and IL receptors on PBMCs of 93 SLE patients and 30 healthy controls were detected by reverse transcription-polymerase chain reaction, including CCR2, CCR3, CCR4, CCR5, CCR6, CCR8, CXCR3, CXCRS, CX3CR1, XCR1, IL-4R, and IL-10R. The clinical features of SLE patients were recorded. The correlations of chemokine receptors and IL receptors mRNA expressions with clinical features as well as SLEDAI were assayed using linear regression analysis.

RESULTSThe level of CCR5 mRNA in SLE patients (including active and inactive SLE) was significantly higher than that in healthy controls (P < 0.05), and there was no significant difference between active and inactive patients in this respect (P > 0.05). CX3CR1 mRNA expression significantly increased from healthy control to inactive SLE to active SLE in sequence. The others (except for CCR8, CXCR3, and IL-10R) in active SLE patients were significantly higher than those in both inactive SLE patients and healthy controls (all P < 0.05). There were positive correlations between SLEDAI and CCR2 (r = 0.424, t = 4.313, P < 0.001), CCR3 (r = 0.518, t = 5.410, P < 0.001), CCR4 (r = 0.376, t = 3.851, P < 0.001), CCR6 (r = 0.457, t = 4.513, P < 0.001), CXCR5 (r = 0.455, t = 4.629, P < 0.001), CX3CR1 (r = 0.445, t = 4.523, P < 0.001), as well as XCR1 (r = 0.540, t = 5.445, P < 0.001). And CCR5 mRNA expression level was positively correlated with IL-4R mRNA (r = 0.313, t = 2.353, P < 0.05). The patients with myositis and cutaneous vasculitis simultaneously showed lower levels of CCR5 and CX3CR1, and CCR5 expression was negatively correlated with the scores of SLEDAI in SLE cases accompanied by photosensitivity (r = 0.426, t = -2.155, P < 0.05).

CONCLUSIONIncreased expressions of CCR5 and CX3CR1 on PBMCs may be indicators in clinical survey for SLE.

Adolescent ; Adult ; CX3C Chemokine Receptor 1 ; Child ; Female ; Humans ; Leukocytes, Mononuclear ; immunology ; Lupus Erythematosus, Systemic ; etiology ; immunology ; Male ; Middle Aged ; RNA, Messenger ; blood ; Receptors, CCR5 ; genetics ; Receptors, Chemokine ; genetics ; Receptors, Interleukin-10 ; genetics ; Receptors, Interleukin-4 ; genetics

OBJECTIVETo evaluate the impact of a new CD44 variant on invasion of human breast cancer cell line MCF-7, and its possible mechanisms.

METHODSThe full length cDNA encoding CD44v17 was obtained from the total RNA isolated from the MCF-7/ADR cells by reverse transcript-polymerase chain reaction (RT-PCR) and subcloned into pMD19-T vector. The CD44v17 gene sequence and reading frame were confirmed by two restriction enzymes and nucleotide sequencing, and then inserted into the eukaryotic expression vector pcDNA3.1. The pcDNA3.1-CD44v17 was transfected into MCF-7 cells by Lipofectamine. The changes of MMP-2 and MMP-9 expression at gene and protein levels were detected by RT-PCR and gelatin zymography, respectively. The number of the cells through the artificial matrix membrane in every group was counted to compare the change of the invasive ability regulated by CD44 variant. The ERK and p-ERK were investigated by Western blotting to approach the molecular mechanisms of MMP-2 and MMP-9 expression regulated by CD44 variant.

RESULTSThe new gene sequence was successfully cloned into recombinant vector pcDNA3.1 and identified by the two restriction enzymes. It was confirmed that the reconstructed plasmid contained the sequence of CD44 gene variant which was composed of 1 to 4 exons, 16 to 17 exons, and 1 to 205 bases of 18 exons. The new gene sequence was sent to NCBI for publication and obtained the registered number FJ216964. The up-regulated levels of the CD44 gene mRNA and protein were respectively detected by RT-PCR and flow cytometry in MCF-7 cells transfected with pcDNA3.1-CD44v17. The invasiveness of the cells and the activity of MMP-2 and MMP-9 were clearly activated by hyaluronic acid (HA) treatment and blocked by CD44 neutralizing antibody. Pretreated MCF-7/CD44v17 cells with the neutralizing antibody against CD44 and the inhibitor of MAPKs signaling pathway strongly block the expression of p-ERK.

CONCLUSIONA new CD44 gene variant has been found in adriamycin-resistant human breast cancer MCF-7/ADR cells. The expression vector pcDNA3.1-CD44v17 has been cloned and constructed successfully. HA can be integrated with CD44 variant and then regulates the expression of MMP-2 and MMP-9, which increases the invasion ability of MCF-7 cells through the Ras/MAPK signaling pathway.

Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Genetic Vectors ; Humans ; Hyaluronan Receptors ; genetics ; metabolism ; Hyaluronic Acid ; pharmacology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Neoplasm Invasiveness ; Phosphorylation ; Plasmids ; Protein Isoforms ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Signal Transduction ; Transfection

OBJECTIVETo study the effects of MTA1 small interfering RNA (siRNA) on the anchorage-independent growth and anoikis of prostate cancer cell line PC-3.

METHODSAfter transfection of human prostate cancer PC-3 cells by MTA1 siRNA, we detected the expression of the MTA1 gene by real-time PCR and Western blot, the anchorage-independent growth of the cells by clone formation in soft agar, and their anoikis by DNA fragmentation assay and flow cytometry.

RESULTSCompared with the control group, MTA1 siRNA transfection significantly decreased the mRNA and protein levels of MTA1, inhibited the anchorage-independent growth of the PC-3 cells, and induced their anoikis, all in a dose- and time-dependent manner (r = 0.935, P = 0.001; r = 0.901, P = 0.0005; r = 0.916, P = 0.0003).

CONCLUSIONMTA1 siRNA can inhibit the anchorage-independent growth of prostate cancer cells by inducing their anoikis.

Anoikis ; genetics ; Gene Expression Regulation, Neoplastic ; Histone Deacetylases ; genetics ; Humans ; Male ; Prostatic Neoplasms ; genetics ; RNA, Small Interfering ; Repressor Proteins ; genetics ; Transfection ; Tumor Cells, Cultured

Objective: To explore the effect of the comprehensive intervention on overweight and obesity among middle school students at the population level (health education lecture and official account push) and individual level (personalized dietary guidance), so as to provide a reference for preventing and controlling their overweight and obesity. Methods: Three junior high schools and three senior high schools were randomly selected in Guangzhou in 2018 by convenience sampling. Through physical examination, 1 457 overweight and obese students aged from 12 to 18 years old were screened. Intervention was administered through "Student Personalized Dietary Guidance" manual, health tweets on the official accounts, and health education lectures from September 2018 to December 2019. The Chi square test was used to compare the difference in overweight and obesity constituent ratio between the two groups before and after the intervention. And intervention effect was evaluated by analyzing the number needed to treated(NTT). Results: The proportion of overweight before the intervention was 66.71% (972/1 457), and decreased to 59.92% (873/1 457) after the intervention; the proportion of obesity before the intervention was 33.29% (485/1 457), which decreased to 26.63% (388/1 457) after the intervention. Among obese students, the smallest NNT was seen in the girl group aged 12-13 years (NNT=2.6, 95% CI =1.9-4.1), while the largest NNT in the boy group aged 14-18 years (NNT=5.9, 95% CI =4.7-8.1). The NNT of the girls aged 12-13 years was the smallest (NNT=2.7, 95% CI =2.2-3.5), and the NNT of the boys aged 14-18 years was the largest (NNT=7.4, 95% CI =6.0-9.7). Conclusion Health education at population level (health education lectures, official account push) with individual level (personalized dietary guidance) can effectively intervene overweight and obesity among middle school students in Guangzhou.

Objective: To investigate the distribution and drug resistance situation of staphylococcus aureus (SA) and methicillin-resistant staphylococcus aureus (MRSA) from the classroom environments in primary schools of Guangzhou. Methods: The air and the surfaces of door handles, desks, chairs, light switches and floor were sampled in the classrooms of 8 primary schools selected through stratified clustering method in Guangzhou from May to June, 2016. SA and MRSA were isolated and identified, and drug sensitivity tests were conducted. Results: A total of 760 samples were collected, the detection rate of SA and MRSA were 8.8% and 4.2%, respectively. There were statistically significant differences in the detection rate of staphylococcus aureus among different sampling sites(P<0.01).Detection of SA and MRSA on the floor,light’s witches and surface of deskes was both above 6.0%. The multiple drug resistance rate of MRSA was up to 100.0%, and the main resistance mode was Penicillin-Erythromycin-Rifampin-Tetracycline-Teicolanin. Conclusion MRSA can be detected in air, door handles, desk surface, chair surface, light switch and floor of primary schools. Relevant administration departments should pay attention to the environments health of Guangzhou primary schools.

Objective: To investigate the antibiotic resistance spectrum and genetic characteristics of multidrug-resistant Staphylococcus aureus(MDRSA) nasal isolate among primary school students, and to provide a scientific basis for the prevention and control of masal MDRSA resistance and the selection of clincal drugs in children. Methods: Antibiotic susceptibility experiments were performed on all SA isolates of 1 705 primary school students from 8 primary schools in Guangzhou selected by using multistage cluster stratified sampling method. MDRSA antibiotic susceptibility spectrum was analyzed, and the resistant, virulence and immune evasion cluster(IEC) genes detected by polymerase chain reaction(PCR). Results: The prevalence of MDRSA nasal carriage was 20.76%(354/1 705), and the proportion of multidrug resistance among SA isolates was 96.20%(354/368). The predominant resistant antibiotics of MDRSA isolates were penicillin(99.72%), erythromycin(96.33%), clindamycin(90.96%) and teicoplanin(90.11%). Notably, 240(67.80%, 240/354) MDRSA isolates were resistant to more than six antimicrobial categories. And the predominant detection rates of resistant genes were BlaZ(92.66%), Tet(M)(49.72%), virulence genes Tst(25.42%) and IEC genes Sak(92.09%), Hlb(61.58%). Conclusion We found high prevalence of nasal colonization MDRSA from healthy children. Moreover, MDRSA isolates has a high resistant rate to multiple antibiotics, and the proportion of resistant to ≥6 antimicrobial categories is high.

Objective To explore the role and mechanism of HMGB1 in the fatty acid metabolism reprogramming and mitochondrial fusion/fission of hypoxic and nutrient-poor pancreatic cancer cells. Methods The correlation between the expression level of HMGB1 in pancreatic cancer tissue and the survival rate of pancreatic cancer patients were analyzed by GEPIA database. CCK-8 assay was used to measure cell proliferation rate, and scratch test and Transwell chamber method were carried out to detect the effects of endogenous HMGB1 on the invasion and migration abilities of human pancreatic cancer cell line Patu8988 after hypoxic and nutrient-poor treatment. Laser confocal microscope was used to observe the changes of mitochondrial morphology of Patu8988 cells. Western blot was used to detect the expression levels of mitochondrial fusion/fission and de novo fatty acid synthesis-related proteins. Results GEPIA database analysis results showed that HMGB1 was highly expressed in pancreatic cancer tissues (P < 0.01), and the expression level was negatively correlated with the survival time of pancreatic cancer patients (P=0.00097). Knockdown of HMGB1 expression could inhibit the proliferation, invasion and migration abilities of Patu8988 cells under hypoxic and nutrient-poor conditions. However, mitochondrial fission in patu8988 cells was increased. Knockdown of HMGB1 in Patu8988 cells increased the expression of fission-related protein FIS1 while decreased the expression of p-DRP1(Ser637) and fusion-related protein MFN1 and MFN2 in hypoxic and nutrient-poor environment; ACLY, p-ACLY and FASN protein expression levels were down-regulated. Conclusion Endogenous HMGB1 can promote the fusion and inhibit the fission of mitochondria in hypoxic and nutrient-poor Patu8988 cells, maintain mitochondrial morphology and function, and thereby up-regulate ACLY protein expression and phosphorylation level, promote FA synthesis, and maintain the proliferation, invasion and migration abilities of pancreatic cancer cells.

Objective: To explore the association between maternal pre pregnancy and pre delivery overweight with overweight and obesity among offspring during adolescence in Guangzhou, and to provide evidence for child obesity prevention. Methods: Based on the routine physical examination of primary and secondary school students in Guangzhou, random sampling was used to 6 middle schools and questionnaire survey was conducted among 3 384 students and their parents. Students with overweight and obesity were included in the case group, and the other students were included in the control group. Propensity Score Matching (PSM) was adopted to reduce selection bias. Logistic regression model and χ 2 test were used to analyze the data before and after PSM. Results: The result of univariate analysis showed that there were statistically significant differences between overweight/obese group and the control group by gender, schooling stage (middle and high schools), picky eater, family history of obesity, family monthly income, delivery mode, high birthweight, and gestational weight gain before PSM( χ 2=42.38, 10.64, 14.47, 26.85, 10.58, 13.59 , 15.53, 20.64, P <0.05). After PSM, results showed that there were no statistically significant differences between overweight/obese group and the control group in middle and high schools, and mother delivery mode( P >0.05). Logistic regression analysis showed that the risk of overweight and obesity of maternal pre pregnancy on adolescent offspring was 1.54 times higher than control group (95% CI =1.01-2.36) before PSM, and the overweight and obesity of maternal predelivery also increased the risk of overweight and obesity of adolescent offspring( OR=2.35, 95%CI =1.67-3.31). After PSM, maternal overweight and obesity pre pregnancy ( OR=2.17, 95%CI =1.41-3.34) and maternal overweight and obesity pre delivery( OR=2.99, 95%CI =2.08-4.31) significantly increased the risk of overweight and obesity in adolescent offspring. Conclusion Maternal overweight and obesity pre pregnancy and pre delivery are associated with increased risk of overweight and obesity in adolescent offspring.

Objective To explore the ability of computed?tomography ( CT) radiomic features to predict the Epidermal growth factor receptor ( EGFR ) mutation status and the therapeutic response of advanced lung adenocarcinoma to EGFR? Tyrosine kinase inhibitors ( TKIs ) treatment. Methods A retrospective analysis was performed on 253 patients diagnosed as advanced lung adenocarcinoma, who underwent EGFR mutation detection, and those with EGFR sensitive mutation were treated with TKIs. Using the Lasso regression model and the 10 fold cross?validation method, the radiomic features of predicted EGFR mutation status and the screening of TKIs for sensitive populations were obtained.715 radiomic features were extracted from unenhanced, arterial phase and venous phase, respectively. Results The area under curve (AUC) values of the multi?phases including unenhanced, arterial phase and venous phase of the EGFR mutation status validation group were 0.763, 0.807 and 0.808, respectively. The number of radiomic features extracted from the multi?phases were 5, 18 and 23, respectively, which could distinguish the EGFR mutation status. The AUC values of the multi?phases of the EGFR?TKIs sensitive validation group were 0.730, 0.833 and 0.895, respectively. The number of radiomic features extracted from the multi?phases were 3, 7 and 22, respectively, which can be used to screen the superior population for TKIs treatment. The efficiency of radiomic features extracted from venous phase in predicting EGFR mutant status and EGFR?TKIs sensitivity was significantly superior than those of unenhanced and arterial phase. Conclusions The radiomic features of CT scanning can be used as the radiomics biomarker to predict the EGFR mutation status of lung adenocarcinoma and to further screen the dominant population in TKIs therapy, which provides the basis for targeted therapy.