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Objective@#An isochronous substitution model was established to explore the association and substitution effect between college students 24 hour activity behavior and physical health, so as to provide specific activity behavior suggestions for college students to improve their physical health.@*Methods@#A stratified random cluster sampling method was used to conduct physical fitness tests and 24 hour activity behavior surveys among 2 794 college students in 12 colleges and universities in Tianjin.Time spent on sedentary behavior(SB), light intensity physical activity(LPA), moderate to vigorous physical activity(MVPA) and sleep(SLP) time. The isochronous method of components was used to explore the relationship between 24 h activity behavior and physical health.@*Results@#Except for 50 m running, MVPA was negatively correlated with BMI Z ( β =-0.62, P <0.05), but positively correlated with other physical fitness indexes ( β =0.34~274.23, P <0.05). LPA was not associated with lung capacity, sitting forward flexion and 50 m running, and negatively correlated with other physical fitness indexes ( β =-14.30- -0.19, P <0.05). SB was negatively correlated with most physical fitness indexes ( β =-11.57- -0.33, P <0.05), but positively correlated with BMI Z ( β =0.45, P < 0.05 ). In addition to lung capacity, SLP was positively correlated with BMI Z , total physical fitness score,1 minute sit-ups, pull ups, 800/1 000 m running, sitting forward flexion, and 50 m running ( β =0.27-11.21, P <0.05), but negatively correlated with long jump ( β =-0.10, P <0.05）. Isochronous substitution showed that the adverse effects of 30 min/d SB and LPA substitution of MVPA were much greater than the beneficial effects of MVPA substitution for corresponding behaviors (total physical score: SB, -0.58 vs 0.47 points; LPA, -0.50 vs 0.38 points).@*Conclusion@#MVPA and SLP have been found to have a positive effect on physical fitness among college students. Therefore, in the process of improving the physical health of college students, ensuring adequate sleep, improving MVPA and reducing SB as much as possible may be one of the effective methods.
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Objective:To establish a disk (CD) microfluidic chip detection platform for the rapid detection of CALR-1 and CALR-2 mutations in patients with cerebral infarction, and summarize its clinical application value.Methods:Based on microfluidic technology and loop mediated isothermal amplification technology, a CD microfluidic chip detection platform for simultaneous detection of CALR-1 and CALR-2 gene mutations were established, and the sensitivity, specificity, repeatability and accuracy of the platform were verified. A total of 124 patients with cerebral infarction treated in Huashan Hospital, Shanghai Medical College, Fudan University from November 2019 to March 2021 were prospectively selected into the experimental group; and 80 healthy subjects were included in the control group. The CALR-1 and CALR-2 gene mutations in anticoagulant peripheral blood samples were detected by the CD microfluidic chip. Each chip could detect 4 samples at the same time and synchronously detect 3 indexes of each sample. The detection results could be obtained after isothermal amplification for 40 min. At the same time, sequencing method was used to verify the test results, and the consistency of the results of the two detection methods was compared.Results:Using this CD microfluidic chip platform, the synchronous amplification of 3 indexes in the sample could be completed within 40 min without the need of thermal circulation, and the whole detection process of the sample could be completed within 60 min. For samples with a high concentration of target nucleic acid, typical positive signals could be visualized after amplification for 10 min, and the test results would be available within 30 minutes after receiving the samples. The detection sensitivity of CD microfluidic chip method for CALR-1 and CALR-2 mutation load concentration was 1.0% and 0.5% respectively. Nonspecific amplification was not observed for the non-target nucleic acid samples, indicating the high specificity of this method. The coincidence rates of intra and inter batch repeatability were 100% (20/20) respectively. Two samples with CALR gene mutation were found in the cerebral infarction group, both of which were CALR-1 mutations (L367fs*46). There was no CALR-1 or CALR-2 mutation in the control group. The detection results of CD microfluidic chip method were completely consistent with the sequencing verification results (100% [204/204]).Conclusions:The CD microfluidic chip method could be used for the detection of CALR-1 and CALR-2 gene mutations in clinical samples of patients with cerebral infarction. This method has the advantages of high detection sensitivity, good detection specificity, fast detection speed and high detection flux, which is helpful to clarify the etiology of patients with cerebral infarction.
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Objective:To observe the expression of microRNA (miRNA)-7159-5p in gastric cancer tissues, and explore its effect on the proliferation and invasion of gastric cancer cells and its molecular mechanism.Methods:The cancer tissues and adjacent tissues of 29 patients with gastric cancer who underwent surgical resection in Hubei 672 Orthopedic Hospital of Integrated Traditional Chinese and Western Medicine from March 2018 to November 2019 were collected. Fluorescence real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-7159-5p in gastric cancer tissues and adjacent tissues, gastric cancer cell lines and normal gastric epithelial cell lines. Select the cell line with the lowest expression of miR-7159-5p, set the control group and the experimental group, and be transfected with control and miR-7159-5p mimics respectively. qRT-PCR was used to detect the expression of miR-7159-5p in transfected cells. The lymphocyte proliferation test (MTS method) was used to detect the proliferation of transfected cells, and the Transwell chamber method was used to detect the invasion activity of transfected cells. The miRNAMap database and dual luciferase reporter gene experiment were used to predict and verify the target genes of miR-7159-5p. qRT-PCR and Western blot were used to detect the expression of target genes in transfected cells.Results:The expression of miR-7159-5p in gastric cancer tissues was lower than that in adjacent tissues ( P<0.01), the expression of miR-7159-5p in gastric cancer cell lines was lower than that of normal gastric epithelial cells (all P<0.01), and the expression of miR-7159-5p was the lowest in SGC7901 cells ( P<0.01). After transfection, the expression of miR-7159-5p in SGC7901 cells of the experimental group was significantly higher than that in the control group ( P<0.01). After transfection, the proliferation activity and the cell invasion activity of the experimental group was significantly lower than that of the control group (all P<0.01). The target gene of miR-7159-5p was tripartite motif 26 (TRIM26). After transfection, the expression of TRIM26 mRNA in SGC7901 cells of the experimental group was significantly lower than that in the control group ( P<0.01). Western blot showed that after transfection, the expression of TRIM26, c-Myc, cyclin D1, β-catenin protein were lower than those in the control group (all P<0.05). Conclusions:The expression of miR-7159-5p is low in gastric cancer tissues, and miR-7159-5p can inhibit the proliferation and invasion of SGC7901 cells by down-regulating the expression of TRIM26.
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Objective:To investigate the expression of long non-coding RNA (lncRNA) ALMS1-IT1 in colorectal cancer tissues and the molecular mechanism of its effect on the proliferation and migration of colorectal cancer HT-29 cells in vitro.Methods:The cancer tissue specimens and paracancerous tissue (>5 cm from the edge of the tumor) specimens were collected from 40 colorectal cancer patients who were diagnosed by pathological examination after surgical resection in Hubei 672 Orthopedic Hospital of Integrated Traditional Chinese and Western Medicine from July 2018 to November 2020. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of ALMS1-IT1 in colorectal cancer tissues and paracancerous tissues, when the relative expression of ALMS1-IT1 was higher than or equal to its median relative expression, ALMS1-IT1 was highly expressed, and the correlation of ALMS1-IT1 expression with the clinicopathological characteristics of patients was analyzed. HT-29 cells were infected with the empty lentivirus and the lentivirus carrying the ALMS1-IT1 silence sequence, and named control group and si-ALMS1-IT1 group. qRT-PCR was used to detect the expression of ALMS1-IT1 in the two groups of HT-29 cells. CCK-8 method and Transwell experiment were used to detect the proliferation and migration ability of the two groups of HT-29 cells. The starBase v2.0 online database was used to predict ALMS1-IT1 interacting molecules, and qRT-PCR and Western blot were used to detect the expression of these molecules.Results:The relative expression of ALMS1-IT1 in colorectal cancer tissues was higher than that in paracancerous tissues (4.54±0.61 vs. 1.19±0.31, t = 34.89, P < 0.01). The median relative expression of ALMS1-IT1 in cancer tissues of 40 patients was 2.93, and the high expression rate of ALMS1-IT1 was 50.0% (20/40). The high expression rate of ALMS1-IT1 in cancer tissues of TNM stage Ⅲ patients was higher than that in TNM stage Ⅰ-Ⅱ patients, the high expression rate of ALMS1-IT1 in poorly-differentiated patients was higher than that in well- and moderately-differentiated patients, and the high expression rate of ALMS1-IT1 in patients with lymph node metastasis was higher than that in patients without lymph node metastasis (all P < 0.01). The cell proliferation capacity (absorbance value) of HT-29 cells in the si-ALMS1-IT1 group after cultured for 2, 3, 4, and 5 days was lower than that in the control group (all P < 0.05). The number of cell migration at 24 h in HT-29 cells of the si-ALMS1-IT1 group was less than that of the control group (45±7 vs. 112±18, t = 3.45, P < 0.05). Using starBase v2.0 online database to predict that the target gene of ALMS1-IT1 may be miRNA-889-3p (miR-889-3p), and the target gene of miR-889-3p may be ATAD2. Compared with the control group, the relative expression of miR-889-3p in HT-29 cells of the si-ALMS1-IT1 group increased (4.24±0.46 vs. 1.01±0.11, t = 6.81, P < 0.01). Compared with the control group, ATAD2 mRNA ( P < 0.01) and protein expression levels in the si-ALMS1-IT1 group were reduced. Conclusions:ALMS1-IT1 is highly expressed in colorectal cancer tissues, and the ALMS1-IT1 expression is related to the TNM stage, degree of tumor differentiation and lymph node metastasis of patients. Down-regulation of ALMS1-IT1 in vitro may inhibit the proliferation and migration of colorectal cancer HT-29 cells by regulating the miR-889-3p-ATAD2 axis. ALMS1-IT1 may be a therapeutic target for colorectal cancer.
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Objective To establish a novel PCR method that can differentiate the two biotype of Mycoplasma urealytium rapidly based on the disparities of parC gene sequences for clinical routine examination.Methods Design two pairs of specific primers and probes for the targeted gene according to the differences of the parC gene sequences from 14 standard serotypes.The specificity of this amplification was verified by detecting two Mycoplasma urealyticum standard strains including serovar1 and serovar4,50 clinical isolated strains ( 12 are Uu strains and 38 are Up strains) and 7 common bacterias from vagina.Collected 70 swab specimens from patients of Sexually Transmit Department with urogenital inflammation symptom and 71 swab specimens from the gynecology health examination population.All those specimens were detected by culture and our method respectively.The sensitivity of this method was evaluated by comparing with the culture.Differences of the infection rate and distribution of biotypes between different populations were analyzed using statistical software.Results Standard strains of Mycoplasma urealyticum and clinical isolates can be typed into two species by the PCR method without nonspecific amplification.The sensitivity of PCR method is much higher than that of culture ( P＜0.05).The infection rates of Uu,Up and the mixing were 8.57％,61.40％,24.30％ respectively for the patients with vaginitis.However,it was 8.80％,67.60％,8.45％ respectively for the gynecology health examination population.There is a significant difference of the total infection rate between the two population(P＜0.05).It showed no significant difference with the distribution of the two types of simple infection in the two groups ( P＞0.05).But the rate of mixing infection is much higher in the patients with vaginitis ( P＜0.05 ).Conclusion The pathology of Mycoplasma urealyticum may be related to the mixed infection of different biotypes.