Transition rules of cognitive frailty and influencing factors in the elderly in China / 中华流行病学杂志

Objective: To understand the transition rules of cognitive frailty and its influencing factors in the elderly in China and provide evidence for the early intervention of cognitive frailty. Methods: Data were retrospectively collected from China Health and Retirement Longitudinal Study with 3 round consecutive survey (2011, 2013, 2015) and the state of the subjects were classified into four categories: robust-normal cognitive, cognitive impairment, physical frailty, and cognitive frailty. A multi-state Markov model was established to explore the transition rules of cognitive frailty and its influencing factors. Results: A total of 3 470 older adults were included, and 350 (10.09%) had cognitive frailty at baseline. After two years, the probability of cognitive frailty in the cognitive impairment population was higher than that in people with physical frailty (31.6% vs. 7.6%). Persons with cognitive frailty were more likely to become physical frailty (29.7% vs. 15.6%). Being women (HR=1.599, 95%CI: 1.058-2.417), comorbidity (HR=3.035, 95%CI: 1.090-8.450), and depression (HR=1.678, 95%CI: 1.153-2.441) were the risk factors associated with cognitive frailty in the elderly, while being educated (HR=2.367, 95%CI: 1.567-3.575) was a protective factor for the transition of cognitive frailty to physical frailty. Conclusions: The prevalence of cognitive frailty is relatively high in the elderly in China. Those with cognitive impairment have a higher probability of cognitive frailty. Gender, education level, comorbidity, and depression are the main influencing factors for the occurrence and transition of cognitive frailty.

Family history of esophageal cancer modifies the association of serum lipids and malignant esophageal lesions: a nested case-control study from the "Endoscopic Screening for Esophageal Cancer in China" trial / 中华医学杂志(英文版)

BACKGROUND@#The association of lipids and cancer has varied greatly among different cancer types, lipid components and study populations. This study is aimed to investigate the association of serum lipids and the risk of malignant lesions in esophageal squamous epithelium.@*METHODS@#In the "Endoscopic Screening for Esophageal Cancer in China" (ESECC) trial, serum samples were collected and tested for total cholesterol (TC), triglycerides, low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol at the time of subject enrollment. Cases were defined as malignant esophageal lesions identified by baseline endoscopic examination or by follow-up to May 31, 2018. Controls were randomly selected using incidence density sampling in the same cohort. Conditional logistic models were applied to identify the association of serum lipids and the risk of malignant esophageal lesions. Effect modification was evaluated by testing interaction terms of the factor under assessment and these serum lipid indicators.@*RESULTS@#No consistent association between serum lipid levels and esophageal malignant lesions were found in a pooled analysis of 211 cases and 2101 controls. For individuals with a family history of esophageal cancer (EC), high TC, and LDL-C were associated with a significantly increased risk of having malignant lesions (odds ratio [OR]High vs. Low TC = 2.22, 95% confidence interval [CI]: 1.14-4.35; ORHigh vs. Low LDL-C = 1.93, 95% CI: 1.01-3.65). However, a negative association was observed in participants without an EC family history (ORHigh vs. Low TC = 0.69, 95% CI: 0.48-0.98, Pinteraction = 0.002; ORHigh vs. Low LDL-C = 0.50, 95% CI: 0.34-0.76, Pinteraction < 0.001).@*CONCLUSIONS@#In this study, we found that the association of serum lipids and malignant esophageal lesions might be modified by EC family history. The stratified analysis would be crucial for population-based studies investigating the association of serum lipids and cancer. The mechanism by which a family history of EC modifies this association warrants further investigation.

Clinical features of 109 cases of novel coronavirus pneumonia / 中华传染病杂志

Objective@#To observe theepidemiology, clinical manifestations, laboratory tests, imaging findings, treatment and prognosis of patients with novel coronavirus pneumonia.@*Methods@#Clinical data of 109 patients with suspected and definite novel coronavirus pneumonia admitted to Wuhan Sixth Hospital from December 24, 2019 to January 28, 2020 were retrospectively analyzed. Statistical analysiswas performed by using t test or chi-square test.@*Results@#Among the 109 patients, 48 (44%) were male and 61 (56%) were female, with the average age of (52.5±10.8) years. Fifty-four patients (49.5%) had definite contact history. Among the 109 patients, 104 (95.5%) presentedwith fever, 37(33.7%) with headache, 78 (71.9%) with general pain, 88 (80.8%) with fatigue and poor appetite, 23 (21.3%) with diarrhea, 94 (86.5%) withcoughing, 23 (21.3%) with shortness of breath, 57 (52.8%) withpalpitation, 45 (41.5%) with chest distress, 4 (3.3%) with chest pain, 40 (37.0%) with lung rales. Forty-two cases (38.5%) had leukocyte count <4×109/L, 58 cases (53.2%) had lymphocyte count <1.5×109/L, 7 cases (24.8%) had hemoglobin <120g/L, 37 cases(33.9%) had LDH >230 mmol/L, 29 cases (26.6%) had brain natriuretic peptide precursor>300 ng/mL, 87 cases (79.8%) had hypersensitive C-reactive protein >10mg/L, 26 cases (23.9%) had D-dimer >0.5 mg/L, 35 cases (32.1%) had coagulation disorder. The leukocyte counts, LDH, brain natriuretic peptide precursor and D-dimer of severe/critical cases[(11.33±4.87)×109/L, (527.51±260.87) mmol/L, (722.88±189.56) ng/mL, (1.89±4.24) mg/L, respectively] were all significantly higher than those of common cases [(4.02±1.49)×109/L, (159.75±30.31)mmol/L, (428.22±124.76)ng/mLand (0.41±0.22)mg/L, respectively], while the lymphocyte count of severe/critical cases [(0.60±0.17)×109/L] was significantly lower than common cases [(1.13±0.43)×109/L] (t=11.36, 11.33,9.81,2.81 and 7.77,all P<0.05). On admission, chest CT showed that 27 cases (24.8%) of pneumonia were unilateral, 82 cases (75.2%) werebilateral, and most of them were ground glass. The pneumonia progressed in a short time and reached the peak within 10 days. The comprehensive treatment included antiviral drugs, prevention ofbacterialinfection and supportive treatment, and glucocorticoid and respiratory support treatment wereadministrated when necessary.@*Conclusions@#The novel coronavirus pneumonia is characterized by highly infectious, rapid progress, and diverse clinical and imaging features. Early diagnosis and active comprehensive treatment could improve theprognosis and reduce themortality.

Individual Identification of Cartilage by Direct Amplification in Mass Disasters / 法医学杂志

OBJECTIVES@#To explore the effectiveness of direct amplification for the STR analysis of cartilage, and to accelerate the effectiveness of disaster victim identification.@*METHODS@#Eighty-eight cartilage samples were directly amplified by PowerPle® 21 kit, and the results of genotyping were compared with that obtained by the magnetic beads method.@*RESULTS@#In 88 cartilage samples, the STR genotypes were successfully detected from 84 samples by direct amplification and magnetic beads method, and both the results of genotyping by two method were consistent.@*CONCLUSIONS@#Direct amplification with PowerPlex® 21 kit can be used for STR genotyping of cartilages. This method is operated easily and promptly, which has a potential application in the individual identification of mass disasters.

Effects of BCYRN1 on miR-503 on migration and invasion of lung cancer by Notch1 pathway / 中国免疫学杂志

Objective:To investigate the mechanism of BCYRN1 regulating miR-503 through Notch1 signaling pathway in the migration and invasion of lung cancer. Methods:The expression of BCYRN1 and miR-503 in different lung cancer cell lines were detected by qPCR. Immunofluorescence and qPCR were used to detect the transfection efficiency of lentivirus BCYRN1 + siRNA transfected lung cancer cells. Double luciferase reporter gene was used to detect the interaction between BCYRN1 and miR-503. Transwell invasion test and scratch test were used to detect the invasion and migration of lung cancer cells after silencing BCYRN1. Western blot was used to detect the expression of Notch1 signaling pathway silence BCYRN1. The effect of silencing BCYRN1 on lung cancer cells in nude mice was measured by subcutaneous tumor formation in nude mice. Results:The expression level of BCYRN1 was the highest in lung cancer cell H1299,and the expression level of miR-503 was relatively high. Immunofluorescence and mRNA levels demonstrated that BCYRN1 + siRNA lentivirus could be effectively transfected into H1299 cells;BCYRN19 binds specifically to the 3′-UTR of miR-503;silencing BCYRN1 could inhibit the invasion and migration of lung cancer H1299 cells;the expression of Notch1 pathway protein was down-regulated after silencing BCYRN1. Compared with NC group,tumor volume and weight of BCYRN1-siRNA group were significantly decreased. Conclusion:BCYRN1 can target the regulation of miR-503 through Notch1 signaling pathway in the invasion and migration of lung cancer H1299 cells.

Effects of BCYRN1 on miR-503 on migration and invasion of lung cancer by Notch1 pathway / 中国免疫学杂志

Objective:To investigate the mechanism of BCYRN1 regulating miR-503 through Notch1 signaling pathway in the migration and invasion of lung cancer. Methods:The expression of BCYRN1 and miR-503 in different lung cancer cell lines were detected by qPCR. Immunofluorescence and qPCR were used to detect the transfection efficiency of lentivirus BCYRN1 + siRNA transfected lung cancer cells. Double luciferase reporter gene was used to detect the interaction between BCYRN1 and miR-503. Transwell invasion test and scratch test were used to detect the invasion and migration of lung cancer cells after silencing BCYRN1. Western blot was used to detect the expression of Notch1 signaling pathway silence BCYRN1. The effect of silencing BCYRN1 on lung cancer cells in nude mice was measured by subcutaneous tumor formation in nude mice. Results:The expression level of BCYRN1 was the highest in lung cancer cell H1299,and the expression level of miR-503 was relatively high. Immunofluorescence and mRNA levels demonstrated that BCYRN1 + siRNA lentivirus could be effectively transfected into H1299 cells;BCYRN19 binds specifically to the 3′-UTR of miR-503;silencing BCYRN1 could inhibit the invasion and migration of lung cancer H1299 cells;the expression of Notch1 pathway protein was down-regulated after silencing BCYRN1. Compared with NC group,tumor volume and weight of BCYRN1-siRNA group were significantly decreased. Conclusion:BCYRN1 can target the regulation of miR-503 through Notch1 signaling pathway in the invasion and migration of lung cancer H1299 cells.

Determination of Related Substances in Dienestrol Tablets by HPLC / 中国药师

Objective:To determine the related substances in dienestrol tablets by HPLC. Methods:The separation was performed on a C18 column(250 mm × 4. 6 mm,5μm). The mobile phase consisted of methanol-acetonitrile(76∶24), the flow rate was set at 0. 8 ml·min-1 with the detection wavelength at 229 nm, and the sample size was 10 μl. Results:Under the established chromatographic conditions, the related substances and dienestrol were separated completely. The limit of detection was 2 ng. Conclusion:The method is simple, accurate, specific and sensitive in the determination of related substances in dienestrol tablets.

Semen quality of the men from six districts in Chongqing / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the quality and spatial distribution features of semen and to evaluate the reproductive health of the males in the Chongqing section of the Three-Gorge Reservoir area.</p><p><b>METHODS</b>We collected semen samples by masturbation after 2 -7 days of abstinence from the men in Nan'an, Shapingba, Zhongxian, Wanzhou, Yunyang and Wushan of Chongqing, which are geographically and demographically representative of the Three-Gorge Reservoir area. We analyzed the semen quality of all the samples and evaluated the reproductive health of the men.</p><p><b>RESULTS</b>The mean value of the five semen parameters of the male subjects from the six districts was within the normal range, including semen volume, sperm concentration, total sperm count, rapid progressive motile sperm, and total motile sperm. Those from Shapingba, Yunyang and Zhongxian exhibited abnormal sperm motility. According to the WHO criteria, normal value of all the semen parameters was found in less than 50% of the semen samples from the six districts, in 47% of those from Yunyang, and only 16% of those from Wanzhou. Spatial distribution maps of the semen parameters revealed significant spatial differences in seminal quality among the six districts, the highest in Yunyang, and the lowest in Wanzhou and Wushan that are located in the middle and lower reaches of the Three-Gorge Reservoir area.</p><p><b>CONCLUSION</b>The mean value of semen parameters was low in a large proportion of men in the Chongqing section of the Three-Gorge Reservoir area, with spatial differences along the Changjiang river.</p>

Expression of estrogen receptor α in hepatitis B-related hepatocellular carcinoma and its clinical significance / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the expression and its clinical significance of estrogen receptor (ERα) and phosphorylated estrogen receptor (p-ERα) in patients with hepatocellular carcinoma. The associations between ERα, p-ERα and IL-6 were also analyzed.</p><p><b>METHODS</b>Immunohistochemistry was used to detect the expression of ERα, p-ERα and IL-6 in tumor tissues from 77 cases with hepatocellular carcinoma. The relations between ERα and the clinical pathological parameters and prognosis were also analyzed.</p><p><b>RESULTS</b>The positive rates of ERα, p-ERα and IL-6 in hepatocellular carcinoma were 39.0% (30/77), 45.4% (35/77) and 72.7% (56/77), respectively. The expression of ERα and p-ERα were negatively correlated with the expression of IL-6 (r=-0.468, P<0.01; r=-0.370, P<0.01, respectively). The positive rate of ERα in patients with tumor size≤5 cm, serum level of alpha-fetoprotein<400 µg/L, with complete encapsulation and non-microvascular invasion was significantly higher than those with tumor size>5 cm, serum level of alpha-fetoprotein≥400 µg/L, non-complete encapsulation and with microvascular invasion (all P<0.05). The overall survival rates of ERα-positive and ERα-negative patients were 66.7% and 23.4% (P<0.05). And the disease-free survival rates of ERα-positive and ERα-negative patients were 83.3% and 57.4% (P<0.05).</p><p><b>CONCLUSIONS</b>The tumor biological features of ERα-positive patients are better than that of ERα-negative patients. The role of ERα in hepatocellular carcinoma may be related to IL-6 level.</p>

Influence of nm23-H1 gene silence in K562 cell on its differentiation toward megakaryocyte / 中华血液学杂志

<p><b>OBJECTIVE</b>To construct a stable nm23-H1-knock-down cell model with K562 cell line and study its differentiation toward megakaryocyte.</p><p><b>METHODS</b>Eukaryotic expression vector pSilencer 4.1-CMV-sinm23 expressing siRNA targeting nm23-H1 was transfected into K562 cells with lipofectamine2000. Cells with stably nm23-H1 silence were screened out by G418. Real-time quantitative PCR, immunocytochemistry, western blot were used to confirm the nm23-H1-knock-down K562 model. Cell differentiation capacity was detected by NBT reduction assay. Surface antigen Gp IIb-IIIa (CD41) of knock-down cells treated with phorbol 12-myristate 13-acetate was analyzed by flow cytometry. Western blot was used to detect the ERK1/2 signal pathway after the stimulation of phorbol 12-myristate 13-acetate.</p><p><b>RESULTS</b>Endogenous nm23-H1 was silenced by pSilencer 4.1-CMV-sinm23 and the silence efficiency was up to 75% and 70% in mRNA and protein levels respectively compared with the mock cells. Under phorbol 12-myristate 13-acetate treatment, the knock-down cells displayed a significantly increased differentiation ability toward megakaryocyte compared with control. The NBT reduction values were (0.31 +/- 0.07) and (0.23 +/- 0.05) respectively. Further results revealed that nm23-H1 gene regulating the megakaryocytic differentiation was due in part to the increased ERK1/2 phosphorylation.</p><p><b>CONCLUSIONS</b>A stable nm23-H1-knock-down K562 cell model is successfully constructed. nm23-H1 involves in regulating the megakaryocytic differentiation of K562 cell line.</p>

Immune Responses and Histopathological Changes in Rabbits Immunized with Inactivated SARS Coronavirus / 中国病毒学

To evaluate the immunogenicity of inactivated SARS coronavirus (SARS-CoV), three groups of rabbits were immunized three times at 2-week intervals with inactivated vaccine + adjuvant, adjuvant,and normal saline respectively. Eight batchs of serum were sampled from the auricular vein at day 7 to day 51, and specific IgG antibody titers and neutralizing antibody titers were detected by indirect ELISA and micro-cytopathic effect neutralizing test. Antibody specificity was identified by proteinchip assay.Histopathological changes were detected by H&E staining. The results showed that, rabbits in the experimental group immunized with inactivated SARS-CoV all generated specific IgG antibodies with neutralizing activity, which suggested the inactivated SARS-CoV could preserve its antigenicity well and elicit an effective humoral immune responses. The peak titer value of specific IgG antibody and neutralizing antibody reached 1:40960 and 1:2560 respectively. In the experimental group, no obvious histopathological changes was detected in the H&E stained slides of heart, spleen, kidney and testis samples, but the livers had slight histopathological changes, and the lungs presented remarkable histopathological changes. These findings are of importance for SARS-CoV inactivated vaccine development.

Variation analysis of the severe acute respiratory syndrome coronavirus putative non-structural protein 2 gene and construction of three-dimensional model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The rapid transmission and high mortality rate made severe acute respiratory syndrome (SARS) a global threat for which no efficacious therapy is available now. Without sufficient knowledge about the SARS coronavirus (SARS-CoV), it is impossible to define the candidate for the anti-SARS targets. The putative non-structural protein 2 (nsp2) (3CL(pro), following the nomenclature by Gao et al, also known as nsp5 in Snidjer et al) of SARS-CoV plays an important role in viral transcription and replication, and is an attractive target for anti-SARS drug development, so we carried on this study to have an insight into putative polymerase nsp2 of SARS-CoV Guangdong (GD) strain.</p><p><b>METHODS</b>The SARS-CoV strain was isolated from a SARS patient in Guangdong, China, and cultured in Vero E6 cells. The nsp2 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into eukaryotic expression vector pCI-neo (pCI-neo/nsp2). Then the recombinant eukaryotic expression vector pCI-neo/nsp2 was transfected into COS-7 cells using lipofectin reagent to express the nsp2 protein. The expressive protein of SARS-CoV nsp2 was analyzed by 7% sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The nucleotide sequence and protein sequence of GD nsp2 were compared with that of other SARS-CoV strains by nucleotide-nucleotide basic local alignment search tool (BLASTN) and protein-protein basic local alignment search tool (BLASTP) to investigate its variance trend during the transmission. The secondary structure of GD strain and that of other strains were predicted by Garnier-Osguthorpe-Robson (GOR) Secondary Structure Prediction. Three-dimensional-PSSM Protein Fold Recognition (Threading) Server was employed to construct the three-dimensional model of the nsp2 protein.</p><p><b>RESULTS</b>The putative polymerase nsp2 gene of GD strain was amplified by RT-PCR. The eukaryotic expression vector (pCI-neo/nsp2) was constructed and expressed the protein in COS-7 cells successfully. The result of sequencing and sequence comparison with other SARS-CoV strains showed that nsp2 gene was relatively conservative during the transmission and total five base sites mutated in about 100 strains investigated, three of which in the early and middle phases caused synonymous mutation, and another two base sites variation in the late phase resulted in the amino acid substitutions and secondary structure changes. The three-dimensional structure of the nsp2 protein was successfully constructed.</p><p><b>CONCLUSIONS</b>The results suggest that polymerase nsp2 is relatively stable during the phase of epidemic. The amino acid and secondary structure change may be important for viral infection. The fact that majority of single nucleotide variations (SNVs) are predicted to cause synonymous, as well as the result of low mutation rate of nsp2 gene in the epidemic variations, indicates that the nsp2 is conservative and could be a target for anti-SARS drugs. The three-dimensional structure result indicates that the nsp2 protein of GD strain is high homologous with 3CL(pro) of SARS-CoV urbani strain, 3CL(pro) of transmissible gastroenteritis virus and 3CL(pro) of human coronavirus 229E strain, which further suggests that nsp2 protein of GD strain possesses the activity of 3CL(pro).</p>

Humoral immune responses in rabbits induced by an experimental inactivated severe acute respiratory syndrome coronavirus vaccine prepared from F69 strain / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The etiologic agent of severe acute respiratory syndrome (SARS) has been confirmed to be a novel coronavirus (CoV), namely SARS-CoV. Developing safe and effective SARS-CoV vaccines is essential for us to prevent the possible reemergence of its epidemic. Previous experiences indicate that inactivated vaccine is conventional and more hopeful to be successfully developed. Immunogenicity evaluation of an experimental inactivated SARS-CoV vaccine in rabbits was conducted and reported in this paper.</p><p><b>METHODS</b>The large-scale cultured SARS-CoV F69 strain was inactivated with 0.4% formaldehyde and purified, then used as the immunogen combined with Freund's adjuvant. Eight adult New Zealand rabbits were immunized four times with this experimental inactivated vaccine. Twelve sets of rabbit serum were sampled from the third day to the seventy-fourth day after the first vaccination. The titers of specific anti-SARS-CoV IgG antibody were determined by indirect enzyme-linked immunosorbent assay, and the neutralizing antibody titers were detected with micro-cytopathic effect neutralization test.</p><p><b>RESULTS</b>Rapid and potent humoral immune responses were induced by the inactivated SARS-CoV vaccine in all the eight test rabbits. Titers of both specific IgG antibody and neutralizing antibody peaked at about six weeks after first vaccination, with the maximum value of 1:81 920 and 1:20 480, respectively. After that, serum antibody levels remained at a plateau or had a slight decrease, though two boosters were given in the succedent 4 to 5 weeks. Cross neutralization response existed between SARS-CoV F69 strain and Z2-Y3 strain.</p><p><b>CONCLUSIONS</b>The inactivated SARS-CoV vaccine made from F69 strain owns strong immunogenicity, and the cross neutralization response between the two different SARS-CoV strains gives a hint of the similar neutralizing epitopes, which provide stable bases for the development of inactivated SARS-CoV vaccines.</p>