Objective To elucidate the relative level of miR-23a RNA in rectal cancer tissues and cell line as well as the effects of miR-23a on the cell proliferation and apoptosis of rectal cancer cells in vitro.Methods Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was applied in assessment of the transcription of miR-23a in rectal cancer tissues and in vitro cells.The RNA fragment of miR-23a inhibitor and inhibitor NC were synthesized and transfected into SW480 cells.Cell proliferation was evaluated with Cell Counting Kit-8 (CCK-8) assay.The apoptotic rate was analyzed by flow cytometry.The expression of ESRP1 was detected by western blot.Wild-type pGL3-ESRP1-3'UTR (wt-pGL3-ESRP1-3'UTR) or mutant pGL3-ESRP1-3'UTR (mut-pGL3-ES-RP1-3'UTR) plasmids and miR-23a inhibitor RNA fragments or inhibitor NC RNA fragments were co-transfected into HEK293 and SW480 cells,then the Promega dual luciferase reporter gene assay kit was used to examine the dual luciferase activity in SW480 cells.Resuits The relative RNA level of miR-23a was significantly promoted in both rectal cancer tissue samples and SW480 cells.After SW480cells were transfected with miR-23a inhibitor,human rectal cancer cell line SW480 with down-regulation of miR-23a showed significant inhibition of cell proliferation compared with negative control (P =0.000).Furthermore,our data demonstrated clearly that the inhibition of miR-23a promoted apoptosis in SW480 cells (P =0.000).Luciferase assay showed that ESRP1 was a direct target gene of miR -23a.Conclusion The expression of miR-23a is clearly associated with the growth and apoptosis of human rectal cells by targeting ESRP1,whilst miR-23a may be used as a potential therapeutic target for the treatment of rectal cancer in the future.

AIM:To investigate the expression of miR-23a and epithelial splicing regulatory protein 1(ESRP1) in rectal cancer tissues and cell lines as well as their effects on rectal cancer cell viability and apoptosis.METHODS:The relative levels of miR-23a in the rectal cancer tissues and cultured cells were assessed by RT-qPCR.The positive expression of ESRP1 in the rectal cancer tissues and non-cancer tissues was detected by immunohistochemical staining.The sequences of miR-23a inhibitor and inhibitor negative control (NC) were synthesized, and transfected into the SW480 cells.The cell viability was measured by CCK-8 assay.The apoptotic rate was analyzed by flow cytometry.The cell invasion was evaluated by Matrigel counting assay.The expression of ESRP1 was determined by Western blot.The wild-type pGL3-ESRP1-3'UTR (wt-pGL3-ESRP1-3'UTR) or mutant pGL3-ESRP1-3'UTR (mut-pGL3-ESRP1-3'UTR) plasmid and miR-23a inhibitor or inhibitor NC were co-transfected into the HEK293 and SW480 cells.The dual luciferase activity was detected according to Promega dual luciferase reporter gene assay kit instructions.The cell viability and apoptosis were evaluated by CCK-8 assay and flow cytometry analysis, respectively, after the SW480 cells were transfected with ESRP1 mimic or mimic NC.The expression of ESRP1, caspase-3, Smac and X-linked inhibitor of apoptosis protein (XIAP) in the SW480 cells was detected by Western blot.RESULTS:The expression of miR-23a was significantly up-re-gulated in the rectal cancer tissues and cell lines, while the positive expression of ESRP1 was significantly decreased in the rectal cancer specimens.The miR-23a expression was also closely related to lymphnode metastasis and TNM stages of rectal cancer patients.ESRP1 was inversely correlated with miR-23a in the rectal cancer tissues.After transfection with miR-23a inhibitor in human rectal cancer SW480 cells, the down-regulation of miR-23a induced significant inhibition of cell viability as compared with the cells transfected with inhibitor NC (P<0.01).Furthermore, the apoptotic rate induced by the miR-23a inhibitor transfection was markedly higher than that of control (P<0.01).Luciferase assay showed that ESRP1 was a direct target gene of miR-23a.The cell viability and apoptosis were inhibited and promoted, respectively, after transfection with ESRP1 mimic in the SW480 cells.Promoted expression of ESRP1 significantly up-regulated the levels of caspase-3 and Smac as well as down-regulated the expression of XIAP in the SW480 cells.CONCLUSION:The expression of miR-23a is significantly associated with the growth and apoptosis of human rectal cancer cells by targeting ESRP1.miR-23a may be a potential therapeutic target for the treatment of rectal cancer in the future.

Objective To preliminarily understand the genotype characteristics of Toxoplasma gondii in blood of HIV?posi?tive persons in Lincang City,Yunnan Province. Method Two segments of SAG2 gene of T. gondii from blood samples of HIV?positive persons in Lincang City were extracted and amplified by using the nested PCR method and the genotype was identified and compared with the standard strain(Type I)of Toxoplasma gondii. Results Thirty?five SAG2 genes(241 bp)and 35 SAG2 genes(221 bp)of T. gondii were amplified from 170 blood samples of the HIV?positive people,and 4 of each case were selected and digested with enzyme,then 2 aim gene fragments of each case were chosen and compared with the standard strain (Type I)of T. gondii. The digestion of SAG2 gene(241 bp)showed the genotype of the blood samples was Type I or Type II, and the digestion of SAG2 gene(221 bp)confirmed that the genotype was Type I. Conclusion It is preliminarily confirmed that the genotype of T. gondii in blood of HIV?positive persons in Lincang City,Yunnan Province is Type I.

Objective To comparatively analyze Toxoplasma gondii separated from HIV-positive people and RH strain GRA6 gene. Methods By using the nested PCR the amplification of Dali HIV-positive blood samples and RH strains of Toxo-plasma GRA6 genome was performed. The GRA6 gene amplification positive product was selected and the electrophoresis imag-ing was performed by being digested with the Mse I endonuclease and the gene sequences were measured and analyzed. Re-sults The GRA6 gene fragment 800 bp was successfully amplified and about 600 bp and 200 bp bands were got by Mse I. The sequencing results showed that T. gondii GRA6 gene positive samples had 2 nucleotide variation compared with T. gondii strain RH namely 447 base pair at C becoming G and 623 base pair at G becoming T. At 146 bp and 690 bp the Mse I restric-tion sites TTAA were found. Conclusion The preliminary judgment shows that the Dali HIV-positive T. gondii genotype is consistent with RH strain belonging to genotype I.

Objective:To evaluate the cerebral blood flow circulation time (CCT) by contrast-enhanced ultrasound, and to explore the change rule of CCT in different degree of intracranial pressure, so as to provide a new method for non-invasive monitoring of intracranial pressure.Methods:Ten patients with hemorrhagic stroke or acute craniocerebral trauma with increased intracranial pressure were selected from Tangdu Hospital, the Air Force Military Medical University from January to December 2019. Contrast-enhanced ultrasound was performed when the invasive intracranial pressure (iICP) increased (>20 mmHg, iICP increased group) and decreased to normal (≤20 mmHg, iICP normal group), CCT was measured and analyzed. The differences of CCTs between different iICP groups were compared and the relationship between CCT and iICP was analyzed.Results:①The CCT on the lesion sides of the same patients in the iICP increased group was significantly longer than in the iICP normal group[(9.34±2.58)s vs (6.48±1.91)s, P=0.002]. ②When iICP was increased in patients with hemorrhagic stroke or acute craniocerebral trauma, the CCTs of the diseased side and the non-pathological side were not statistically significant [(9.34±2.58)s vs (9.01±3.22)s, P=0.809]. ③Pearson correlation analysis and Spearman rank correlation analysis showed that there were no correlations between patient′s breathing, heart rate, carbon dioxide partial pressure, body temperature, GCS score and CCT (all P>0.05). Age, mean arterial pressure and CCT were moderately correlated ( r=0.518, 0.463 and P=0.023, 0.046, respectively). ④Logistic regression analysis showed that CCT was an independent risk factor related to intracranial hypertension( OR=0.7, 95% CI=0.47-0.95, P=0.036). The area under ROC curve (AUC) predicted by logistic regression was 0.750(0.588~0.912). Conclusions:Contrast-enhanced ultrasound noninvasive assessment of CCT can reflect the intracranial pressure in patients with hemorrhagic stroke or acute traumatic brain injury, and CCT has a predictive value for intracranial hypertension. When the patient has limited conditions for invasive intracranial pressure monitoring, or when the invasive monitoring probe is pulled out but still needs to evaluate intracranial pressure, the change of CCT can provide an effective reference for clinical diagnosis and treatment.

To evaluate the accuracy of transcranial color‐code sonography ( TCCS) in non‐invasive assessment of intracranial pressure( ICP ) . TCCS was used to monitor the cerebral hemodynamic parameters of patients with acute severe traumatic brain injury after decompressive craniectomy and make estimation of the non‐invasive intracranial pressure ( ICPtccs) . Methods A total of 91 patients with acute severe traumatic brain injury involved in this retrospective study were divided into the ICP normal group( ≤22 mm Hg ) and the ICP increased group ( ＞22 mm Hg ) . T he correlation and consistency of middle cerebral artery blood flow parameters and ICPtccs with invasive intracranial pressure ( iICP ) were analyzed . According to Glasgow score ( GCS) ,Patients( GCS 3－8) were divided into acute extremely severe traumatic brain injury( GCS 3 －5) and acute severe traumatic brain injury ( GCS 6 －8 ) . A comparison was made of ROC ( ICPtccs) curve and the area under the curve( AUC) between the two groups were cornpared . Results①No statistical differences were found in cerebral hemodynamic parameters between the side with and without decompressive craniectomy in patients with acute severe traumatic brain injury ( all P ＞0 .05 ) . ②M onitored resistive index ( RI) ,pulsatility index ( PI) and ICPtccs between the normal ICP group and the increased ICP group showed statistically significant differences ( all P ＜ 0 .05 ) ,w hile monitored systolic velocity ,diastolic velocity and mean velocity presented no statistically significant difference ( all P ＞0 .05) . T he correlations between RI ,PI with iICP were low ( r＝ 0 .247 ,0 .221 ; all P ＜ 0 .05 ) ,w hile there was a moderate correlation between ICPtccs and iICP( r ＝0 .417 , P ＜0 .001 ) . ③Bland‐Altman plot showed an overestimation of 2 .3 mm Hg ( 95% CI 0 .00－4 .59 mm Hg ) for ICPtccs compared to iICP . ④T he AUC of Glasgow score ( GCS 3－5 and GCS 6－8) in the two groups were 0 .759 ,0 .781 ( all P ＜0 .05) . All the cut‐off points of ICPtccs were 19 mm Hg ,with a sensitivity of 83 .33% ,81 .82% and a specificity of 64 .86% , 75 .68% ,respectively . Pairwise comparison of two AUCs showed no statistical difference ( P ＝ 0 .476) . ICPtccs presented the same ability to estimate ICP in patients with acute severe and extremely severe traumatic brain injury . TCCS could accurately assess the elevation of ICP in 72 .52% patients with acute severe traumatic brain injury . Conclusions TCCS can be used as a non‐invasive screening tool to assess w hether ICP of patients with acute severe traumatic brain injury is elevated and to semi‐quantitatively estimate ICP ,showing useful clinical value .

Objective This study was to develop a"Spleen Deficiency Certificate Scale for County Residents"and test its reliability.It was then developed as an objective tool for Chinese medicine evidence and symptoms for the prevention and control of chronic diseases among county residents.Methods The scale was compiled based on the team's previous foundation.The reliability of the scale was evaluated using internal consistency reliability and split-half reliability,while its validity was evaluated using structural validity,content validity,calibration validity,and discriminant validity.Results The study included 213 adults from Lanxi,of whom 155 were tested for intestinal flora.Seven scale entries were identified:Fatigue,fear of cold,bland mouth,loss of appetite,diarrhea,weak bowel movements,and tooth-marked tongue.In the reliability test,Cronbach's alpha coefficient was 0.828 and McDonald's ω coefficient was 0.825.The"stomach pain"and"bloating"entries did not meet the inclusion requirements and were recommended to be deleted.The Spearman-Brown coefficient was 0.839.The exploratory factor analysis of the two common factors explained 61.6%of the cumulative variance.The calibration validity indicated that the ratio of salivary amylase activity before and after acid stimulation was 0.826±0.253 in the group with spleen deficiency.Significant differences(P<0.05)in the genera Dialister,Shigella,Leuconostoc,Photobacterium,Trabulsiella,and Parvimonas between the spleen deficiency group and the non-spleen deficiency group.Conclusion The Spleen Deficiency Scale for County Residents demonstrates good reliability and validity.

Reducing the level of emotional expression conflict of patients is of great significance to promote the functional recovery of breast cancer patients,improve their prognosis and improve their quality of life.We introduce the research progress of ambivalence over emotional expression in breast cancer patients in terms of the connotation,measurement tools,influencing factors,and intervention methods.There is still a lack of research on measuring the emotional expression conflict of breast cancer patients in China,and it is still necessary to develop a measurement tool with cultural characteristics to obtain the true level of emotional expression conflict of patients.In clinical practice,complementary and alternative medicine,cognitive behavioral intervention,mindfulness training,expressive writing intervention,and other intervention strategies have achieved certain results in improving patients'negative emotions,but personalized intervention strategies still need further exploration.

Objective To observe the effect of Qixian Tongluo Formula on contralateral corticospinal tract(CST)remodeling and motor functional recovery in rats with cerebral infarction,and to explore its potential molecular mechanism from the perspective of regulating factors related to never remodeling.Methods The rat middle cerebral artery occlusion(MCAO)model was established by silk thread ligation.Fifty model rats were randomly divided into model group,citicoline group(0.054 g·kg-1),Qixian Tongluo Formula low-,medium-and high-dose(7.83,15.66,31.32 g·kg-1)groups,and sham operation group,with 10 rats in each group.The intervention administration was started on the 3rd day after operation once a day for 26 consecutive days.On the 3rd,14th and 28th day after operation,the gross motor function was evaluated by Longa score,and the fine motor function was evaluated by beam-walking test(BWT)score.The contralateral motor cortex was injected with the nerve tracer biotin dextran amine(BDA)on the 14 th day after operation to anterogradely trace the CST.On the 28th day after operation,the expression of axonal growth associated protein-43(GAP-43)and BDA positive fibers in the contralateral motor cortex and cervical spinal cord were detected by immunohistochemistry.The co-localization areas of BDA positive fibers and presynaptic marker protein vesicular glutamate transporter 1(VGLUT1)in the cervical spinal cord gray matter were detected by immunofluorescence.The expressions of brain-derived neurotrophic factor(BDNF),glial cell-derived neurotrophic factor(GDNF),nerve growth factor(NGF)and nerve remodeling-associated inhibitory factor[Nogo-A,oligodendrocyte myelin glycoprotein(OMgp)and myelin-associated glycoprotein(MAG)]in the contralateral motor cortex were detected by Western Blot.Pearson correlation analysis was used to analyze the correlation between Longa score or BWT score and BDA/VGLUT1 co-localization area,respectively.Results Compared with the sham operation group,rats in the model group had obvious symptoms of motor function deficits,and the Longa scores were significantly increased(P＜0.01)and the BWT scores were significantly decreased(P＜0.05,P＜0.01)at each time point.The expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was up-regulated(P＜0.05),the number of edge-crossing fibers from the posterior funiculus in cervical cord was increased(P＜0.05),the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was increased(P＜0.05),the expressions of BDNF,GDNF and NGF in the contralateral motor cortex were up-regulated(P＜0.05),while the expressions of Nogo-A,OMgp and MAG were down-regulated(P＜0.05).Compared with the model group,the Longa scores in each administration group on the 14th and 28th day after MCAO operation were significantly decreased(P＜0.01),the BWT scores were significantly increased(P＜0.01),the expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was significantly up-regulated(P＜0.01).The number of edge-crossing fibers from the posterior funiculus in cervical cord was significantly increased(P＜0.01),the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was significantly increased(P＜0.01).The expressions of BDNF,GDNF and NGF in the contralateral motor cortex were significantly up-regulated(P＜0.01,P＜0.05),while the expressions of Nogo-A,OMgp and MAG were significantly down-regulated(P＜0.05),and the most significant effect was observed in the high dose group.The Longa score was negatively correlated with the co-localization area of BDA/VGLUT1(r=-0.89,P＜0.01),and the BWT score was positively correlated with the co-localization area of BDA/VGLUT1(r=0.84,P＜0.01).Conclusion Qixian Tongluo Formula can improve motor function through promoting contralateral CST remodeling in MCAO rats after cerebral infarction,and the molecular mechanism may be related to the regulation of the expression of nerve remodeling-associated factor in the contralateral motor cortex.

Objective:To explore the status quo and influencing factors of death anxiety among middle-aged and young adult patients with chronic heart failure (CHF) .Methods:Totally 176 middle-aged and young adult CHF patients treated at Fuwai Central China Cardiovascular Hospital between January 2021 and February 2023 were selected by convenience sampling and investigated with a general information questionnaire, the Medical Coping Modes Questionnaire (MCMQ), and the Death Anxiety Scale.Results:A total of 176 questionnaires were distributed, with 170 valid responses, yielding an effective response rate of 96.59%. Among the 170 middle-aged and young adult CHF patients, 136 had low death anxiety, while 34 had high death anxiety. Multiple linear regression analysis showed that educational level, per capita monthly family income, duration of illness, cardiac function classification, physical exercise, and coping style (yielding) were influencing factors of death anxiety in middle-aged and young adult CHF patients ( P<0.05) . Conclusions:Death anxiety in middle-aged and young adult CHF patients is primarily associated with educational level, per capita monthly family income, duration of illness, cardiac function classification, physical exercise, and coping style (yielding). Clinical assessments should focus on these factors to provide timely and targeted psychological interventions.