In medical clinical study, the researchers and the ethics committee members' personal economic in-terests and responsibilities conflicts may produce damage to experimental research and the subjects, may make the study being questioned the authenticity and the objectivity of clinical trial results, also damage to the subjects' rights and interests and damage the credibility of hospital. In order to prevent the happening of the conflict, put forward the strategies:strengthen legislation construction, improve the treatment method, set up a conflict of interest com-mittee, adhere to the principle of open, review and restriction to the personnel, regularly organize researchers and ethics committee members attend the training.

Professional postgraduates in geriatric medicine are one of the most important re-courses of geriatric specialists. However, training model for geriatric postgraduates is still unclear in China. In general, geriatric medicine has no independent training plan, and is included in internal medicine training. Such model cannot represent the feature of geriatric medicine. The present paper aimed to discuss the ability construction for geriatric postgraduates based on the geriatric medicine specialist training experience both at home and abroad, including diagnosis and treatment of multi-system diseases, ability of geriatric assessment, management of geriatric syndromes, consultation for geriatric rehabilitation and ability for continuity health service in order to cultivate the compound tal-ents suitable for the aging society requirement for geriatric medicine in China.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) or high glucose on the toll-like receptor 4 (TLR4) expression,inflammatory cytokines and fibrotic factors in human tubular epithelial cells (HK-2),revealing the innate immune-related pathogenesis of diabetic nephropathy (DN) which may have clinical implications.Methods Three TLR4 siRNA sequences were designed and synthetized.After transfection,the most effective siRNA was selected to use for further expriments.The experiment consisted of 2 parts.Part 1:Cells were divided into three groups:normal-glucose group (NG,5.5mmol/L glucose),mannose group (M,5.5 mmol/L glucose + 19.5 mmol/L mannose),High-glucose group (HG,25 mmol/L glucose),preliminary validated the effects of high glucose and high osmotic pressure.Part 2:Cells were divided into seven groups:NG group,HG group,Ang Ⅱ group,Ang Ⅱ + negative group,HG+ negative group,Ang Ⅱ + siRNA group and HG+ siRNA group.Real time PCR was used to analyze the mRNA expression of TLR4,myeloid differentiation factor 88 (MyD88),heat shock protein 47 (HSP47).Western blotting was used to observe the protein expression of TLR4,MyD88,HSP47,NF-κB,type Ⅳ collagen (ColⅣ).ELISA was used to detect the expression of monocyte chemotactic protein-1 (MCP-1) and interleukin-6 (IL-6).Results Compared with NG group,TLR4,MyD88,HSP47 mRNA and TLR4,MyD88,NF-κB,ColⅣ,HSP47 protein were highly expressed under high glucose or Ang Ⅱconditions (P ＜ 0.01),and the expression levels of MCP-1 and IL-6 also increased significantly (P ＜ 0.01).Compared with HG or Ang Ⅱ group,the above indicators were obviously inhibited in the TLR4 siRNA groups (P＜0.01).Comparison between blank vector transfected groups and HG group as well as Ang Ⅱ group indicated no statistic significance (P ＞ 0.05).Conclusions Both Ang Ⅱ and high glucose stimulate TLR4 expression,which result in the up-regulation of inflammatory and fibrotic factors in HK-2.Specific target of TLR4 gene silencing can block the TLR4 pathway that is activated by high glucose and Ang Ⅱ,and thus reduce the inflammatory and fibtogenic factors' release.TLR4 signal is the common innate immune response pathway which induces the release of inflammatory and fibrotic factors in HK-2 under high glucose or high angiotension conditions.

OBJECTIVESince glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most cases have single nucleotide mutations in the coding region, and current test methods for gene mutation have some missed detections, this study aimed to investigate the feasibility of RT-PCR sequencing in the detection of gene mutation in G6PD deficiency.

METHODSAccording to the G6PD/6GPD ratio, 195 children with anemia of unknown cause or who underwent physical examination between August 2013 and July 2014 were classified into G6PD-deficiency group with 130 children (G6PD/6GPD ratio <1.00) and control group with 65 children (G6PD/6GPD ratio≥1.00). The primer design and PCR amplification conditions were optimized, and RT-PCR sequencing was used to analyze the complete coding sequence and verify the genomic DNA sequence in the two groups.

RESULTSIn the G6PD-deficiency group, the detection rate of gene mutation was 100% and 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 boys; 13 heterozygous missense mutations, 1 homozygous same-sense mutation (C1191T) which had not been reported in China and abroad, and 14 single nucleotide polymorphisms of C1311T were detected in 37 girls. The control group showed a high rate of missed detection of G6PD deficiency (carriers) in the specimens from girls (35%, 13/37).

CONCLUSIONSRT-PCR sequencing has a high detection rate of G6PD gene mutation and a certain value in clinical diagnosis of G6PD deficiency.

Adolescent ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Infant ; Male ; Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Objective To investigate the expression of miR-520d-3p in 2 and 4 Gy X-ray irradiated A549 cells, serum samples of non-small cell lung cancer (NSCLC) patients and its significance. Methods Real time quantitative PCR (RT-qPCR) was employed to detect the expression miR-520d-3p in 2 and 4 Gy X-ray irradiated A549 cells in vitro. Besides, 0, 2 and 4 Gy X-ray irradiated A549 cells were used to prepare animal model of nude mice lung metastasis through mainline. The expressions of miR-520d-3p and lung tissue of nude mice were detected, and the serum samples of NSCLC patients were collected to test the expression of miR-520d-3p . Results Compared with the control group ( 0 Gy ) , the expression of miR-520d-3p was up-regulated significantly in 2 and 4 Gy X-ray irradiated A549 cells after 48 hours (1.00±0.03, 1.47±0.10, 1.84 ±0.09 respectively), and there was a significant difference (F= 94.350, P= 0.000). Furthermore, compared the control group with injection after 10 weeks, the expressions of miR-520d-3p in nude mice lung tissue in 0, 2 and 4 Gy X-ray irradiated groups were increased (2.33 ±0.13, 8.24 ±0.25, 3.46 ±0.14, respectively) (F=1.787, P= 0.227), and the expressions in serum were increased (11.43±2.30, 10.22±4.62, 8.99 ±3.67, respectively) (F= 1.547, P= 0.246). Out of 20 serum samples of NSCLC patients (including 10 cases of adenocarcinoma, 10 cases of squamous carcinoma ), 11 cases (55%) were detected with up-regulated miR-520d-3p expression. Conclusions 2 and 4 Gy X-ray could increase the expression of miR-520d-3p of A549 cells in vitro and in vivo, which might be correlated with the enhanced invasion and metastasis of A549 cells induced by X-ray in vitro and in vivo. Furthermore, the expressions of miR-520d-3p were increased significantly in over 50%serum samples of NSCLC patients, which might be a new marker for the diagnosis of NSCLC.

Objective To examine the effects of benazepril and losartan on glomerular podocyte autophagy in aged spontaneously hypertensive rats (SHRs) and investigate the underlying mechanisms of renal-protective effects of angiotensin-converting enzyme inhibitors (ACEI)/angiotensin receptor blockers (ARB).Methods Wistar-Kyoto rats (WKYs) were used as the normal control (NORM) group (2 ml physiological saline per day).SHRs were randomly divided into 4 groups:the CTRL group (2 ml physiological saline per day),the ACEI group (10 mg · kg-1 · d-1),the ARB group (30 mg· kg-1 ·d-1) and the combined group (10 mg· kg-1 · d-1 benazepril and 30 mg· kg-1 · d-1),with six 18-month-old make rats in each group.The experiments were conducted during a 4-month period.Blood pressure was monitored regularly.At the end of the experiments,we measured the levels of urine protein,urine creatinine,serum creatinine (SCR),blood urea nitrogen (BUN),and serum and renal cortex angiotensin Ⅱ (AngⅡ).Ultrastructural changes in the kidney were examined under light and transmission electron microscopy.The expressions of nephrin,LC3BⅡ,Atg5 and p62 in the glomerulus were analyzed by Western blot analysis.Results After treatment,the blood pressure and the urine albumin/creatinine ratio of the four SHR groups were still significantly higher than those of the NORM group,but the blood pressure and the urine albumin/creatinine ratio of the ARB group and the combined group were significantly lower than those of the CTRL group (all P＜ 0.05);There were no significant differences in SCR and BUN levels among these five groups (P＞ 0.05);The level of serum AngⅡ of the combined group was significantly higher than that of the CTRL group [CTRL (0.08±0.00) μg/L,Combined (0.12±0.01) μg/L,P＜0.05];The levels of cortex AngⅡ of the four SHR groups were significantly lower than those of the NORM group,while the level of cortex AngⅡ of the ARB group was significantly higher than that of the CTRL group (all P＜0.05);Renal ultrastructural examination revealed shrunken glomeruli,fused or effaced epithelial cell foot processes,and focal atrophy of renal tubules in the four SHR groups.These pathological changes were more serious in the CTRL group but less so in the combined group.There were significantly more autophagosomes in the NORM group and the combined group than in the CTRL group (P＜0.05).Compared with the NORM group,the expressions of nephrin,LC3BⅡ,Atg5 and p62 in the CTRL group were suppressed significantly (P ＜ 0.05).The expressions of nephrin,LC3BⅡ and Atg5 in the ACEI group and the expressions of nephrin,LC3BⅡ,Atg5 and p62 in the ARB group and the combined group were higher than in the CTRL group (P＜0.05).Conclusions ACEI/ARB can decrease the autophagic activity of glomerular podocytes.The renal-protective effects of ACEI/ARB may be mediated by glomerular podocyte autophagy,which is induced by AngⅡ.

Objective To study the changes of renal cortex angiotensin Ⅱ (AngⅡ) expression and podocyte autophagy in aging rats with normotension versus hypertension,to further investigate the possible mechanism of renal injury in hypertension during aging.Methods Normotensive Wistar-Kyoto-rats (WKYs) were allocated to three groups by age of 3-month-old group,13-month-old group,22-month-old group.Gender,age and number-matched spontaneously hypertensive rats (SHRs) served as controls.Blood pressure was monitored.Levels of urinary albumin,urinary creatinine,serum creatinine (SCR),blood urea nitrogen (BUN),Ang Ⅱ in serum and in renal cortex were detected.The kidney ultrastructural changes and podocyte autophagosomes were observed under light and transmission electron microscopy.Expressions of nephrin,LC3B Ⅱ,Atg5 and p62 in glomeruli were analyzed by Western blotting.Results Blood pressure was significantly higher in SHRs than in WKYs (P＜0.05).During aging from 3-month-old to 13-month-old and to 22 monthold group,the urinary albumin/creatinine ratio was increased significantly in SHRs with hypertension and in WKYs with normotension [WKY:(0.12±0.01) g/mmol,(0.14±0.04) g/mmol vs.(0.34± 0.05) g/mmol;in SHR:(0.29±0.04) g/mmol,(0.31±0.05) g/mmol vs.(0.72±0.16) g/mmol,P＜0.05],but SCR and BUN levels had no significant difference in SHRs and WKYs during aging (both P＞0.05).And Ang Ⅱ levels in both serum and renal cortex had no significant change in normotension group during aging three time points (both P＞0.05).An age-dependent decrease of Ang Ⅱ level in renal cortex appeared during aging three time points in hypertensive rats [SHR:(0.02 ±0.00) μg/L,(0.02±0.00) μg/L vs.(0.01±0.00) μg/L,P＜0.05],but serum level of Ang Ⅱ had no significant difference during aging three time points (P＞ 0.05) in hypertension group.The structural changes,including glomerulosclerosis,tubular atrophy and interstitial fibrosis were observed during aging three time points both in normotensive and hypertensive rats,but these pathological changes were more serious in hypertensive rats.Autophagosomes relatively accumulated in aging normotensive rats.The protein expressions of LC3B Ⅱ,Atg5 and p62 were increased in normotensive rats during aging (all P＜0.05).While the protein expressions of nephrin,LC3B Ⅱ,Atg5 and p62 were decreased in aging hypertensive rats (all P ＜ 0.05).Conclusions Ang Ⅱ expression level in renal cortex is decreased during aging in hypertensive rats.Podocyte autophagic activity is relatively decreased during aging in normotensive rats,but is relatively increased during aging in hypertensive rats.Ang Ⅱ-induced podocyte autophagy may be involved in the pathogenesis of renal injury in hypertension during aging.

Objective To observe the effects of 1,25-dihydroxyvitamin D3 on the expressions of transforming growth factor-β1(TGF-β1), fibronectin(FN),and vascular endothelial growth factor(VEGF) in rats with diabetic nephropathy(DN), and to elucidate the protective mechanism played by 1,25-dihydroxyvitamin D3. Methods DN models were estabolished by injecting streptozotoein ( STZ ) into male SD rats, which were divided into TGF-β1 overexpression group, TGF-β1 overexpression plus vitamin D3 group, TGF-β1 low-expression group, TGF-β1 low-expression plus vitamin D3 group, TGF-β1 normal-expression group, and TGF-β1 normal-expression plus vitamin D3 group. After 1,25-dihydroxyvitamin D3 treatment for 37 days, renal function and blood biochemical parameters were evaluated. The morphology and fibrosis of kidney tissues were observed. The expressions of TGF-β1, FN, and VEGF in kidney cortex were measured by immunohistochemistry, realtime PCR, and Western blotting. Results The levels of cholesterol, triglyceride, creatinine,plasma glucose, HbA1C , and 24 h urinary protein were lower in vitamin D3treated groups than those in corresponding control groups(P<0. 05). The degree of renal fibrosis was raised with the increased level of TGF-β1. Vitamin D3 treatment decreased the fibrosis in diabetic kidney. There were significant differences in the mRNA and protein expressions of TGF-β1 in three control groups(P<0. 05). With the increased levels of TGF-β1, the expressions of FN and VEGF were increased. The expressions of TGF-β1, FN, and VEGF were lowered by vitamin D3compared with the corresponding control groups(P<0. 05). Conclusion 1,25-dihydroxy-vitamin D3 may protect the renal tissure in diabetic rats via inhibiting the expressions of TGF-β1, FN, and VEGF in the kidney.

To design a new method to simulate the micro-motion of human body surface due to respiration and heartbeat, and to provide detection object and calibration signal for the bio-radar technology. Precision lin-ear module was used to transform rotational movement to linear displacement, with AC servo motor to precisely control the module's rotation. Ultimately, ultralow-frequency micro-motion was produced with its displacement being quantitatively controlled. A system simulating the micro-motion of human body surface was newly built. Compared with the old system, the new one produced micro-motion with better constancy, and realized quantitative control of the motion's dis-placement. The method lays technological foundation for simulating the micro-motion of human body surface due to respiration and heartbeat and may promote the development of bio-radar technology towards intensive and compre-hensive levels.

Abstract: Objective To investigate the clinical outcomes and influencing factors of mild therapeutic hypothermia for influenza-associated encephalopathy/encephalitis (IAE) in children with different center temperatures, and to provide ideas and references for new mild therapeutic hypothermia scheme. Methods　A total of 115 hospitalized children with IAE who were scheduled to receive mild therapeutic hypothermia in Zhongshan Hospital Affiliated to Xiamen University from January 2019 to February 2022 were collected as subjects. They were randomly divided into two groups, namely, the 33 ℃ group (n=60) and the 35 ℃ group (n=55). The clinical features and clinical outcomes of the two groups were analyzed. Univariate and multivariate logistic regression analysis was performed for 6-month to investigate the factors affecting neurological disability. Results　The baseline indicators after treatment, such as Glasgow Coma Scale (GCS) score, cerebrospinal fluid total protein (CSF-TP), CSF lactate dehydrogenase (CSF-LDH), lymphocyte (Lym), creatine kinase-MB (CK-MB), LDH, and neuron-specific enolase (NSE), revealed no significant differences between the two groups before treatment or after treatment (P>0.05). There was no significant difference between the two groups after treatment in the clinical outcomes including GCS score D-value, time of hospitalization, 6-month neurological disability rate and mRS score, CSF-TP D-value, CSF-LDH D-value, Lym D-value, CK-MB D-value, LDH D-value, NSE D-value, improvement rate of EEG and MRI (P>0.05). Univariate and multivariate logistic regression analyses [OR=1.185, 95%CI (1.026~1.369), P=0.021] indicated that the delay of the onset of mild therapeutic hypothermia treatment was an independent risk factor for neurological disability in children with IAE after mild therapeutic hypothermia treatment of 6 months. Conclusion　There was no significant difference in the clinical outcomes between 33 ℃ and 35 ℃ mild therapeutic hypothermia for children with IAE. Therefore, mild therapeutic hypothermia for children with IAE may not require a strict requirement. Timely receipt of mild therapeutic hypothermia is a key measrue to reduce the risk of neurological disability in children with IAE.