Objective To analyze the effect of LINC00894 on the malignant biological behavior of liver cancer cells by regulating the microRNA(miR)-495-3p/RAS associated binding protein 23(Rab23)axis.Methods Cancer and adjacent tissues of 53 patients with liver cancer who undergoing surgery from January 2023 to November 2023 were collected.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of LINC00894,miR-495-3p and Rab23 in tissues and in vitro cultured cells Huh7,SMMC-7721,HepG2 and L-02.Cells Huh7 were randomly separated into Huh7 group,NC group,si-LINC00894 group,anti-NC group,and anti-miR-495-3p group.CCK-8,flow cytometry and Transwell were used to compare the cell proliferation,apoptosis,migration and invasion of each group.Western blot was used to detect the expression of Rab23 and proliferation-related proteins in cells.The relationship between LINC00894 and miR-495-3p,miR-495-3p and Rab23 was analyzed and verified by dual luciferase reporter gene assay.Results The mRNA and protein expression of LINC00894 and Rab23 were higher in cancer tissues and cells of liver cancer patients,while the mRNA expression of miR-495-3p was lower(P＜0.05).The apoptosis rate of si-LINC00894 group was higher than that of Huh7 group and NC group,and the absorbance at 24 h,48 h,and 72 h,numbers of migrating and invading cells,and expression of Rab23 in cells were lower than those in Huh7 group and NC group(P＜0.05);the absorbance at 24 h,48 h,and 72 h,numbers of migrating and invading cells,and expression of Rab23 in cells in the anti-miR-495-3p group were higher than those in the anti-NC group and si-LINC00894 group,the apoptosis rate of cells was lower than that in the anti NC group and si-LINC00894 group(P＜0.05).The luciferase activity in the WT-LINC00894+miR-495-3p group was lower than that in the WT-LINC00894+miR-NC group(P＜0.05),the luciferase activity in the WT-Rab23+miR-495-3p group was lower than that in the WT-Rab23+miR-NC group(P＜0.05).Conclusion Downregulation of LINC00894 can promote the expression of miR-495-3p,inhibit the expression of Rab23,and thus exert the inhibitory effect on the malignant biological behavior of liver cancer cells.

OBJECTIVE: To explore the molecular cytogenetic characteristics of three fetuses with psu idic(Y)(q11.22) using a combination of multiple methods. METHODS: A total of 11 000 pregnant women who underwent prenatal diagnosis at the Prenatal Diagnosis Center of Taizhou City from January 2019 to October 2024 were selected as the study subjects. Chromosome karyotype analysis (G-banding) and copy number variation analysis based on next-generation sequencing (NGS) were performed on the amniotic fluid/cord blood samples of the 11 000 fetuses. For cases suspected of Y chromosome abnormalities, C-banding and/or fluorescence in situ hybridization (FISH) and AZF microdeletion testing were additionally conducted. This study has been reviewed and approved by the Medical Ethics Committee of Taizhou Hospital, Zhejiang Province (Ethics No. KL20240860). RESULTS: Among the 11,000 prenatal samples undergoing concurrent karyotype and copy number variation analysis, two fetuses with 45,X/46,X,psu idic(Y)(q11.22) mosaicism and one fetus with 46,X,psu idic(Y)(q11.22) were detected. FISH detection indicated that approximately 66.7% of the cells in fetus 2 exhibited a dicentric Y chromosome, and the metaphase karyotype supported the presence of a pseudodicentric chromosome. AZF testing revealed complete deletion of the AZFb+AZFc regions in fetus 2 and fetus 3. CONCLUSION Conventional G-banding karyotype analysis for psu idic(Y)(q11.22) is prone to misdiagnosis or missed diagnosis. The combined application of chromosome karyotype analysis (G+C banding), copy number variation analysis, and FISH detection in clinical practice can accurately diagnose fetuses with psu idic(Y).
Humans ; Female ; Pregnancy ; Prenatal Diagnosis/methods* ; DNA Copy Number Variations/genetics* ; Adult ; Chromosomes, Human, Y/genetics* ; Karyotyping ; In Situ Hybridization, Fluorescence ; Cytogenetic Analysis/methods* ; Fetus ; High-Throughput Nucleotide Sequencing ; Male

OBJECTIVE: To assess the clinical value of digital PCR (dPCR) for the prenatal diagnosis of common fetal aneuploidies. METHODS: A dPCR-based assay was developed for detecting trisomies 21, 18, and 13. A retrospective analysis was carried out on 173 amniotic fluid samples collected by the Prenatal Diagnosis Center of Taizhou Hospital between January 2017 and December 2023. By using chromosomal karyotyping as the gold standard, the diagnostic performance of the multiplex dPCR system was evaluated in a double-blind manner. This study has been approved by the Ethics Committee of Taizhou Hospital (Ethics No. K20250339). RESULTS: Chromosomal karyotyping has identified 59 cases of trisomy 21, 5 cases of trisomy 18, 2 cases of trisomy 13, 6 cases with chromosomal structural abnormalities or mosaicisms, and 101 cases with a normal karyotype. The dPCR results (Z-score cutoff = 4.0, CI = 99.997%) showed full concordance with karyotyping (sensitivity = 100%, specificity = 100%, Kappa = 1). Among the 6 structurally abnormal or mosaicism samples, dPCR has accurately detected 4 cases, but mis-classified 2 cases of trisomy 21 with very low-level mosaicisms (3.3%, 6.9%, respectively) as normal. CONCLUSION The established multiplex dPCR system demonstrated high diagnostic accuracy for common chromosomal aneuploidies, with results available within 24 hours. It can serve as an efficient supplementary tool to conventional chromosomal karyotyping, providing reliable support for time-sensitive clinical decision-making in prenatal diagnosis.
Humans ; Female ; Pregnancy ; Aneuploidy ; Prenatal Diagnosis/methods* ; Karyotyping ; Retrospective Studies ; Polymerase Chain Reaction/methods* ; Chromosome Disorders/genetics* ; Adult ; Trisomy 13 Syndrome/diagnosis* ; Trisomy 18 Syndrome/genetics* ; Down Syndrome/genetics*

Objective To analyze the effect of LINC00894 on the malignant biological behavior of liver cancer cells by regulating the microRNA(miR)-495-3p/RAS associated binding protein 23(Rab23)axis.Methods Cancer and adjacent tissues of 53 patients with liver cancer who undergoing surgery from January 2023 to November 2023 were collected.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of LINC00894,miR-495-3p and Rab23 in tissues and in vitro cultured cells Huh7,SMMC-7721,HepG2 and L-02.Cells Huh7 were randomly separated into Huh7 group,NC group,si-LINC00894 group,anti-NC group,and anti-miR-495-3p group.CCK-8,flow cytometry and Transwell were used to compare the cell proliferation,apoptosis,migration and invasion of each group.Western blot was used to detect the expression of Rab23 and proliferation-related proteins in cells.The relationship between LINC00894 and miR-495-3p,miR-495-3p and Rab23 was analyzed and verified by dual luciferase reporter gene assay.Results The mRNA and protein expression of LINC00894 and Rab23 were higher in cancer tissues and cells of liver cancer patients,while the mRNA expression of miR-495-3p was lower(P＜0.05).The apoptosis rate of si-LINC00894 group was higher than that of Huh7 group and NC group,and the absorbance at 24 h,48 h,and 72 h,numbers of migrating and invading cells,and expression of Rab23 in cells were lower than those in Huh7 group and NC group(P＜0.05);the absorbance at 24 h,48 h,and 72 h,numbers of migrating and invading cells,and expression of Rab23 in cells in the anti-miR-495-3p group were higher than those in the anti-NC group and si-LINC00894 group,the apoptosis rate of cells was lower than that in the anti NC group and si-LINC00894 group(P＜0.05).The luciferase activity in the WT-LINC00894+miR-495-3p group was lower than that in the WT-LINC00894+miR-NC group(P＜0.05),the luciferase activity in the WT-Rab23+miR-495-3p group was lower than that in the WT-Rab23+miR-NC group(P＜0.05).Conclusion Downregulation of LINC00894 can promote the expression of miR-495-3p,inhibit the expression of Rab23,and thus exert the inhibitory effect on the malignant biological behavior of liver cancer cells.

OBJECTIVE To establish a visualized loop-mediated isothermal amplification(LAMP)for rapid detec-tion of Klebsiella pneumoniae and evaluate the sensitivity and specificity of the detection method.METHODS To-tally4 specific LAMP primers were designed by targeting to rcsA gene of the K.pneumoniae.The reaction condi-tions(temperature,time)and the parameters of the reaction system were optimized,the reaction was made visu-alized by using chimeric fluorescent dye SYBR Green Ⅰ.The optimal reactions conditions and reaction systems were determined based on the results of visualization and agarose gel electrophoresis.The optimized conditions and systems were used to test the sensitivity by diluting DNA template on a 10-fold gradient,meanwhile,the specifici-ty of the method was evaluated by detecting K.pneumoniae and other ten species of common bacteria and fungi.RESULTS The optimal reaction temperature was 63 ℃ after the optimization,with the reaction time 35 min;the concentration of buffer solution in the reaction system was determined as 0.8×,with the concentration of magne-sium ion 8 mmol/L,the concentration of dNTPs 1.4 mmol/L,the ratio of internal to external primers 6∶1,the concentration of Bst DNA polymerase 0.32 U/μl,the concentration of betaine 0.75 mmol/L.For the test of sensi-tivity,the method could detect the template with the concentration of 1.5 ng/μl.For the test of specificity,only the detection of K.pneumoniae could display positive visualized result and positive electrophoretic band.CONCLUSIONS The visualized LAMP with high specificity and sensitivity for rapid detection of K.pneumoniae is successfully established,facilitating the observation of the detection result without the use of precise instruments.It is suitable for the detection in grass-root laboratories after successful preliminary application in clinical detection.

OBJECTIVE To establish a visualized loop-mediated isothermal amplification(LAMP)for rapid detec-tion of Klebsiella pneumoniae and evaluate the sensitivity and specificity of the detection method.METHODS To-tally4 specific LAMP primers were designed by targeting to rcsA gene of the K.pneumoniae.The reaction condi-tions(temperature,time)and the parameters of the reaction system were optimized,the reaction was made visu-alized by using chimeric fluorescent dye SYBR Green Ⅰ.The optimal reactions conditions and reaction systems were determined based on the results of visualization and agarose gel electrophoresis.The optimized conditions and systems were used to test the sensitivity by diluting DNA template on a 10-fold gradient,meanwhile,the specifici-ty of the method was evaluated by detecting K.pneumoniae and other ten species of common bacteria and fungi.RESULTS The optimal reaction temperature was 63 ℃ after the optimization,with the reaction time 35 min;the concentration of buffer solution in the reaction system was determined as 0.8×,with the concentration of magne-sium ion 8 mmol/L,the concentration of dNTPs 1.4 mmol/L,the ratio of internal to external primers 6∶1,the concentration of Bst DNA polymerase 0.32 U/μl,the concentration of betaine 0.75 mmol/L.For the test of sensi-tivity,the method could detect the template with the concentration of 1.5 ng/μl.For the test of specificity,only the detection of K.pneumoniae could display positive visualized result and positive electrophoretic band.CONCLUSIONS The visualized LAMP with high specificity and sensitivity for rapid detection of K.pneumoniae is successfully established,facilitating the observation of the detection result without the use of precise instruments.It is suitable for the detection in grass-root laboratories after successful preliminary application in clinical detection.

Objective To explore the mechanism of apoptosis induced by diacetyl hexamethylene diamine(CAHB)in adult acute lymphoblastic leukemia Jurkat cells,and to provide theoretical basis for the clinical application of CAHB.Methods Annexin V+/PI-cell rate of Jurkat cells after CAHB induction was analyzed by flow cytometry.The Annexin V+/PI-cell rate was observed after treatment with caspase-9 inhibitor Z-LEHD-FMK.The expressions of apoptosis-related proteins caspase-8,caspase-9 and caspase-3 were observed by Western blotting.Results After CAHB induction,Jurkat cells were reduced in size,cell membrane crinkling,chromatin thickening,nuclear pyknosis or fragmentation,etc.Typical apoptotic bodies could be seen.CAHB induced Jurkat cell apoptosis by activating caspase-9 and caspase-3 in a dose-effect and time-dependent manner.Caspase-9 inhibitors could inhibit apoptosis induction of CAHB to a certain extent.Conclusion CAHB induced Jurkat cell apoptosis was related to caspase-9 and caspase-3 activation.

In March 2020, the outflow of age limited videos from ICU in Spain inspired us to rethink whether there is age discrimination in the allocation of scarce medical resources. This paper frist reflected on the problem of age discrimination caused by this phenomenon from four moral intuitions: the sacred view of life, the quality of life and values, public health ethics and Chinese culture, and then examined whether it is illegal from the legal level, finally pointed out the negative impact on the society, and put forward that taking age as the standard for the allocation of scarce medical resources is not suitable for China’s national conditions.

BACKGROUND:The objective of this retrospective hospital-based study was to describe the epidemiological features of traumatic spinal cord injury(TSCI)in the intensive care unit(ICU)and assess the incidence and possible risk factors for venous thromboembolism(VTE)following TSCI. METHODS:We retrospectively reviewed the medical records of 370 patients with TSCI who were admitted between January 2018 and March 2023.The following parameters were collected:age,sex,body mass index,occupation,underlying diseases,smoking history,education level,etiology of injury,injury segments,American Spinal Injury Association(ASIA)Impairment Scale score,severity of injury,injury severity score(ISS),VTE risk score(Caprini score),treatment,VTE prophylaxis,ICU length of stay,length of hospital stay,concomitant injuries,and complications.Descriptive statistics were used to summarize the demographic and clinical characteristics of the study participants.Logistic regression analysis was used to determine the risk factors for VTE. RESULTS:The mean age of patients with TSCI was 55.5±13.4 years,with a male-to-female ratio of 6.5:1.The leading cause of TSCI was falls from height(46.5%),followed by traffic accidents(36.5%).The cervical spinal cord was the most affected segment,followed by the thoracolumbar region.Among all the patients,362(97.8%)had concomitant injuries.Complications were observed in 255 patients(68.9%)during hospitalization.The incidence rate of VTE was 25.1%.Logistic regression analysis revealed that age(OR=1.721,95%CI:1.207-2.454,P=0.003),mechanical ventilation(OR=3.427,95%CI:1.873-6.271,P<0.001),and non-use of chemical prophylaxis(OR=2.986,95%CI:1.749-5.099,P<0.001)were risk factors for VTE. CONCLUSION:Falls from height and traffic accidents were the main causes of TSCIs in the ICU,especially for male patients with cervical spinal cord injuries.VTE is a frequent complication in patients with TSCI in the ICU.Age,mechanical ventilation,and non-use of chemical prophylaxis were found to be independent risk factors for VTE following TSCI.