ObjectiveThis study aims to explore the mechanism and targets of Shenfu Injection in regulating glycolysis to intervene in myocardial fibrosis in chronic heart failure based on the hypoxia-inducible factor-1α （HIF-1α）/ 6-phosphofructo-2-kinase/fructose-2，6-bisphosphatase 3 （PFKFB3） signaling pathway. MethodsA rat model of chronic heart failure was established by subcutaneous injection of isoproterenol （ISO）. After successful modeling， the rats were randomly divided into the Sham group， Model group， Shenfu injection （SFI， 6 mL·kg^-1） group， and inhibitor （3PO， 35 mg·kg^-1） group， according to a random number table， and they were treated for 15 days. Cardiac function was evaluated by echocardiography， and serum N-terminal pro-brain natriuretic peptide （NT-proBNP） levels were detected by enzyme-linked immunosorbent assay （ELISA）. Fasting body weight and heart weight were measured， and the heart index （HI） was calculated. Pathological changes in myocardial tissue were observed by hematoxylin-eosin （HE） and Masson staining， and the fibrosis rate was calculated. Biochemical assays were used to determine serum levels of glucose （GLU）， lactic acid （LA）， and pyruvic acid （PA）. Western blot was used to analyze the expression of proteins related to the HIF-1α/PFKFB3 signaling pathway （HIF-1α and PFKFB3）， glycolysis-related proteins （HK1， HK2， PKM2， and LDHA）， and fibrosis-related proteins ［transforming growth factor （TGF）-β₁， α-smooth muscle actin （α-SMA）， and Collagen type Ⅰ α1 （ColⅠA1）］. Real-time PCR was used to detect the mRNA expression of HIF-1α and PFKFB3 in myocardial tissue. ResultsCompared with the Sham group， the Model group showed significantly decreased left ventricular ejection fraction （LVEF）， left ventricular shortening fraction （LVFS）， interventricular septal thickness （IVSd）， and interventricular septal strain （IVSs） （P<0.05）， while left ventricular internal dimension at end-diastole （LVDd） and end-systole （LVIDs） were increased （P<0.05）. Serum NT-proBNP levels were significantly increased （P<0.01）， and body weight was decreased. Heart weight was increased， and the HIT index was increased （P<0.05）. Myocardial tissue exhibited inflammatory cell infiltration and collagen fiber deposition， and the fibrosis rate was significantly increased （P<0.05）. Serum GLU was decreased （P<0.05）， while LA and PA levels were increased （P<0.05）. Protein expressions of HIF-1α， PFKFB3， HK1， HK2， PKM2， LDHA， TGF-β₁， α-SMA， and ColⅠA1， as well as the mRNA expression of HIF-1α and PFKFB3 were increased （P<0.05）. Compared with the Model group， both the SFI group and 3PO groups showed significant improvements in LVEF， LVFS， IVSd， and IVSs （P<0.05） and decreases in LVDd， LVIDs， and NT-proBNP levels （P<0.05）. Body weight was significantly increased. Heart weight was significantly decreased， and the HIT index was significantly decreased （P<0.05）. Inflammatory cell infiltration， collagen fiber deposition， and the fibrosis rate were significantly decreased （P<0.05）. Serum GLU levels were significantly increased （P<0.05）， while LA and PA levels were decreased （P<0.05）. Expressions of glycolysis-related proteins， fibrosis-related proteins， and HIF-1α/PFKFB3 pathway-related proteins and mRNAs were significantly suppressed （P<0.05）. ConclusionSFI improves cardiac function in chronic heart failure by downregulating the expression of HIF-1α/PFKFB3 signaling pathway-related proteins， regulating glycolysis， and inhibiting myocardial fibrosis.

Dysregulated RNA splicing is a well-recognized characteristic of colorectal cancer (CRC); however, its intricacies remain obscure, partly due to challenges in profiling full-length transcript variants at the single-cell level. Here, we employ high-depth long-read scRNA-seq to define the full-length transcriptome of colorectal epithelial cells in 12 CRC patients, revealing extensive isoform diversities and splicing alterations. Cancer cells exhibited increased transcript complexity, with widespread 3'-UTR shortening and reduced intron retention. Distinct splicing regulation patterns were observed between intrinsic-consensus molecular subtypes (iCMS), with iCMS3 displaying even higher splicing factor activities and more pronounced 3'-UTR shortening. Furthermore, we revealed substantial shifts in isoform usage that result in alterations of protein sequences from the same gene with distinct carcinogenic effects during tumorigenesis of CRC. Allele-specific expression analysis revealed dominant mutant allele expression in key oncogenes and tumor suppressors. Moreover, mutated PPIG was linked to widespread splicing dysregulation, and functional validation experiments confirmed its critical role in modulating RNA splicing and tumor-associated processes. Our findings highlight the transcriptomic plasticity in CRC and suggest novel candidate targets for splicing-based therapeutic strategies.
Humans ; Colorectal Neoplasms/metabolism* ; RNA Isoforms/metabolism* ; Single-Cell Analysis ; RNA Splicing ; Gene Expression Regulation, Neoplastic ; RNA, Neoplasm/metabolism* ; Transcriptome

Objective:To explore and compare early postoperative clinical outcomes between dynamic and classical superior capsular reconstruction for the treatment of irreparable post-superior massive rotator cuff tears (MIRCTs).Methods:29 patients with MIRCTs treated with autologous fascia dynamic SCR (14) and classical SCR (15) at Department of Sports Medicine of the First Affiliated Hospital of Army Medical University from September 2019 to March 2022 were retrospectively analyzed on preoperative and final follow-up pain visual analogue scale (VAS), Constant-Murley score, American Shoulder and Elbow Surgeons (ASES) score, University of California Los Angeles (UCLA) score, range of motion (ROM), acromiohumeral distance (AHD), Sugaya classification and Goutallier classification.Results:All 29 patients were followed up with an average follow-up time of 36.50±8.18 months for dynamized SCR and 29.33±9.15 months for classical SCR, respectively. There was no significant difference between the two groups in terms of gender, age, course of disease and preoperative AHD ( P>0.05). At the final follow-up, there was no significant difference in the degree of improvement in forward flexion (81.79°±36.14° vs. 69.00°±40.19°, t=0.899, P=0.377), abduction [87.50°(71.25°, 122.50°) vs. 80.00°(45.00°, 95.00°), Z=-1.400, P=0.172] and internal rotation [5.00°(5.00°, 6.00°) vs. 4.00°(1.00°, 6.00°), Z=-0.871, P=0.400]; external rotation improved significantly in the dynamic SCR group compared to classical SCR group [37.50°(30.00°, 41.25°) vs. 25.00°(15.00°, 30.00°), Z=-2.285, P=0.019]. Although both groups showed clinical improvements, no significant difference was found between the dynamic SCR group and the classical SCR group on VAS [4.00(3.75, 5.00) vs. 4.00(3.00, 5.00), Z=-0.029, P=0.949], ASES score (50.99±7.98 vs. 46.47±13.73, t=1.074, P=0.293), Constant-Murley score [62.50(54.00, 69.50) vs. 56.00(47.00, 62.00), Z=-1.956, P=0.112] and UCLA score (20.21±3.53 vs. 18.40±3.87, t=1.315, P=0.199). At the final follow-up, patients in the dynamic SCR group had a higher degree of improvement in AHD (3.66±2.22 mm vs. 2.00±1.75 mm, t=2.247, P=0.033). There was no significant difference in Sugaya grading between the two groups at the final follow-up ( Z=-0.370, P=0.747). As for the degree of improvement in Goutallier's grading, there was an improvement in the dynamic SCR group at the final follow-up versus the preoperative period ( Z=-2.101, P=0.036), while there was no significant difference in the degree of improvement in the classical SCR group at the final follow-up versus the preoperative period ( Z=-0.700, P>0.05). Conclusion:Both dynamic SCR and classical SCR for MIRCTs significantly improved shoulder function. Significant improvements in external rotation, AHD and Goutallier grading were observed in the dynamic SCR group compared to the classical SCR group.

Objective:To explore the relationship between the neural development of preterm infants and gut microbiota.Methods:66 premature infants who were hospitalized in the Neonatology Department of Hunan Children′s Hospital from September 2018 to September 2019 were included in the study. Their fecal samples and clinical data from the first admission were collected. According to the neurodevelopment, the patients were divided into normal neurodevelopment group and neurodysplasia group. The bacterial DNA of fecal samples was extracted by 16S rDNA high-throughput sequencing technology and bioinformatics analysis was conducted to compare the composition and diversity of gut microbiota between the two groups.Results:(1) The Shannon index of gut microbiota in normal neurodevelopmental group and neurodysplastic group was 0.89(0.41, 1.51) and 1.01(0.47, 1.31), respectively. There was no significant difference in diversity index between the two groups ( P>0.05). (2) Bifidobacterium, veronica and negativites in the gut microbiota of the normal neurodevelopmental group were significantly higher (all P<0.05), and streptococcus in the gut microbiota of the dysplastic group were significantly higher ( P<0.05). The gut microbiota of the two groups were mainly enterococcus and escherichia shigella. Conclusions:At the genus level, enterococcus and escherichia are the dominant flora of early gut microbiota in preterm infants. Gut microbiota is related to the neural development of preterm infants. The increased abundance of streptococcus, and the decreased abundance of bifidobacterium, veronicus, and negativites may be risk factors for neurodysplasia of preterm infants. The diversity of gut microbiota in early preterm infants may not be significantly related to neural development.

Objective:To investigate the efficacy and safety of precise mechanical thrombectomy based on ABC 2D scale in acute intracranial large artery occlusion stroke (ALVOs). Methods:A prospective study was performed. Two hundred and two patients with ALVOs accepted early mechanical thrombectomy in Department of Neurology, Maoming Clinical School of Guangdong Medical University from January 2021 to February 2022 were enrolled. They were randomly divided into experimental group ( n=102) and control group ( n=100). Stent retriever partially retracted with intermediate catheter for mechanical thrombectomy (SWIM) was the first choice for patients in control group. ABC 2D scale was used to prejudge the pathogenesis of patients in experimental group: patients with scores of 0-3 were considered as having embolic occlusion and a direct aspiration first pass technique (ADAPT) was the first choice, and SWIM would be chosen if suction catheter could not be in place; patients with scores of 4-7 were considered as having intracranial atherosclerotic stenosis occlusion and SWIM was the first choice. The clinical data, surgical effectiveness, surgical safety, and good prognosis rate 90 d after mechanical thrombectomy (modified Rankin scale scores of 0-2 as good prognosis) of the 2 groups were compared. Results:Experimental group had significantly shorter time from puncture to recanalization (51.0[35.0, 78.5] min vs. 67.0[45.0, 100.0] min), and statistically lower NIHSS scores 24 h after mechanical thrombectomy (10.00[4.75, 16.25] vs. 13.00[8.00, 19.00]), significantly higher good prognosis rate 90 d after mechanical thrombectomy (69.6% vs. 46.0%), statistically lower mortality 90 d after mechanical thrombectomy (3.9% vs. 13.0%) compared with control group ( P<0.05). No significant differences were noted in first-pass effect rate, successful vascular revascularization rate, or incidences of symptomatic intracranial hemorrhage (sICH) and ectopic embolization between the control group and experimental group ( P>0.05). Conclusion:Patients with ALVOs accepted early mechanical thrombectomy can have shorter time from puncture to vascular recanalization and better prognosis after etiologically prejudging by ABC 2D scale for thrombectomy.

Objective To study the invasive histopathological features of papillary thyroid carcinoma(PTC)with tall cell variant(TCV).Methods A retrospective analysis of 19 170 cases of PTC in Ruijin Hospital,Shanghai Jiao Tong University School of Medicine from January 2018 to May 2023 was performed to analyze and the clinicopathological features between TCV-PTC group and classic PTC(cPTC)group.Results Pathological results showed 1 380 cases in TCV-PTC group and 15 578 cases in cPTC group.TCV-PTC had higher proportion and(or)mean value in patients'age,extraglandular invasion,nerve invasion,vascular invasion,maximum diameter of cancer focus,multifocality,lymph node metastasis and BRAF mutation,but the proportion of patients with coexistent Hashimoto's thyroiditis was lower than cPTC,and all results had significant difference(P<0.05).Conclusions Compared with the cPTC,TCV-PTC has stronger local invasive characteristics and lymph node metastasis rate,which provides the basis for the subsequent clinical treatment.

Transglutaminase 2 (TGM2) is a ubiquitous multifunctional protein, which is related to the adhesion of different cells and tumor formation. Previous studies found that TGM2 is involved in the interaction between host cells and viruses, but the effect of TGM2 on the proliferation of influenza virus in cells has not been reported. To explore the effect of TGM2 during H1N1 subtype influenza virus infection, a stable MDCK cell line with TGM2 overexpression and a knockout cell line were constructed. The mRNA and protein expression levels of NP and NS1 as well as the virus titer were measured at 48 hours after pot-infection with H1N1 subtype influenza virus. The results showed that overexpression of TGM2 effectively inhibited the expression of NP and NS1 genes of H1N1 subtype influenza virus, while knockout of TGM2 up-regulated the expression of the NP and NS1 genes, and the expression of the NP at protein level was consistent with that at mRNA level. Virus proliferation curve showed that the titer of H1N1 subtype influenza virus decreased significantly upon TGM2 overexpression. On the contrary, the virus titer in TGM2 knockout cells reached the peak at 48 h, which further proved that TGM2 was involved in the inhibition of H1N1 subtype influenza virus proliferation in MDCK cells. By analyzing the expression of genes downstream of influenza virus response signaling pathway, we found that TGM2 may inhibit the proliferation of H1N1 subtype influenza virus by promoting the activation of JAK-STAT molecular pathway and inhibiting RIG-1 signaling pathway. The above findings are of great significance for revealing the mechanism underlying the interactions between host cells and virus and establishing a genetically engineering cell line for high-yield influenza vaccine production of influenza virus.
Animals ; Cell Proliferation ; Dogs ; Humans ; Influenza A Virus, H1N1 Subtype/genetics* ; Influenza, Human ; Madin Darby Canine Kidney Cells ; Protein Glutamine gamma Glutamyltransferase 2

OBJECTIVE: To assess the practical and health economical values of non-invasive prenatal test (NIPT) in Changsha Municipal Public Welfare Program. METHODS: A retrospective analysis was carried out on 149 165 women undergoing NIPT test from April 9, 2018 to December 31, 2019. For pregnant women with high risks, invasive prenatal diagnosis and follow-up of pregnancy outcome were conducted. The cost-benefit of NIPT for Down syndrome was analyzed. RESULTS: NIPT was carried out for 149 165 pregnant women and succeeded in 148 749 cases (99.72%), for which outcome were available in 148 538 (99.86%). 90% of pregnant women from the region accepted the screening with NIPT. 415 (0.27%) were diagnosed as high risk. Among these, 381 (91.81%) accepted amniocentesis, which led to the diagnosis of 212 cases of trisomy 21 (PPV=85.14%), 41 cases with trisomy 18 (PPV=48.81%) and 10 cases with trisomy 13 (PPV=20.83%). The sensitivity and specificity of NIPT for trisomy 21, trisomy 18 and trisomy 13 were (97.70%, 99.98%), (97.62%, 9.97%) and (100%, 99.97%), respectively. In addition, 213 and 30 cases were diagnosed with sex chromosomal aneuploidies (PPV=46.2%) and other autosomal anomalies (PPV=16.57%), respectively. For Down syndrome screening, the cost and benefit of the project was 120.79 million yuan and 1,056.95 million yuan, respectively. The cost-benefit ratio was 1: 8.75, and safety index was 0.0035. CONCLUSION NIPT is a highly accurate screening test for trisomy 21, which was followed by trisomy 18 and sex chromosomal aneuploidies, while it was less accurate for other autosomal aneuploidies. The application of NIPT screening has a high health economical value.
Aneuploidy ; Cost-Benefit Analysis ; Female ; Humans ; Noninvasive Prenatal Testing ; Pregnancy ; Retrospective Studies ; Trisomy 18 Syndrome/genetics*

Objective:To study the risk factors of metabolic bone disease (MBD) associated fracture in very low birth weight premature infants.Methods:From January 2012 to December 2019, premature infants (gestational age <32 weeks, birth weight <1 500 g) were admitted to our hospital and followed-up regularly for 1.5 years (once every month within first 6 months, then once every 3 months). The infants were assigned into two groups according to X-ray diagnosis: the fracture group and the non-fracture group. The clinical data of the two groups were compared and the risk factors of fracture were analyzed.Results:A total of 62 preterm infants with MBD were included in this study, including 11 in the fracture group and 51 in the non-fracture group. The risk factors of MBD associated fracture included intrauterine growth restriction (IUGR), birth weight <1 000 g, gestational age, respiratory support duration and total parenteral nutrition (TPN) duration ( P<0.05). Logistic regression analysis showed that IUGR ( P<0.05, OR=2.159, 95% CI 1.536~2.759) and TPN duration ( P<0.05, OR=1.143, 95% CI 1.042~1.270) were independent risk factors for fracture. Serum alkaline phosphatase (ALP) in the fracture group was significantly higher than the non-fracture group and 25(OH)VitD was significantly lower than the non-fracture group ( P<0.05). Conclusions:IUGR and TPN duration are risk factors for MBD associated fracture in preterm infants. As biochemical markers of bone metabolism, ALP and 25(OH)VitD levels have clinical value predicting MBD associated fracture.

Objective:To compare the characteristics and differences of intestinal flora in premature infants with late-onset sepsis (LOS) and pneumonia by high-throughput sequencing technology, and to investigate the relationship between intestinal flora and LOS.Methods:Through the case-control method, premature infants with late-onset sepsis who were hospitalized in the neonatal department of Hunan Children′s Hospital from August 2018 to October 2019 were selected as the case group ( n=8). At the same time, premature infants diagnosed with pneumonia were selected as the control group ( n=8). The fecal samples of 16 premature infants were collected for the first time, and the DNA was extracted. The DNA of the target region was amplified by polymerase chain reaction(PCR). High-throughput sequencing was performed using NovaSeq 6000 platform to analyze the composition and diversity of intestinal flora between the two groups. Results:(1) Alpha diversity analysis showed that there was no significant difference in the richness and diversity of intestinal flora between the two groups(all P>0.05). (2) The intestinal flora in premature infants of LOS group and control group were dominated by Firmicutes and Proteobacteria, and facultative anaerobes such as Enterococcus and Escherichia-Shigella were the dominant flora at the genus level. Metastas statistical analysis showed that there was no statistically significant difference in flora composition between the two groups at the phylum level ( P>0.05). (3) Metastas statistical analysis was carried out at the level of class, order, family, genus, and species. The relative abundance of actinomycetes, digestive streptococcaceae and Clostridium in LOS group was higher than that in pneumonia group, and the difference was statistically significant (all P<0.05). (4) The relative abundance of Staphylococcus in the LOS group was significantly greater than that in the control group, but Metastas statistical analysis showed that there was no statistically significant difference in the relative abundance of staphylococcus between the two groups ( P>0.05). (5) Among the 8 cases of LOS, 3 premature infants had positive blood cultures, namely Streptococcus agalactiae, Streptococcus mitis, and Enterococcus faecalis. Enterococcus faecalis belongs to the genus Enterococcus, and Enterococcus belongs to the dominant genus in the LOS group. Conclusions:Different site infections have effects on intestinal microecology of premature infants. There were differences in intestinal flora composition between premature infants with LOS and premature infants with pneumonia.