Hepatocellular carcinoma （HCC） is one of the most common malignant tumors worldwide， and most patients are in the advanced stage at the time of diagnosis， leading to a poor prognosis and a low survival rate. Traditional treatment methods often have limited efficacy， while gene therapy， as an emerging therapeutic strategy， has shown great potential with numerous challenges. This article mainly introduces the latest advances in the field of gene therapy for HCC and analyzes the key issues that need to be addressed in the future development of this field. Gene therapy has become a research hotspot in HCC due to its advantages of strong targeting， diverse mechanisms of action， and individualized treatment. Although gene therapy is still in the exploratory stage in the treatment of HCC， some preliminary research findings have been achieved. According to the reports in the literature， the combination of gene therapy with other treatment methods， such as chemotherapy and immunotherapy， can improve treatment outcomes and reduce the side effects of monotherapy. With the continuous advances in technology， gene therapy is expected to bring new hope to HCC patients.

Diabetic cardiomyopathy (DCM) is a medical condition characterized by cardiac remodeling and dysfunction in individuals with diabetes mellitus. Sarcoplasmic reticulum (SR) and mitochondrial Ca²⁺ overload in cardiomyocytes have been recognized as biological hallmarks in DCM; however, the specific factors underlying these abnormalities remain largely unknown. In this study, we aimed to investigate the role of a cardiac-specific long noncoding RNA, D830005E20Rik (Trdn-as), in DCM. Our results revealed the remarkably upregulation of Trdn-as in the hearts of the DCM mice and cardiomyocytes treated with high glucose (HG). Knocking down Trdn-as in cardiac tissues significantly improved cardiac dysfunction and remodeling in the DCM mice. Conversely, Trdn-as overexpression resulted in cardiac damage resembling that observed in the DCM mice. At the cellular level, Trdn-as induced Ca²⁺ overload in the SR and mitochondria, leading to mitochondrial dysfunction. RNA-seq and bioinformatics analyses identified calsequestrin 2 (Casq2), a primary calcium-binding protein in the junctional SR, as a potential target of Trdn-as. Further investigations revealed that Trdn-as facilitated the recruitment of METTL14 to the Casq2 mRNA, thereby enhancing the m⁶A modification of Casq2. This modification increased the stability of Casq2 mRNA and subsequently led to increased protein expression. When Casq2 was knocked down, the promoting effects of Trdn-as on Ca²⁺ overload and mitochondrial damage were mitigated. These findings provide valuable insights into the pathogenesis of DCM and suggest Trdn-as as a potential therapeutic target for this condition.
Animals ; Diabetic Cardiomyopathies/pathology* ; RNA, Long Noncoding/genetics* ; Myocytes, Cardiac/metabolism* ; Mice ; Calsequestrin/genetics* ; Calcium/metabolism* ; Male ; Sarcoplasmic Reticulum/metabolism* ; Methyltransferases/metabolism* ; Mice, Inbred C57BL ; Mitochondria, Heart/metabolism* ; Disease Models, Animal ; Mitochondria/metabolism*

To evaluate the effectiveness and safety of 8-ring assisted traction in endoscopic submucosal dissection (ESD), a total of 13 patients with 15 gastric lesions who underwent 8-ring traction-assisted ESD in Nanjing Drum Tower Hospital from July 2021 to January 2022 were included in the retrospective cohort study. All patients successfully completed the operation. The median procedure time was 56 min, and the dissected area per unit time was 0.20±0.02 cm 2/min. The en bloc resection rate, the complete resection rate and the curative resection rate were 93.3% (14/15), 80.0% (12/15) and 80.0% (12/15), respectively. There were no ESD-related complications, such as delayed bleeding or intraperative and postoperative perforation. No deaths occurred. As a novel internal traction technology, 8-ring traction method is simple to operate, providing a good field of vision during ESD to improve the dissection efficiency and reduce complications, which is worthy of further clinical research and promotion.

Objective:To evaluate the efficacy and safety of endoscopic submucosal dissection (ESD) for circular superficial esophageal cancer.Methods:A retrospective analysis was conducted on 74 consecutive cases of circular superficial esophageal squamous cell carcinoma treated with ESD at Nanjing Drum Tower Hospital from January 2015 to December 2019. The success rate of ESD, curative resection rate, incidence of complications, and additional treatment were mainly observed.Results:One case was transferred to surgery, and the remaining 73 cases successfully completed ESD treatment. The success rate of ESD was 98.6%. Postoperative pathology of ESD revealed that 39 cases achieved curative resection, with a curative resection rate of 53.4% (39/73). Intraoperative muscle layer injury occurred in 15 cases (20.5%), and intraoperative perforation occurred in 1 case (1.4%). Two cases (2.7%) experienced delayed bleeding, and one case (1.4%) experienced delayed perforation. Eleven cases were lost to follow-up, and the remaining 62 cases received follow-up for 36.4±19.0 months. Among the follow-up cases, 12 underwent additional surgery and 5 cases additional chemotherapy and radiotherapy. Among the 57 patients with follow-up data who did not underwent surgery, 49 developed esophageal stenosis after ESD, with an incidence rate of 86.0%.Conclusion:ESD for circular superficial esophageal cancer is generally safe, but it is prone to muscle layer injury during the operation, with a low curative resection rate, a high incidence of postoperative esophageal stenosis, and a high proportion of additional surgical procedures.

Objective:To assess the safety and efficacy of single metal clip traction-assisted endoscopic submucosal dissection (ESD) for the treatment of duodenal lesions.Methods:Data of 45 patients with duodenal lesions who underwent ESD in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School between April 2021 and October 2022 were retrospectively recruited. Among them, 19 patients received single metal clip traction- assisted ESD while 26 patients received traditional ESD. The incidence of complications, dissection speed, en bloc resection rate and R0 resection rate of the two groups were mainly observed. Secondary observation indexes included specimen longer diameter, shorter diameter and area.Results:All 45 procedures were successfully completed, with the procedure time of 44.0 (27.0, 67.0) min for the single metal clip traction- assisted ESD group and 34.0 (24.0, 43.5) min for the traditional ESD group ( Z=-1.678, P=0.093). In the single metal clip traction-assisted ESD group, 2 cases (10.5%) had complications, including 1 intraoperative perforation and 1 postoperative bleeding (approximately 20 mL). There were three cases (11.5%) of complications in the traditional ESD group, including 1 case of postoperative bleeding (approximately 50 mL) and 2 cases of postoperative perforation, with no significant intergroup variation ( P=1.000). The dissection speed of the single metal clip traction-assisted ESD group was 16.0 (11.0, 25.8) mm 2/min, significantly larger than that of the traditional ESD group [5.3 (2.2, 21.1) mm 2/min, Z=-2.287, P=0.022]. The en block resection rate and R0 resection rate of the single metal clip traction-assisted ESD group were both 100.0% (19/19). Additionally, the specimen longer diameter, shorter diameter and area of the single metal clip traction-assisted ESD group were 34.0 (22.0, 45.0) mm, 25.0 (20.0, 34.0) mm, and 745.8 (380.0, 1 342.4) mm 2, respectively, significantly larger than those of the traditional ESD group of 20.0 (12.8, 30.3) mm ( Z=-3.119, P=0.002), 14.0 (8.8, 21.3) mm ( Z=-3.417, P=0.001), 190.4 (84.0, 498.7) mm 2 ( Z=-3.275, P=0.001). Conclusion:Single metal clip traction is safe and effective for duodenal ESD, demonstrating a notable improvement in the dissection speed, especially suitable for large duodenal lesions.

Humans ; Cytomegalovirus ; Cytomegalovirus Infections ; Antigens, Viral ; Immunity ; Autoimmune Diseases ; CD8-Positive T-Lymphocytes

Objective: Hepatitis B surface antigen (HBsAg) loss is seldom achieved with nucleos(t)ide analog (NA) therapy in chronic hepatitis B patients but may be enhanced by switching to finite pegylated-interferon (Peg-IFN) alfa-2a. We assessed HBsAg loss with 48- and 96-week Peg-IFN alfa-2a in chronic hepatitis B patients with partial response to a previous NA. Methods: Hepatitis B e antigen (HBeAg)-positive patients who achieved HBeAg loss and hepatitis B virus DNA < 200 IU/mL with previous adefovir, lamivudine or entecavir treatment were randomized 1:1 to receive Peg-IFN alfa-2a for 48 (n = 153) or 96 weeks (n = 150). The primary endpoint of this study was HBsAg loss at end of treatment. The ClinicalTrials.gov identifier is NCT01464281. Results: At the end of 48 and 96 weeks' treatment, 14.4% (22/153) and 20.7% (31/150) of patients, respectively, who switched from NA to Peg-IFN alfa-2a cleared HBsAg. Rates were similar irrespective of prior NA or baseline HBeAg seroconversion. Among those who cleared HBsAg by the end of 48 and 96 weeks' treatment, 77.8% (14/18) and 71.4% (20/28), respectively, sustained HBsAg loss for a further 48 weeks. Baseline HBsAg < 1 500 IU/mL and week 24 HBsAg < 200 IU/mL were associated with the highest rates of HBsAg loss at the end of both 48- and 96-week treatment (51.4% and 58.7%, respectively). Importantly, extending treatment from 48 to 96 weeks enabled 48.3% (14/29) more patients to achieve HBsAg loss. Conclusion Patients on long-term NA who are unlikely to meet therapeutic goals can achieve high rates of HBsAg loss by switching to Peg-IFN alfa-2a. HBsAg loss rates may be improved for some patients by extending treatment from 48 to 96 weeks, although the differences in our study cohort were not statistically significant. Baseline and on-treatment HBsAg may predict HBsAg loss with Peg-IFN alfa-2a.

Background and purpose:Ca2+plays a very important role in the maintenance of cell biological functions. The storage, release and uptake capacity of Ca2+ is controlled by endoplasmic reticulum (ER). Ca2+ homeo-stasis is essential for cellular energy metabolism and proper protein folding. This study aimed to investigate the effect of cytoplasmic Ca2+ on cisplatin induced ER stress-mediated autophagy in ovarian carcinoma SKOV3 and its underlying mechanism.Methods:The ovarian cancer SKOV3 was used as a study object. The experiment consisted of three parts:① To explore the possible relationship between cisplatin-induced ER stress and autophagy, SKOV3 cells were treated with cisplatin for 0, 6, 12 and 24 h, respectively;② To explore the possible relationship between ER stress induced Ca2+ effux and autophagy, SKOV3 cells were treated with cisplatin for 0, 9 and 12 h, respectively, and TG was used as a positive control;③ To explore the effects of blocking calcium effux on autophagy, SKOV3 cells were divided into control group, cisplatin group, TG group, BAPTA-AM group, cisplatin combined with BAPTA-AM group and TG com-bined with BAPTA-AM group. Western blot was used to detect the protein levels of GRP78 and LC3. Fluo-4 calcium lfuorescent probe was used to examine cytoplasmic Ca2+ levels. Confocal microscopy was used to detect LC3 level by immunolfurescence staining.Results:Compared to control group (0.679±0.011), GRP78 was signiifcantly accumulated at 6, 12 and 24 h after cisplatin treatment and reached the maximum value at 6 h (1.393±0.004,P=0.000). Similarly, compared to control group (0.038±0.000), LC3 puncta were clearly seen after cisplatin treatment and reached the maxi-mum value at 12 h (0.072±0.002,P=0.000). Using confocal microscopy, we found that cisplatin and TG increased LC3 punctate accumulation and cytoplasmic Ca2+ levels in a time-dependent manner. Immunolfuorescent method showed that treatment with cisplatin combined with BAPTA-AM or TG combined with BAPTA-AM increased LC3 punctate accumulation induced by cisplatin or TG. The results of Western blot showed that cisplatin combined with BAPTA-AM (0.071±0.001) or TG combined with BAPTA-AM (0.065±0.001) signiifcantly increased LC3Ⅱ/LC3Ⅰ ratio induced by cisplatin (0.039±0.000,P=0.000) or TG (0.035±0.001,P=0.000).Conclusion:Cisplatin induces intracellular ER stress and autophagy in SKOV3 cells, accompanied by increased cytoplasmic Ca2+ levels. Chelating cytoplasmic Ca2+enhanc-es cisplatin-induced autophagy.

10% and 20% HRP aqueous solution was injected into the anterior wall of the stomach of rabbit, and both the afferent and efferent nerve connections of this region were examined.1. Large number of labeled cells appeared in the dorsal motor nucleus of the vagus nerve (dmnX) on both sides throughout its whole length, except the very rostral and caudal ends of it. The labeled cells were more numerous in the middle part of the nucleus at levels above and below the obex. The distribution of the labeled cells showed certain localization characteristics. Those in the caudal part of the nucleus occupied the medial portion, those in the levels around the obex distributed widely in the nucleus, while in the rostral part they were smaller in number and showed no definite localization.2. In the area between the dorsomedial border of the dmnX and the area pos- trema, the so-called area subpostrema, some disseminated labeled cells, the form of which was similar to that the labeled cells in the dmnX, were also observed.3. Numerous labeled cells appeared in the nodose ganglia on both sides, the number of which was several times more than that in the spinal ganglia, indicating that the afferent connection of the stomach via the vagus nerve predominated over that via the sympathetic nerve. The small-sized labeled cells in the ganglion, however, appeared up to more than 40% of all the labeled cells in it, the functional significance of them, as well as the relation between them and the labeled small cells in the spinal ganglia, is worth further investigation.4. The labeled neurons appeared in the spinal ganglia from Th_3 to L_3, most of them were in Th_3 to Th_(10), showing that the afferents of the stomach via the sympathetic system were diffusely arranged and at the same time concentrated in certain segments.5. Our observations confirmed that the stomach is innervated by elements of bilateral origin. In the vagus system the afferent and efferent neurons appeared almost equally in both dmnX and both nodose ganglia. And in the sympathetic system, the afferent neurons on the left side were 1 to 2 times more than those on the right, i. e., the left side is predominant.