Recent clinical studies have shown that mutation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene in cancer cells may be associated with immunosuppressive tumor microenvironment (TME) and poor response to immune checkpoint blockade (ICB) therapy. Therefore, efficiently restoring PTEN gene expression in cancer cells is critical to improving the responding rate to ICB therapy. Here, we screened an adeno-associated virus (AAV) capsid for efficient PTEN gene delivery into B16F10 tumor cells. We demonstrated that intratumorally injected AAV6-PTEN successfully restored the tumor cell PTEN gene expression and effectively inhibited tumor progression by inducing tumor cell immunogenic cell death (ICD) and increasing immune cell infiltration. Moreover, we developed an anti-PD-1 loaded phospholipid-based phase separation gel (PPSG), which formed an in situ depot and sustainably release anti-PD-1 drugs within 42 days in vivo. In order to effectively inhibit the recurrence of melanoma, we further applied a triple therapy based on AAV6-PTEN, PPSG^@anti-PD-1 and CpG, and showed that this triple therapy strategy enhanced the synergistic antitumor immune effect and also induced robust immune memory, which completely rejected tumor recurrence. We anticipate that this triple therapy could be used as a new tumor combination therapy with stronger immune activation capacity and tumor inhibition efficacy.

AIM:To investigate the effect of circular RNA MYO9A-006(circMYO9A_006)on hypertrophic phenotype of cardiomyocytes and the underlying mechanism.METHODS:The effect of adenovirus-mediated overexpres-sion of circMYO9A_006 on the expression of hypertrophy-related proteins,including β-myosin heavy chain(β-MHC),skeletal muscle actin alpha 1(ACTA1)and atrial natriuretic peptide(ANP),was evaluated in neonatal mouse ventricular cardiomyocytes(NMVCs).Moreover,a neonatal rat ventricular cardiomyocyte(NRVC)model of phenylephrine(PE)-in-duced hypertrophy was established.The effect of circMYO9A_006 overexpression on NRVC size was ascertained using Phalloidin-iFluor 647 staining method.Dual-luciferase reporter assay was employed to measure the activity of potential in-ternal ribosome entry sites(IRES)in circMYO9A_006.The translation and intracellular location of the circMYO9A_006-translated protein,MYO9A-208aa,were verified using Western blot.To investigate the role of MYO9A-208aa in the ef-fect of circMYO9A_006 on the cardiomyocyte hypertrophic phenotype,we prepared and used the following adenoviruses:the recombinant circMYO9A_006-ORF adenovirus to express MYO9A-208aa,the recombinant circMYO9A_006-ATG-mut adenovirus that does not express MYO9A-208aa,the recombinant circMYO9A_006 adenovirus,and the adenovirus vector control.These were then employed to infect NRVCs.RESULTS:Successful adenovirus-mediated overexpression of circMYO9A_006 was observed in NMVCs.The increased expression of circMYO9A_006 notably reduced the expres-sion of hypertrophy-related proteins in NMVCs(P<0.01).Concurrently,overexpression of circMYO9A_006 substantially reduced the expression of hypertrophy-associated proteins and diminished the size of PE-induced NRVCs(P<0.05).Dual-luciferase reporter assay identified the activity of 2 IRES in circMYO9A_006.Western blot results indicated that circ-MYO9A_006 could produce the MYO9A-208aa protein with an anticipated molecular weight of 28 kD in NRVCs,primari-ly found in the cytoplasm.Elevated expression of both circMYO9A_006 and MYO9A-208aa consistently reduced the ex-pression of hypertrophy-associated proteins(P<0.01),and counteracted the enlarged size of PE-induced NRVCs(P<0.05).However,increased expression of circMYO9A_006-ATG-mut did not counteract the PE-induced hypertrophic phe-notype of NRVCs.CONCLUSION:circMYO9A_006 attenuates the hypertrophic phenotype of cardiomyocytes by synthe-sizing the MYO9A-208aa protein.

Objective:This study aims to evaluate the quality and explore the preliminary clinical applications of a domestically developed hepatitis D virus nucleic acid quantification reagent (abbreviated as"domestic HDV RNA reagent").Methods:The sensitivity and accuracy of the reagent were evaluated in accordance with the WHO HDV RNA international standard, employing the Bio-Rad CFX Opus 96 real-time fluorescence quantitative PCR analysis system. Serial dilutions of pseudo-viruses or cell culture-derived virus were used to determine the linear range of the domestic HDV RNA reagent. Specificity was assessed using positive samples of HAV, HBV, HCV infection, and HEV national reference materials. Precision was evaluated with samples at both high and low concentrations. In a comparative analysis, 30 HDV IgG positive samples were tested using both the domestic HDV RNA reagent and the RoboGene HDV RNA kit based on the ABI 7500 FAST DX system. The Pearson correlation coefficient (r) was used to examine the correlation between the two reagents.Results:The domestic HDV RNA reagent demonstrated a high sensitivity of up to 6 IU/ml, consistent with that of the comparator reagent. The calibration curve for WHO HDV RNA standards had a slope of -3.286, with an amplification efficiency of 101.6%. The linear detection range spanned from 10 to 10 8 IU/ml for eight HDV genotypes. The domestic HDV RNA reagent exhibited exceptional specificity, without cross-reactivity observed with HAV, HBV, HCV, or HEV. Accuracy assessments at five concentration levels met the required standards, with intra-assay precision coefficient of variation ( CV) ranging from 1.20% to 4.20%, and inter-assay precision CV from 1.20% to 7.90%. The detection results for HDV IgG positive samples were highly correlated with the comparator reagent ( r=0.984, P<0.001), achieving a diagnostic accuracy of 100% compared to sequencing results. Conclusion:In this study, the domestic HDV RNA reagent possesses excellent specificity, accuracy, precision, and a broad linear range, attaining a sensitivity level on par with international reagents of the same type.

Aiming at the background of budget management reform in public hospitals and the problems existing in the implemen-tation process,it discusses how to strengthen procurement budget management from the perspective of operation management,im-prove the efficiency and effect of budget funds,reduce costs and increase efficiency,prevent risks,and promote high-quality development.It is suggested that efforts should be made to connect procurement budget management with the top-level design of public hospital development,explore the coordination between fund budget and project process management,unify rigid budget constraints with dynamic adjustment,and establish a multi-dimensional,whole-process procurement budget performance evaluation system.

Objective:To investigate the differences in multi-b-value apparent diffusion coefficient (ADC) and exponential apparent diffusion coefficient (eADC) between hepatocellular carcinoma (HCC) and paracancerous liver tissue, distant cancerous liver tissue, and background liver tissues by ultra-high field 3.0T diffusion-weighted (DWI) MRI imaging.Methods:Sixty-eight consecutive HCC cases confirmed by surgical pathology from January 2018 to October 2021 were enrolled and divided into a cirrhosis ( n=39) and a non-cirrhosis group ( n=29) according to the presence or absence of cirrhosis.The average ADC and eADC of liver tissues of paracancerous (including proximal and distal), distant cancerous, and background were measured by DWI images with diffusion sensitivity factors (b) of 50, 100, 400, 600 s/mm 2, and 1 000 s/mm 2, respectively. The Kruskal-Wallis H test and Bonferroni method were used to test the differences between the measured values of the five tissues. The statistical differences were used to evaluate the diagnostic efficacy of the five tissues by parametric receiver operating characteristic (ROC) curve and area under the curve (AUC). Results:The comparison of average ADC and eADC among five types of tissues in the liver cirrhosis group showed that the average ADC and eADC measured at b values of 50, 100, 400, and 600 s/mm 2 had statistically significant differences (adjusted P<0.005) between cancerous and proximal paracancerous, distal paracancerous, distant cancerous, and background liver tissue, as well as the average ADC measured at b=1 000 s/mm 2 between cancerous and proximal paracancerous tissue. The average ADC and eADC in the non-cirrhosis group had statistically significant differences (adjusted P<0.005) between cancerous and proximal paracancerous, distant paracancerous, distant cancerous, and background liver tissue measured at b values of 50, 100, and 400 s/mm 2, respectively. The average ADC and eADC measured at b=600 s/mm 2 showed statistically significant differences (adjusted P<0.005) between cancerous and proximal paracancerous, distal paracancerous, and distant cancerous liver tissue, as well as the average ADC measured at b=1 000 s/mm 2 between cancerous and distal paracancerous, and distant cancerous liver tissue. The average ADC and eADC in the cirrhosis group had no statistically significant difference between the proximal paracancerous and the distant cancerous, as well as the background liver tissue measured at b-values of 50, 100, 400, 600, and 1 000 s/mm 2, respectively (adjusted P>0.005), while there were statistically significant differences (adjusted P<0.005) in the average ADC values in the non-cirrhosis group between the proximal paracancerous and the distant paracancerous and background liver tissues at b=50 s/mm 2, as well as the average ADC and eADC values between the proximal paracancerous and the distant liver tissues at b=100 s/mm 2. The average ADC and eADC values measured in the cirrhosis group and non-cirrhosis group had no statistically significant difference between the distant paracancerous, distant cancerous, and background liver tissue (adjusted P>0.005). The efficacy of average ADC and eADC in distinguishing five types of tissues (cancerous and proximal paracancerous, distant paracancerous, distant cancerous, and background liver tissue) showed that in the cirrhosis group, the diagnostic efficacy was best at b=50 s/mm 2. The area under the ROC curve (AUC) of average ADC was 0.815, 0.828, 0.855, and 0.855, respectively, and the AUC of average eADC was 0.815, 0.830, 0.856, and 0.855, respectively. The diagnostic efficacy was best in the non cirrhosis group at b=100 s/mm 2, with average ADC AUCs of 0.787, 0.823, 0.841, and 0.821, and average eADC AUCs of 0.836, 0.874, 0.893, and 0.873, respectively. The AUC of the average ADC in the non-cirrhosis group for distinguishing between proximal paracancerous and distant cancerous liver tissues, as well as proximal paracancerous and background liver tissues, with b=50 s/mm 2, were 0.605 and 0.604, respectively. The average AUC of ADC and eADC for distinguishing between proximal paracancerous and distant liver tissues with b=100 s/mm 2 were 0.619 and 0.620, respectively. Conclusion:The average ADC and eADC measured by multiple b-values are helpful in distinguishing HCC from proximal paracancerous, distal paracancerous, distant-cancerous, and background liver tissues in patients with cirrhosis and non-cirrhosis, while the average ADC and eADC at b=50 s/mm 2 and 100 s/mm 2 exhibit differences between the proximal paracancerous from the distant cancerous liver tissue and background liver tissue in patients with non-cirrhosis.

Objective To compare the safety and clinical efficacy of freehand and 3D printing navigation template assisted screw placement in patients with old odontoid fractures of type Ⅱ.Methods Total of 38 patients with old odontoid fractures of type Ⅱ were treated from November 2018 to December 2022,all of which presented as chronic neck pain.According to the dif-ferent methods of screw insertion into the pedicle,the patients were divided into a navigation template group and a freehand group.In the navigation template group,there were 17 patients including 9 males and 8 females with an average age of(51.30±13.20)years old,disease duration was(22.18±7.59)months.In the freehand group,there 21 patients including 7 males and 14 females with an average age of(49.46±11.92)years old,disease duration was(19.52±9.17)months.The intraoperative blood loss,operation time,and postoperative drainage output were recorded and compared between two groups.The accuracy of screw placement was evaluated by CT scan.Before operation and 1 year after operation,cervical pain was assessed by visual analogue scale(VAS),neurological changes were evaluated by the Japanese Orthopaedic Association(JOA)score,and the de-gree of spinal cord injury was assessed by the American Spinal Injury Association(ASIA)injury scale.Results All patients were followed up for(25.31±1.21)months.The operation time of template group(112.00±20.48)min had significantly shorter than that of the freehand group(124.29±15.24)min(P＜0.05),while there were no significant differences between two groups in terms of intraoperative blood loss,postoperative drainage,and hospital stay(P＞0.05).At 1 year after operation,in template group and freehand group,the VAS[(2.88±0.86),(2.90±0.83)]and JOA[(14.94±1.82),(14.62±2.19)]improved with pre-operative[VAS(4.71±0.92),(4.86±0.79)and JOA(12.18±2.30),(11.95±2.31)](P＜0.05),with no significant difference between two groups(P＞0.05).No significant improvement was observed in ASIA grading in either group at 1 year after opera-tion(P＞0.05),and there was no significant difference between two groups(P＞0.05).The template group had significantly better accuracy of screw placement in the pedicle of the axis than the freehand group(P＜0.05),while no significant difference was observed between two groups in the accuracy of screw placement in the pedicle of the atlas(P＞0.05).Conclusion In the treat-ment of type Ⅱ old odontoid fractures with posterior pedicle screw fixation,3D printing navigation template screw placement can significantly shorten the operation time,achieve similar clinical efficacy as free-hand screw placement,and significantly im-prove the accuracy of screw placement in the pedicle of the axis.

Humans ; Cardiologists ; Neoplasms/drug therapy* ; Medical Oncology ; Heart ; Oncologists

ObjectivesTo assess the correlation between blastocyst morphology score， serum human chorionic gonadotropin β subunit （β-hCG） levels on day 12 after transfer and live birth outcomes among cycles tested HCG-positive after thawed single blastocyst transfer； to analyze the predictive value of serum β-hCG levels on live birth. MethodsWe reviewed the data of 519 frozen-thawed single blastocyst transfer cycles （FET） that had been tested HCG-positive from January 2016 to May 2020 at our IVF center. These FET cycles were firstly divided into 4 groups （AA， AB， BA， and BB） according to Gardner's grading system of inner cell mass （ICM） and trophectoderm cell （TE）， and then 4 groups （stages 3， 4， 5 and 6） according to the degree of blastocyst expansion. Serum β-hCG concentrations on day 12 after transfer and live birth rates were compared among groups transferred with different blastocysts grading and expansion stage. The relationship between Gardner’s grading or expansion stage of blastocysts and serum β-hCG levels was determined by correlation test， and ROC curves were plotted to determine the threshold values of serum β-hCG for predicting live birth. Results（1） The serum β-hCG concentration in the AA group and AB group on the 12th day after the transfer was significantly higher than that in the BB group （P <0.001， P <0.001）. However， there was no significant difference in the live birth rate when different ICM/TE-graded blastocysts were transferred （P = 0.120）. There were no significant differences in serum β-hCG concentration on day 12 after transfer and live birth rate among blastocysts with different expansion stages （P = 0.091， P = 0.557）. （2） There was a significant weak correlation between blastocyst ICM/TE grading and serum β-hCG concentration on day 12 （r_s = -0.221， P <0.001）， and even after controlling for confounding factors （ r_s = -0.228， P <0.001）；There was no significant correlation between blastocyst’s expansion stage and serum β-hCG concentration on day 12 after the transfer （r_s = -0.052， P = 0.240）， and the association remained insignificant after controlling for confounding factors （r_s = -0.029， P = 0.508）. （3） ROC curve analysis showed that the cut-off value for predicting live birth by serum β-hCG on day 12 was 657.5 mU/mL （P < 0.001）. ConclusionsNeither the ICM/TE grade nor the expansion stage of blastocysts affect the live birth rate，there is significant difference in the level of β-hCG produced by blastocyst with different ICM/TE grade；Our results suggest that early serum β-hCG level can predict live birth.

Recent advances in genome editing, especially CRISPR-Cas nucleases, have revolutionized both laboratory research and clinical therapeutics. CRISPR-Cas nucleases, together with the DNA damage repair pathway in cells, enable both genetic diversification by classical non-homologous end joining (c-NHEJ) and precise genome modification by homology-based repair (HBR). Genome editing in zygotes is a convenient way to edit the germline, paving the way for animal disease model generation, as well as human embryo genome editing therapy for some life-threatening and incurable diseases. HBR efficiency is highly dependent on the DNA donor that is utilized as a repair template. Here, we review recent progress in improving CRISPR-Cas nuclease-induced HBR in mammalian embryos by designing a suitable DNA donor. Moreover, we want to provide a guide for producing animal disease models and correcting genetic mutations through CRISPR-Cas nuclease-induced HBR in mammalian embryos. Finally, we discuss recent developments in precise genome-modification technology based on the CRISPR-Cas system.
Animals ; CRISPR-Cas Systems/genetics* ; DNA/genetics* ; Embryo, Mammalian/metabolism* ; Endonucleases/metabolism* ; Gene Editing ; Mammals/metabolism*

Borderline resectable pancreatic ductal adenocarcinoma accounts for approximately 20% of newly diagnosed pancreatic cancer patients. This type of adenocarcinoma is between resectable and unresectable. It has a high degree of heterogeneity and features in anatomy, biology, and physical condition. The biological characteristics of invasiveness determine that, rather than direct surgery, neoadjuvant therapy should be primarily given to patients to achieve R0 resection and avoid early postoperative recurrence. However, this treatment model is still controversial. According to the latest research on this topic, the full text summarizes the definition of BR-PDAC, resectable evaluation, neoadjuvant treatment selection and evaluation, surgical results after neoadjuvant therapy, and the efficacy of adjuvant therapy after neoadjuvant therapy.