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 the clinical application effect of double eyelid plasty using the levator palpebrae superioris aponeurosis fiber flap.Methods:In March 2021, 3-mm-wide tissue strips were vertically harvested from the mid-upper eyelids from 5 adult Han cadavers with single eyelids, provided by the Anatomy Lab of Nanchang Vocational Health College, and then Masson staining was performed to track the levator aponeurosis fiber. A descriptive case series study was performed. From February 2021 to February 2022, eligible patients who were prospectively collected from the Outpatient Department of the First Affiliated Hospital of Nanchang University underwent double eyelid plasty. The levator palpebrae superioris aponeurosis fiber flap was dissected and repositioned between the orbicularis oculi muscles at the upper and lower edge of incision. The surgical site was closed by a three-layer multi-faceted suture fixation: the muscle of orbicularis oculi at lower edge of incision was sutured to the flap root and tarsal upper edge; the upper and the muscle of orbicularis oculi at the upper and lower edge were sutured to the flap end; the skin was sutured to the muscle of orbicularis oculi and flap end. Postoperatively, incision healing and complications were monitored. Twelve months after surgery, two independent attending plastic surgeons evaluated double eyelid height, curvature, symmetry, and incision scar. The patients were asked to evaluate the effect as satisfied, relatively satisfied, or dissatisfied.Results:Masson staining of 5 lateral upper eyelid specimens revealed that in single-eyelid cases, the levator palpebrae superioris aponeurosis diverged into fibers at the white line, penetrated the orbicularis oculi loosely, and extended to the eyelid margin. Fourty female patients with single or hidden double eyelids were included, aged 18-38 years. After 12 months’ follow-up, all patients had good incision healing without major complications. Twelve months after the operation, the result of doctor evaluation was as followed: 40 cases had proper line height and natural curvature; 38 cases had good line retention, 2 cases had shallower lines; 39 cases had good symmetry, 1 case had asymmetry; 40 cases had no sunken scar. The result of patient satisfaction was as satisfied (35 cases), relatively satisfied (2 cases), dissatisfied (3 cases), with a 92.5% (37/40) satisfaction rate.Conclusion:The levator palpebrae superioris aponeurosis fiber flap method can reconstruct upper eyelid tissue, with natural dynamic double eyelids with few complications and high patient satisfaction.

Objective:To validate the validity of the auricular cartilage fish-shaped framework in comprehensive rhinoplasty for Chinese Han females.Methods:The clinical data of Chinese Han female patients with short nose and saddle nasal deformity undergoing primary rhinoplasty at the Department of Plastic Surgery, the First Affiliated Hospital of Nanchang University, from August 2021 to October 2023, were retrospectively analyzed. Patients underwent comprehensive rhinoplasty, involving the implantation of an auricular cartilage fish-shaped framework at the nasal tip and a dorsal nasal implant on the nasal bridge.Postoperative follow-up monitored for complications. At 6-month postoperatively, a satisfaction survey was conducted among the patients, their family, or friends using a scale where: very satisfied with improvement=4, moderately satisfied with improvement=3, Slightly satisfied with improvement=2, No improvement=1. Additionally, two plastic surgeons evaluated the postoperative outcomes using a scale where: excellent=5, very good=4, good=3, fair=2, poor=1.Preoperative and 6-month postoperative photographs were processed using Image J software to measure the following parameters: nasolabial angle, nasofacial angle, nasofrontal angle, columellar lobular angle, nasal height, and nasal length. Paired t-tests were used for statistical analysis, with P < 0.05 considered statistically significant. Results:A total of 18 patients were included, with a mean age of (26.4 ± 7.2) years (range: 18-45 years). The mean postoperative follow-up duration was (23.0 ± 9.3) months (range: 6 months to 3 years). All patients recovered well, with no severe complications occurring.The mean patient satisfaction score regarding the postoperative outcome was 3.50 ± 0.62. The mean satisfaction score from family or friends was 3.33 ± 0.69. The two evaluating plastic surgeons rated the outcomes as 4.17 ± 0.85 and 3.94 ± 0.93, respectively, both falling between the excellent and very good categories on the assessment scale.Statistically significant differences (all P < 0.01) were found between preoperative and 6-month postoperative measurements for all assessed nasal parameters. Nasolabial angle: 99.68° ± 2.95° vs. 95.37° ± 1.09°; nasofacial angle: 29.47° ± 2.99° vs. 32.82° ± 2.23°; nasofrontal angle: 140.72° ± 5.18° vs. 133.79° ± 4.43°; columellar lobular angle: 44.86° ± 3.43° vs. 39.23° ± 6.44°; nasal height: (57.52 ± 0.60) mm vs. (58.41 ± 0.39) mm; nasal length: (50.08 ± 0.54) mm vs. (50.64 ± 0.31) mm. Conclusion:The use of the auricular cartilage fish-shaped framework combined with a prosthesis in rhinoplasty, requiring only unilateral or bilateral auricular cartilage, effectively improved low nose and short nose deformities in the majority of Chinese Han females. This validates the favorable outcome of this framework in comprehensive rhinoplasty for this population.

Objective:To explore the clinical application effect of double eyelid plasty using the levator palpebrae superioris aponeurosis fiber flap.Methods:In March 2021, 3-mm-wide tissue strips were vertically harvested from the mid-upper eyelids from 5 adult Han cadavers with single eyelids, provided by the Anatomy Lab of Nanchang Vocational Health College, and then Masson staining was performed to track the levator aponeurosis fiber. A descriptive case series study was performed. From February 2021 to February 2022, eligible patients who were prospectively collected from the Outpatient Department of the First Affiliated Hospital of Nanchang University underwent double eyelid plasty. The levator palpebrae superioris aponeurosis fiber flap was dissected and repositioned between the orbicularis oculi muscles at the upper and lower edge of incision. The surgical site was closed by a three-layer multi-faceted suture fixation: the muscle of orbicularis oculi at lower edge of incision was sutured to the flap root and tarsal upper edge; the upper and the muscle of orbicularis oculi at the upper and lower edge were sutured to the flap end; the skin was sutured to the muscle of orbicularis oculi and flap end. Postoperatively, incision healing and complications were monitored. Twelve months after surgery, two independent attending plastic surgeons evaluated double eyelid height, curvature, symmetry, and incision scar. The patients were asked to evaluate the effect as satisfied, relatively satisfied, or dissatisfied.Results:Masson staining of 5 lateral upper eyelid specimens revealed that in single-eyelid cases, the levator palpebrae superioris aponeurosis diverged into fibers at the white line, penetrated the orbicularis oculi loosely, and extended to the eyelid margin. Fourty female patients with single or hidden double eyelids were included, aged 18-38 years. After 12 months’ follow-up, all patients had good incision healing without major complications. Twelve months after the operation, the result of doctor evaluation was as followed: 40 cases had proper line height and natural curvature; 38 cases had good line retention, 2 cases had shallower lines; 39 cases had good symmetry, 1 case had asymmetry; 40 cases had no sunken scar. The result of patient satisfaction was as satisfied (35 cases), relatively satisfied (2 cases), dissatisfied (3 cases), with a 92.5% (37/40) satisfaction rate.Conclusion:The levator palpebrae superioris aponeurosis fiber flap method can reconstruct upper eyelid tissue, with natural dynamic double eyelids with few complications and high patient satisfaction.

Objective:To validate the validity of the auricular cartilage fish-shaped framework in comprehensive rhinoplasty for Chinese Han females.Methods:The clinical data of Chinese Han female patients with short nose and saddle nasal deformity undergoing primary rhinoplasty at the Department of Plastic Surgery, the First Affiliated Hospital of Nanchang University, from August 2021 to October 2023, were retrospectively analyzed. Patients underwent comprehensive rhinoplasty, involving the implantation of an auricular cartilage fish-shaped framework at the nasal tip and a dorsal nasal implant on the nasal bridge.Postoperative follow-up monitored for complications. At 6-month postoperatively, a satisfaction survey was conducted among the patients, their family, or friends using a scale where: very satisfied with improvement=4, moderately satisfied with improvement=3, Slightly satisfied with improvement=2, No improvement=1. Additionally, two plastic surgeons evaluated the postoperative outcomes using a scale where: excellent=5, very good=4, good=3, fair=2, poor=1.Preoperative and 6-month postoperative photographs were processed using Image J software to measure the following parameters: nasolabial angle, nasofacial angle, nasofrontal angle, columellar lobular angle, nasal height, and nasal length. Paired t-tests were used for statistical analysis, with P < 0.05 considered statistically significant. Results:A total of 18 patients were included, with a mean age of (26.4 ± 7.2) years (range: 18-45 years). The mean postoperative follow-up duration was (23.0 ± 9.3) months (range: 6 months to 3 years). All patients recovered well, with no severe complications occurring.The mean patient satisfaction score regarding the postoperative outcome was 3.50 ± 0.62. The mean satisfaction score from family or friends was 3.33 ± 0.69. The two evaluating plastic surgeons rated the outcomes as 4.17 ± 0.85 and 3.94 ± 0.93, respectively, both falling between the excellent and very good categories on the assessment scale.Statistically significant differences (all P < 0.01) were found between preoperative and 6-month postoperative measurements for all assessed nasal parameters. Nasolabial angle: 99.68° ± 2.95° vs. 95.37° ± 1.09°; nasofacial angle: 29.47° ± 2.99° vs. 32.82° ± 2.23°; nasofrontal angle: 140.72° ± 5.18° vs. 133.79° ± 4.43°; columellar lobular angle: 44.86° ± 3.43° vs. 39.23° ± 6.44°; nasal height: (57.52 ± 0.60) mm vs. (58.41 ± 0.39) mm; nasal length: (50.08 ± 0.54) mm vs. (50.64 ± 0.31) mm. Conclusion:The use of the auricular cartilage fish-shaped framework combined with a prosthesis in rhinoplasty, requiring only unilateral or bilateral auricular cartilage, effectively improved low nose and short nose deformities in the majority of Chinese Han females. This validates the favorable outcome of this framework in comprehensive rhinoplasty for this population.

Objective To establish a method for the simultaneous determination of DOX·HCl and LND. Methods HPLC was performed on Agilent 5 HC-C₁₈（2） （4.6 mm × 250 mm, 5 µm） column. The mobile phase was methanol-0.1% TFA aqueous solution, and the gradient elution procedure were: 0 to 3 min, 65% methanol; 3 to 7 min, 65%→90% methanol; 7 to 13 min, 90% methanol; 13 to 15 min, 90%→65% methanol; 15 to 20 min, 65% methanol. The collection time was 20 min, the balance time was 3 min, the UV detection wavelengths were 205 nm and 253 nm. The flow rate was 1.0 ml/min and the column temperature was 35℃. The amount of inlet was 10 µl. Results The method was highly specific, and both DOX·HCl and LND exhibited good linearity in the concentration range of 1-40 µg/ml and 6-240 µg/ml, respectively. The two compounds’ precision, stability, and recovery satisfied the requirements of the method. Conclusion This study established a HPLC method that was suitable for the simultaneous detection of DOX·HCl and LND. This method’s high level of specificity, accuracy, and reliability .

Objective To explore the effect of Lee-Silverman voice treatment (LSVT) on language re-habilitation of dysarthria in Parkinson's disease (PD).Methods A total of 84 patients with PD dysarthria ad-mitted to our hospital from February 2019 to February 2022 were selected as the research objects.According to the random number table method,they were divided into observation group and control group,with 42 cases in each group.The control group received routine speech rehabilitation training,and the observation group re-ceived LSVT for four weeks.The scores of Western Aphasia Battery (WAB) and Aphasia Quotient (AQ) be-fore and after intervention were compared between the two groups,and the differences of electroglottographic parameters such as jitter,F0,shimmer,normalized noise energy (NNE) and harmonic noise ratio (HNR) were compared between the two groups.Results Before intervention,there was no significant difference in WAB scores (listening comprehension,repetition,spontaneous speech,naming) and AQ scores between the two groups (P>0.05).After intervention,the above scores of the two groups were significantly improved,and the observation group was higher than the control group,the difference was statistically significant (P<0.05). Before intervention,the jitter,F0,shimmer,NNE,HNR of vowels "a""i""u" in the two groups were not sta-tistically significant (P>0.05).After intervention,the above-mentioned electroglottic parameters of the two groups were significantly improved.The jitter and NNE of vowels "a""i""u" in the observation group were lower than those in the control group,and the HNR was higher than that in the control group,the differences were statistically significant (P<0.05).Conclusion LSVT can improve the electroglottographic parameters and speech function of patients with PD dysarthria,and the language rehabilitation effect of PD dysarthria is obvious.

Objective:To investigate the application value of double contrast-enhanced ultrasonography (DCEUS) combined with serum pepsinogen (PG) in the diagnosis of early gastric cancer(EGC).Methods:Eighty-two patients suspected of EGC from July 2020 to July 2022 in the People′s Hospital of Guangxi Zhuang Autonomous Region, and preoperative DCEUS examination and PG test were performed, and the patients were divided into benign lesion group(13 cases), early gastric cancer group(57 cases) and progressive gastric cancer group(12 cases) using postoperative pathology as the gold standard. Parameters for comparison included time to peak (TTP), peak intensity (PI), enhanced intensity (EI), serum pepsinogen Ⅰ (PGⅠ), serum pepsinogenⅡ (PGⅡ) and their ratio (PGⅠ/PGⅡ). The sensitivity, specificity, and accuracy of DCEUS and PG alone and in combination for the diagnosis of EGC were analyzed by plotting the ROC curve, and its diagnostic value was compared.Results:In the comparison of DCEUS parameters, PI and EI values were higher in the malignant group than in the benign lesion group and TTP was the opposite, with statistically significant differences (all P<0.05). In the comparison of PG detection, PGⅠ and PGⅠ/PGⅡ were lower in the malignant group than in the benign lesions, and lower in the progressive gastric cancer than in the EGC, while PGⅡ was the opposite, with statistically significant differences (all P<0.05). As shown by the ROC curve results, the sensitivity of DCEUS and PG alone and in combination for the diagnosis of EGC was 80.7%, 73.7% and 87.7%, respectively; the specificity was 76.0%, 72.0% and 80.0%, respectively; and the accuracy was 79.3%, 73.2% and 85.4%, respectively. The area under curve (AUC) of the two modalities alone and combined were 0.784 (95% CI=0.669-0.898), 0.728 (95% CI=0.606-0.850) and 0.839 (95% CI=0.734-0.943), respectively, and the combined diagnosis had a higher diagnostic value than the single diagnostic modality. Conclusions:The combined diagnostic modality of DCEUS and PG can further improve the diagnostic efficacy of EGC and reduce its underdiagnosis rate, which has good application value.

Immunotherapies based on immune checkpoint blockade (ICB) have significantly improved patient outcomes and offered new approaches to cancer therapy over the past decade. To date, immune checkpoint inhibitors (ICIs) of CTLA-4 and PD-1/PD-L1 represent the main class of immunotherapy. Blockade of CTLA-4 and PD-1/PD-L1 has shown remarkable efficacy in several specific types of cancers, however, a large subset of refractory patients presents poor responsiveness to ICB therapy; and the underlying mechanism remains elusive. Recently, numerous studies have revealed that metabolic reprogramming of tumor cells restrains immune responses by remodeling the tumor microenvironment (TME) with various products of metabolism, and combination therapies involving metabolic inhibitors and ICIs provide new approaches to cancer therapy. Nevertheless, a systematic summary is lacking regarding the manner by which different targetable metabolic pathways regulate immune checkpoints to overcome ICI resistance. Here, we demonstrate the generalized mechanism of targeting cancer metabolism at three crucial immune checkpoints (CTLA-4, PD-1, and PD-L1) to influence ICB therapy and propose potential combined immunotherapeutic strategies co-targeting tumor metabolic pathways and immune checkpoints.
Humans ; Antibodies, Monoclonal/pharmacology* ; B7-H1 Antigen/antagonists & inhibitors* ; CTLA-4 Antigen/antagonists & inhibitors* ; Immune Checkpoint Inhibitors/pharmacology* ; Neoplasms/drug therapy* ; Programmed Cell Death 1 Receptor ; Tumor Microenvironment

Inflammatory bowel disease (IBD) is a disorder of immune and microbial dysregulation, induced by environmental factors in a genetically predisposed individual. Studies on IBD-associated genes have demonstrated that Paneth cells (PC) are central to the pathogenesis of IBD, especially in the development of Crohn′s disease (CD). The morphology, quantity and function of PC are easily affected by susceptible genes. In the intestine, PC participate in the formation of stem cell niche and secrete antimicrobial particles, which plays a vital role in regulating intestinal microbiota and maintaining intestinal mucosal homeostasis. Meanwhile, the proliferation and maturation of PC are also dependent on the symbiotic microbiota in the intestine. This review focuses on the interaction of susceptible genes, PC and intestinal microbiota, as well as their influence on the occurrence and development of IBD.