ObjectiveTo analyze the risk factors of dementia among healthy elderly individuals in the middle of their lives. MethodsA total of 175 participants aged 55 to 75 from two communities in Beijing were included from July, 2021 to April, 2023. Cardiovascular Risk Factors, Aging, and Incidence of Dementia (CAIDE) related risk factors and other demographic data were collected. According to the CAIDE assessment, participants with scores ≥ 9 were as high-risk group, and those with scores < 9 were as low-risk group. They were evaluated with Stroop Color Word Test (SCWT), two elements 1-back task paradigm and the revised Trail Making Test (TMT); measured the grip strength, 30 s forearm flexion tests and five sit-to-stand tests; the average step speed and step length of a 10-meter walk were recorded. ResultsThe average total score of CAIDE was 9.86 in the high-risk group, and was 4.95 in the low-risk group. There was no difference in age between two groups (P = 0.188). There were differences in the proportion of participants of male, less than seven years' education, systolic blood pressure > 140 mmHg, cholesterol > 6.5 mmol/L, body mass index > 30 kg/m², and lack of physical activity between two groups (χ² > 3.116, P < 0.05). The grip strength (t = -4.174), walking speed (t = -2.414), SCWT accuracy (Z = -2.684) were all worse in the high-risk group than in the low-risk group (P < 0.05). Logistic regression analysis showed that walking speed (OR = 25.483), grip strength (OR = 1.133) and SCWT accuracy (OR = 37.430) were independent influencing factors of dementia (P < 0.05). ConclusionWeaker grip strength, slower gait speed and worse inhibitory control might be independent influencing factors of dementia.

Objective:To explore the application value and treatment effects of multiple free anterolateral perforator flaps of calf for reconstruction of multiple digital wounds in single hand.Methods:From August 2020 to March 2022, 12 patients with soft tissue defects in 35 digits were treated in Department of Hand Surgery, Suzhou Ruihua Orthopaedic Hospital. Ten patients were male and 2 were female, aged 25 to 58 years old. Of the patients, 1 had soft tissue defects in 5 digits, 3 in 4 digits, 2 in 3 digits and 6 in 2 digits. The size of defects was from 1.2 cm ×1.2 cm to 7.0 cm×3.5 cm after debridement. The vascular perforators discovered from intraoperative explorations were found originating from the superficial peroneal artery in 24 flaps, from the peroneal artery in 7 flaps and from the anterior tibial artery in 4 flaps. During surgery, the perforator artery and accompanying veins of the flaps were anastomosed with the proper digital artery and palmar or dorsal subcutaneous veins in the recipient site, respectively. The size of the flaps was from 1.5 cm×1.5 cm to 7.5 cm×4.0 cm. No nerve was affected in the surgery. The wound at donor sites in the calf was sutured directly. Regular postoperative follow-ups were conducted at outpatient clinics. The comprehensive evaluation scale of flap was used to assess the conditions of the donor and recipient sites.Results:In this study, all 35 soft tissue defects of digits in 12 patients were reconstructed by the anterolateral perforator flaps of calf. All the 35 flaps survived after surgery, with a 100% of survival rate. The patients were instructed to carry out early functional training after surgery. Follow-up lasted 6 to 24 months, with an average of 11 months. Twenty-five flaps were found in slightly swollen, and further flap thinning surgery were carried out 3 months after the primary surgery, while the rest of the flaps had good appearance and texture. At 6 months after surgery, all flaps recovered a partial deep and shallow sensory and sense of touch. All wound at donor sites in calf had one-stage healing without dysfunction. The comprehensive evaluation scale was excellent in 28 flaps and good in 7 flaps. The excellent and good rate was 100%.Conclusion:It is an effective method to use multiple free anterolateral perforator flaps of calf to reconstruct multiple digit defects in single hand. The flaps can be conveniently harvested and the multiple digital defects in single hand can be reconstructed in primary surgery with small damages to the donor sites and together with satisfactory clinical outcomes.

Objective To analyze the infection characteristics and epidemic trend of 9 respiratory pathogens in Shenzhen popula-tion .Methods The 5918 patients with acute respiratory tract infection were collected ,indirect immunofluorescence was used to de-tect the IgM antibody of 9 respiratory pathogens ,including legionella pneumophila type 1 (LP1) ,mycoplasma pneumoniae (MP) , rickettsia Q (COX) ,rickettsia and chlamydia pneumoniae (CPn) ,adenovirus (ADV) ,respiratory syncytial virus (RSV) ,influenza A virus (INFA) ,influenza B virus (INFB) ,parainfluenza virus (PIVs) ,the results were analyzed statistically .Results A total of 1376 samples were detected at least one pathogen in 5918 serum samples ,the total positive rate was 23 .25% .The positive rate of MP was the highest (15 .19% ) ,followed by the INFB (8 .11% ) .The positive rates of other pathogens were relatively low .The mixed infection positive rate was 4 .29% .The positive rates of MP ,INFB and PIVs in women were significantly higher than those in men(P<0 .001) .The positive rates of MP ,INFB and PIVs were different in different seasons ,the differences were statistically sig-nificant(P<0 .001) .The positive rates of MP ,ADV and INFB in 0 to 14 years old group were significantly higher than those in >14 to 60 years old group and >60 years old group(P<0 .05) .The positive rates of MP ,CPn ,ADV ,RSV ,INFB and PIVs were closely related with age in infant and children (0 to 14 years old)(P<0 .05) .Conclusion The main respiratory pathogens of SARS in Shenzhen city were MP and INFB ,the 9 pathogens had their own infection characteristics and epidemic trend .

Objective To evaluate the effect of microRNA-92a (miR-92a) on regulating cell proliferation by targeting Krüppel-like factor 4 (KLF4) in colon cancer.Methods The miR-92a expressions in 21 colon cancer tissues and matched normal tumor-adjacent tissues and 4 colon cancer cells (HT29,SW480,SW620,HCT116) were detected using quantitative real-time polymerase chain reaction (qRT-PCR).Models of over-expression and suppression of miR-92a were established by transient transfection of miR-92a-3p mimic to HCT116 and transient transfection of miR-92a-3p inhibitor to SW620,respectively.Cell proliferation activity was detected by the CCK-8 colorimetry method,cell cycles were detected by flow cytometry,KLF4 protein expression was detected by Western blotting,and cell luciferase activity was detected by the dual luciferase reporter gene experiment.Results The expression level of miR-92a in colon cancer tissues was (0.648 ± 0.489) fmol/μg total RNA,significantly higher than that in matched normal tumor-adjacent tissues [(0.064 ± 0.062) fmol/μg total RNA],with statistically significant difference (t =-5.420,P ＜ 0.001).In 4 colon cancer cell lines,the miR-92a expression level in HCT116 cells was the lowest,and highest in SW620 cell.When the expression of miR-92a was up-regulated,the cell proliferation activity of 72 h in HCT116 cells was higher than that in the negative control group (0.919 ±0.014 vs.0.765 ± 0.025),with statistically significant difference (t =-9.309,P =0.001),the proportion of S phase cells was also significantly increased [(41.670 ±0.461)％ vs.(38.703 ±0.554)％,t =-7.127,P =0.002),and KLF4 protein expression was decreased (0.460 ± 0.048 vs.0.758 ± 0.109,t =22.865,P =0.028).When the expression of miR-92a was down-regulated,the cell proliferation activity of 72 h in SW620 cells was lower than that in the negative control group (0.608 ± 0.011 vs.0.713 ± 0.005),with statistically significant difference (t =15.920,P ＜ 0.001),while the proportion of S phase cells was decreased [(31.935 ± 0.365) ％ vs.(34.955 ± 0.465) ％,t =8.849,P =0.001],and KLF4 protein expression was increased (0.694 ± 0.121 vs.0.479 ± 0.044,t =-5.246,P =0.034).KLF4 3'UTR wild-type dual luciferase report plasmids were co-transfected with miR-92a-3p mimic to HCT116 cell,and dual luciferase assay showed that miR-92a slightly repressed firefly luciferase actively,but the difference was not statistically significant (t =0.878,P =0.429).There was a negative correlation between the expression of miR-92a and the expression of KLF4 protein in colon cancer tissues,but with no statistical significance (r =-0.163,P =0.699).Conclusion miR-92a is highly expressed in colon cancer tissues.It can promote colon cancer cells proliferation via enhancement of the cell cycle transition of G0-G1 phase to S phase.Up-expression of miR-92a may play a role in down-regulating the expression of KLF4 protein in colon cancer cells.However,KLF4 is not a direct target gene of miR-92a.

Objective To explore the screening of peroxisome pathway reactive oxygen species (ROS) oxidative stress gene and its correlation with the antitumor sensitivity of artesunate against pancreatic cancer. Methods Based on microarray mRNA expressions of 55 tumor cell lines in the National Cancer Institute common database,peroxisome pathway-related key genes which were significant correlation with half-inhibitory concentration (IC50 )values of artesunate antitumor activity against human pancreatic cancer were selected by Kendall test.The candidate genes associated with artesunate sensitivity were identified and their mRNA expressions in pancreatic cancer cells were tested using fluorescent quantitative PCR.The contents of peroxi-dase in pancreatic cancer cells were detected through the DAB staining.Results Thirteen key genes mRNA expressions in peroxidase pathways were significantly correlated with IC50 values for artesunate antitumor activi-ty.Compared with normal liver cells HL-7702 (1.00),CRAT (2.89 ±0.06),PEX11B (1.90 ±0.07)and PEX16 (1.35 ±0.07)mRNA expression levels were significantly increased in pancreatic cancer Panc-1 cells which sensitive to artesunate (t =33.00,P <0.01;t =17.85,P <0.01;t =4.54,P <0.05).While CAT
(1.43 ±0.03),SOD1 (2.07 ±0.04)and SOD2 (1.15 ±0.01)mRNA expression levels were also signifi-cantly increased in Panc-1 cells which sensitive to artesunate (t =11.71,P <0.01;t =35.85,P <0.01;t =13.22,P <0.01).However,PEX12 (0.51 ±0.02),CAT (0.47 ±0.02),PRDX1 (0.43 ±0.01),and SOD1 (0.44 ±0.01)mRNA expression levels in pancreatic cancer BXPC-3 cells which resistant to artesunate were significantly lower than that of HL-7702 cells (t =37.53,P <0.01;t =16.52,P <0.01;t =84.20, P <0.01;t =48.24,P <0.01).DAB staining showed that the positive expression rate of peroxisomal content was apparently higher in Panc-1 cells (61.5%)than that of HL-7702 cells (43.8%),with a significant difference (χ2 =16.11,P <0.01).Conclusion Peroxisome and its related ROS antioxidant enzymes CAT, PRDX1,SOD gene expression may be the important factors that affect artesunate antitumor activity against human pancreatic cancer.

Objective:To investigate the expression of miR-92a in colorectal cancer (CRC) and its effect on the regulation mechanisms of tumor angiogenesis. Methods:The miRNA-92a expression in 25 CRC tissues and HT29, SW620, SW480, and HCT116 CRC cells was detected using quantitative real-time polymerase chain reaction. The CD31 positive expression of blood microvessels in CRC tissues was measured by immunohistochemistry. Pearson correlation analysis was used to analyze the relationship between miR-92a and tu-mor angiogenesis. The miR-92a mimic or inhibitor was transfected into HCT116 and SW620 cells in vitro to upregulate or downregu-late the miR-92a expression. The effect of miR-92a overexpression or suppression on the formation of human umbilical vein endotheli-al cell (HUVEC) tubules was detected by tube formation assay. Changes in phosphatase and tensin homolog (PTEN) protein expression were measured by Western blot. Results:The expression level of miR-92a in CRC tissues was significantly higher than that in matched adjacent tissues (P<0.01). The expression levels of miR-92a in HT29, SW480, SW620, and HCT116 colon cancer cell lines were signifi-cantly higher than that of the normal colorectal epithelium control (P<0.05). The number of CD31 positive expression of microvessel density (MVD) in CRC tissues was significantly higher than that in adjacent tissues (P<0.01), and the miR-92a expression level was posi-tively correlated with the MVD in CRC tissues (r=0.580, P=0.01). The cell culture supernatant of HCT116 with miR-92a overexpression could significantly promote the formation of HUVEC tubules (P<0.05). The upregulation of miR-92a expression could significantly inhib-it the expression of PTEN protein in CRC cells (P<0.01). Conclusion:The miR-92a was highly expressed in CRC cells and tissues, which was closely related to the formation of tumor angiogenesis in CRC. The miR-92a could promote tumor angiogenesis in CRC by inhibit-ing PTEN expression.

Objective To evaluate the value of homocysteine(Hcy) detection by different assays in the diagnosis of hypertension complicating cerebral infarction disease .Methods Serum concentration of Hcy in the patients with single hypertension ,hyperten‐sion complicating cerebral infarction and healthy controls were detected by the fluorescence quantification assay and the enzymatic cycling assay respectively ,the application value of the two assays in the diagnosis of hypertension complicating cerebral infarction disease was evaluated by the receiver operating characteristic (ROC) curve .Results Both the two assays showed that the serum Hcy level and incidence of high Hcy in the single hypertension group and the hypertension complicating cerebral infarction group were obviously higher than those in the healthy control group (P<0 .05) ,but there were no statistically significant differences be‐tween the single hypertension group and the hypertension complicating cerebral infarction group (P>0 .05) .The ROC curve analy‐sis showed that the sensitivity and the specificity in the enzymatic cycling assay were higher ,which were 67 .5% and 85 .3% respec‐tively ,but the fluorescence quantification assay had lower sensitivity (40 .0% ) and higher specificity (97 .1% ) .Compared to the sin‐gle index test ,the sensitivity of Hcy and triglyceride combination detection had higher sensitivity (72 .5% ) and higher specificity (94 .1% ) ,which were higher than the diagnostic performance of the single index .Conclusion The increase of serum Hcy level is closely related to hypertension ,but is not a direct causal relationship with hypertension complicating cerebral infarction .The enzy‐matic cycling assay is better than the fluorescence quantitative assay in the diagnostic performance of hypertension complicating cer‐ebral infarction .The combination detection of Hcy and triglyceride conduces to improve the diagnostic sensitivity and specificity .

Objective To investigate the effect of shRNA-mediated down-regulation of the receptor for activated C kinase 1 (RACK1) gene on the chemotherapeutic sensitivities in human lung adenocarcinoma cell line A549.Methods The shRNA recombinant plasmid targeting to human RACK1 gene was designed and transferred into A549 cells by lipofectin technique.The protein level of RACK1 was measured by Western blot to confirm the function of shRNA plasmid.Drug sensitivities of A549 cells to cisplatin,gemcitabine,pemetrexed and paclitaxel were analyzed by MTT assay.The protein expression of LRP and MRP were detected by Western blot.Results After 24 hours transfection,the relative expression quantity of RACK1 protein in RACK1-shRNA group was 0.267± 0.470,which was significantly lower than that in vector-shRNA group (0.821±0.109) and control group (0.842±0.060) (F =54.438,P ＜ 0.05).The results of MTT showed that the growth of A549 cells in the RACK1-shRNA group was markedly inhibited.The sensitivities of A549 cells to cisplatin and paclitaxel were significantly enhanced compared with that in the vector-shRNA group and control group (P ＜ 0.05).The relative expression quantity of LRP and MRP protein in RACK1-shRNA group were 0.163±0.056 and 0.246±0.050,which were lower than that in vector-shRNA group and control group (F LRP =19.430,F MRP =61.548,both P ＜ 0.05).Conclusion Targeted gene silencing of RACK1 improves the sensitivity of A549 cells to the ascisplatin and paclitaxel medicines,which might be achieved through down-regulation of the expression of LRP and MRP.

Objective To explore the expression characteristic of serum mir-16,mir-21,mir-29a, mir-92a and mir-143 in colorectal cancer , evaluate the clinical significance of candidate miRNAs in CRC diagnosis.Methods Case-control study was used.The expression levels of serum mir-16,mir-21,mir-29a, mir-92a and mir-143 from 50 CRC patients and 27 normal controls were detected by real-time PCR, the levels of miRNAs expression in serum of 8 CRC patients 1 day before and 7 days after radical surgery were analyzed.The sensitivity and specificity of serum miRNAs expression for the diagnosis of colorectal cancer were evaluated using the receiver operating characteristic ( ROC) curve.Results The expression level of mir-16, miR-21, mir-92a in serum of patients with CRC were (75.55 ±37.73), (35.96 ±23.81), (24.79 ±8.97) fmol/ml, significantly higher than that of the healthy control group (32.73 ±18.94), (24.36 ±13.27), (16.36 ±5.58) fmol/ml (tmir-16 =2.77, tmir-21 =2.34, tmir-92a =3.85,P <0.05).However, the expression levels of mir-29a and mir-143 showed no significant difference between two groups (tmir-29a=-0.17, tmir-143 =1.59,P>0.05).In addition, serum miR-16 and miR-92a levels of CRC patients 7 days after tumor resection were (36.02 ±19.95), (14.82 ±7.78) fmol/ml, significantly lower than that before operation (62.18 ±34.17), (24.06 ±12.99) fmol/ml (tmir-16 =3.59, tmir-92a =2.60,P<0.05).Area under the ROC curve ( AUC) of combined detection of mir-16, miR-21and mir-92a was 0.877, the sensitivity and specificity were 88% and 85%, respectively, which was higher than any of 3 miRNAs alone and the conventional tumor marker CEA.Conclusion Combination of mir-16, mir-21 and mir-92a in the diagnosis of CRC shows a better sensitivity and specificity , which would be expected as new tumor biomarkers for noninvasive diagnosis and monitoring progression of colorectal cancer.

ObjectiveTo investigate the biological effects and its mechanisms of ascorbic acid on pancreatic cancer PANC1 cells. Methods PANC1 cells were treated by ascorbic acid of different concentrations (0 ～40 mmol/L) for 24,48,72 hours.The proliferation of PANC1 cells was analyzed by MTT method; cell cycle and apoptosis were assessed by flow cytometry (FCM); inverted microscopy and transmission electron microscopy were used to observe cell morphology. The membrane potential of mitochondria were mearured by with JC-1 staining and FCM.Meanwhile,the changes of cell morphology and mitochondrial membrane potential induced by ascorbic acid after pretreatment with hydrogen peroxidescavenging enzyme (catalase) and red blood cells were also detected. Results Ascorbic acid in pharmacologic concentrations selectively inhibited the proliferation of PANC1 cells in a dose and time dependent manner.PANC1 cells were arrested in G2/M phase after treatment with 5 mmol/L ascorbic acid [ (32.55 ± 7.14)％ vs (22.00 ±1.27)％,t =5.808,P＜0.05],but there was no changes on apoptosis rate [ (1.98 ± 1.80)％ vs (1.09 ±0.16)％ ].Inverted microscope and transmission electron microscopy showed that oncosislike cell death of PANC1 cells was induced after treatment with ≥5 mmol/L ascorbic acid.Mitochondrial membrane potential of PANC1 cells was significantly lower than that of the control group in a dose dependent manner.The descent of mitochondrial membrane potential was significantly inhibited by pretreatment with catalase and red blood cells,and the degree of cell oncosis was attenuated.ConclusionsAscorbic acid significantly inhibited the proliferation of pancreatic cancer PANC1 cells in vitro.Ascorbic acid induced PANC1 cell oncosis,but not apoptosis.The possible mechanisms of inducing oncosis may be related to the descent of mitochondrial membrane potential.