Vertebral fracture severity assessment on anteroposterior radiographs with a new semi-quantitative technique.

We developed a new tool to assess the severity of osteoporotic vertebral fracture using radiographs of the spine. Our technique can be used in patient care by helping to stratify patients with osteoporotic vertebral fractures into appropriate treatment pathways. It can also be used for research purposes. PURPOSE: The aim of our study was to propose a semi-quantitative (SQ) grading scheme for osteoporotic vertebral fracture (OVF) on anteroposterior (AP) radiographs. METHODS: On AP radiographs, the vertebrae are divided into right and left halves, which are graded (A) vertical rectangle, (B) square, (C) traverse rectangle, and (D) trapezoid; whole vertebrae are graded (E) transverse band or (F) bow-tie. Type A and B were compared with normal and Genant SQ grade 1 OVF, Type C and D with grade 2 OVF, and Type E and F with grade 3 OVF. Spine AP radiographs and lateral radiographs of 50 females were assessed by AP radiographs SQ grading. After training, an experienced board-certified radiologist and a radiology trainee assessed the 50 AP radiographs. RESULTS: The height-to-width ratio of the half vertebrae varied 1.32-1.48. On lateral radiographs, 84 vertebrae of the 50 patients had OVFs (38 grade 1, 24 grade 2, and 22 grade 3). On AP radiographs, the radiologist correctly assigned 84.2%, 91.7%, and 77.2% and the trainee correctly assigned 68.4%, 79.2%, and 81.8% of grade 1, 2, and 3 OVFs, respectively. Compared with lateral radiographs, the radiologist had a weighted Kappa of 0.944 including normal vertebrae and 0.883 not including normal vertebrae, while the corresponding Kappa values for the trainee were 0.891 and 0.830, respectively. CONCLUSION: We propose a new semi-quantitative grading system for vertebral fracture severity assessment on AP spine radiographs.

[A case of fungal keratitis caused by Petriella setifera infection].

The patient, a 66-year-old male, suffered from redness, blurred vision, photophobia, and tearing in the right eye after being injured by a wooden board. Anti-inflammatory treatment showed poor effectiveness. A 4 mm × 4 mm infiltrate with white deposits on the surface was observed in the central cornea of the right eye. Microscopic examination of corneal scrapings, fungal culture, and in vivo confocal microscopy all indicated fungal infection. The isolated strain was identified as Scedosporium apiospermum through microscopic morphology and confirmed as Petriella setifera by gene sequencing. The patient received corneal debridement combined with routine anti-inflammatory and antifungal treatment in the outpatient clinic. During the follow-up period, the condition continued to improve. Slit lamp examination at the revisit 40 days after the initial diagnosis revealed thinning of the corneal stroma, basic healing of the epithelium, and an increase in uncorrected visual acuity from 0.3 to 0.6.

[Clinical analysis of lung adenocarcinoma with epidermal growth factor receptor mutation transformed into sarcoma].

Objective: To analyze the clinical data of a case of lung adenocarcinoma with Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) resistance transforming into sarcoma, and to conduct a literature review to improve the understanding of the resistance mechanism. Histological transformation is a unique form of acquired resistance of EGFR-TKIs in non-small cell lung cancer (NSCLC). Thereinto, the transformation of small cell carcinoma is more common, and the transformation of sarcoma is rarely reported. Methods: Clinicopathological data on the treatment process, pathological features, and clinical outcome of the patient with EGFR-TKIs-resistance lung adenocarcinoma transforming into sarcoma were collected. The literature was reviewed to analyze the pathogenetic mechanism for sarcomatoid carcinoma or sarcoma transformation after drug resistance of adenocarcinoma, as well as the clinical characteristics of the patients and the corresponding therapeutic schemes. Results: We reported a patient with lung adenocarcinoma who developed EGFR-T790M mutation after first-line treatment with icotinib and sarcoma transformation after second-line treatment with almonertinib. Chemotherapy, radioactive particle implantation, antiangiogenic therapy and immunotherapy were followed, but the results were unsatisfactory. There was no report of EGFR-TKIs-resistant lung adenocarcinoma transforming into sarcoma. Among the 14 reports of adenocarcinoma transforming into sarcomatoid carcinoma, 8 cases had EGFR mutation, 3 cases had ALK mutation, 2 cases had ROS1 mutation, and 1 case had no asscoiated sensitive mutation. The median survival of 14 patients with adenocarcinoma transforming to sarcomatoid carcinoma was only 3 months. Conclusions: Sarcoma transformation can be one of the forms of drug resistance in patients with lung adenocarcinoma with EGFR-TKIs. The prognosis of patients with adenocarcinoma after transformation into sarcoma is poor.

[Long-term follow-up of humanized and murine CD19 CAR-T-cell therapy for B-cell acute lymphoblastic leukemia].

Objective: Murine CD19 chimeric antigen receptor T-cell (CAR-T) products have been approved for the treatment of refractory/relapsed (R/R) B-cell acute lymphocytic leukemia (B-ALL) ; moreover, humanized products are also undergoing clinical trials. This study aimed to explore the differences in safety and short- and long-term follow-up efficacy between humanized and murine CD19 CAR-T-cells for treating relapsed and refractory B-ALL. Methods: Clinical data of 80 patients with R/R B-ALL treated with CD19-targeted CAR-T-cells at the Union Hospital of Tongji Medical College of Huazhong University of Science and Technology between May 2016 and March 2023 were analyzed, which included 31 patients with murine CAR-T and 49 with humanized products. Results: The proportion of patients with cytokine-release syndrome (CRS) in the murine and humanized groups was 63.1% and 65.3%, respectively. Moreover, a higher proportion of patients suffered from severe CRS in the murine group than in the humanized CAR-T group (19.4% vs 8.2%, P=0.174). Furthermore, one patient per group died of grade 5 CRS. The incidence of grade 1-2 immune effector cell-associated neurotoxicity syndrome (ICANS) was 12.9% and 6.1%, respectively; severe ICANS were not observed. Among patients receiving murine CAR-T-cells, an overall response (OR) was observed in 74.2%. Conversely, the OR rate of patients receiving humanized CAR-T-cells was 87.8%. During the median follow-up time of 10.5 months, the median recurrence-free survival (RFS) of patients with murine CAR-T-cells was 12 months, which was as long as that of patients with humanized CAR-T-cells. The median overall survival (OS) were not reached in both groups. Of the 45 patients with a bone marrow burden over 20% at baseline, humanized CAR-T therapy was associated with a significantly improved RFS (43.25% vs 33.33%, P=0.027). Bridging transplantation was an independent factor in prolonging OS (χ(2)=8.017, P=0.005) and PFS (χ(2)=6.584, P=0.010). Common risk factors, such as age, high proportion of bone marrow blasts, and BCR-ABL fusion gene expression, had no significant effect on patients' long-term follow-up outcomes. Three patients reached complete remission after reinfusion of humanized CAR-T-cells. However, one patient relapsed one month after his second infusion of murine CAR-T-cells. Conclusions: The results indicate that humanized CAR-T therapy showed durable efficacy in patients with a higher tumor burden in the bone marrow without any influence on safety. Moreover, it could overcome immunogenicity-induced CAR-T resistance, providing treatment options for patients who were not treated successfully with CAR-T therapies.

[Pathological study on the relationship between nucleic acid oxidative stress and heart failure with preserved ejection fraction in patients aged over 85 years].

Objective: To investigate the level of nucleic acid oxidation in myocardial tissue of patients aged over 85 with heart failure with preserved ejection fraction (HFpEF) and the correlation with myocardial amyloid deposition. Methods: This was a retrospective case-control study. Data of patients≥85 years old who underwent systematic pathological autopsy in Beijing Hospital from 2003 to 2017 were retrospectively collected. Twenty-six patients were included in the HFpEF group and 13 age-and sex-matched patients who had not been diagnosed with heart failure and died of non-cardiovascular diseases served as the control group. The left ventricular myocardium slices of both groups were semi-quantitatively analyzed using immunohistochemical staining of 8-oxidized guanine riboside (8-oxo-G) and 8-oxidized guanine deoxyriboside (8-oxo-dG) to evaluate the oxidation of RNA and DNA in cardiomyocytes. Using the median of the mean absorbance value of 8-oxo-G immunohistochemical staining as the cut-off value, patients were divided into high-absorbance group and low-absorbance group. Congo red staining was used to compare myocardial amyloid deposition between the two groups. Results: The mean age of patients in HFpEF group was (91.8±3.7) years, 24 (92.3%) were males. The mean age of patients in control group was (91.7±3.7) years old, 11 (84.6%) were males. The median mean optical absorbance value of 8-oxo-G immunohistochemical staining of myocardium was significantly higher in HFpEF patients than in control group (0.313 8 (0.302 2, 0.340 6) vs. 0.289 2 (0.276 7, 0.299 4), Z=-3.245, P=0.001). The median mean absorbance value of 8-oxo-dG immunohistochemical staining of myocardial tissue was similar between the two groups (0.300 0 (0.290 0, 0.322 5) vs. 0.300 0 (0.290 0, 0.320 0), Z=-0.454, P=0.661). Proportion of patients with moderate and severe cardiac amyloid deposition was significantly higher in the high-absorbance group than in the low-absorbance group ((85.0%, 17/20) vs. (31.6%, 6/19), P=0.001). Conclusion: The RNA oxidation degree of myocardium in HFpEF patients is higher than that in elderly people without heart failure. Degree of myocardial amyloid deposits is higher in patients with high levels of RNA oxidation.

Genomic epidemiological investigation of an outbreak of Serratia marcescens neurosurgical site infections associated with contaminated haircutting toolkits in a hospital barber shop.

BACKGROUND: Nine surgical site infections caused by Serratia marcescens were diagnosed in neurosurgical patients in a 3500-bed hospital between 2nd February and 6th April 2022. OBJECTIVE: To trace the source of infections caused by S. marcescens to expedite termination of the outbreak and prevent future epidemics. METHODS: A review of all surgical procedures and cultures yielding S. marcescens since February 2022 was conducted. Samples were collected from patients and environmental sources. S. marcescens isolates were characterized by antibiotic susceptibility testing. Whole-genome sequencing (WGS) was used to investigate genetic relationships. Resistance genes, virulence genes and plasmid replicons were identified. RESULTS: S. marcescens was isolated from patients' puncture fluid, cerebrospinal fluid and other secretions, and was also cultured from the barbers' haircutting tools, including leather knives, slicker scrapers and razors. In total, 15 isolates were obtained from patients and eight isolates were obtained from haircutting tools. All isolates exhibited identical antibiotic resistance patterns. WGS revealed close clustering among the 23 isolates which differed significantly from previous strains. Three resistance genes and nine virulence-associated genes were detected in all isolates, and 19 of 23 isolates harboured an MOBP-type plasmid. The results confirmed an outbreak of S. marcescens, which was traced to contaminated haircutting tools in the hospital barber shop. The outbreak ended after extensive reinforcement of infection control procedures and re-education of the barbers. CONCLUSIONS: These results highlight the risk of postoperative infections related to pre-operative skin preparation, and demonstrate the value of next-generation sequencing tools to expedite outbreak investigations.

[Clinicopathological features of esophageal carcinoma with ductal differentiation of esophageal gland].

Objective: To investigate the clinicopathological characteristics of esophageal carcinoma with gland duct differentiation. Methods: The clinical, morphologic and immunohistochemical (IHC) features of eight cases of esophageal carcinoma with gland duct differentiation diagnosed from 2012 to 2022 at the First Affiliated Hospital of Soochow University were summarized. Results: There were four males and four females, with a mean age of 68.5 (range 59-82) years. Two tumors were located in middle esophagus, five in the lower esophagus, and one in the cardia. The mean diameter was 2.4 cm (range 0.6-4.5 cm). The tumor had a bilayer epithelial structure, including the inner luminal epithelium and the outer basal epithelium. Immunohistochemistry showed that CK7 (8/8) and CK18 (8/8) were positive in the inner epithelium. p40 (8/8), p63 (8/8) and CK5/6 (8/8) were positive in the outer epithelium. SMA, calponin and CD117 were all negative. p53 mutants were found in all eight cases (strong and diffuse positivity in 6/8; complete loss of expression in 2/8). No columnar metaplasia, intestinal metaplasia and ectopic gastric mucosa were observed in the surface squamous epithelium in the cases. The mean follow-up time was 21.5 months (range 5-51 months). Seven patients survived and one patient died 31 months after surgery due to recurrence and liver metastasis. Conclusion: Esophageal carcinoma with esophageal gland duct differentiation is a rare tumor with unique histologic and IHC characteristics.

[The effect of chronic endometritis on the clinical outcomes of patients with failure of first embryo transfer].

Objective: To investigate the effect of chronic endometritis (CE) on the clinical outcomes of patients with failure of first embryo transfer. Methods: A total of 5 605 cycles of frozen-thawed single blastocyst transfer in the reproductive center of the Third Affiliated Hospital of Zhengzhou University from January 2017 to June 2021 were retrospectively collected. After the failure of first embryo transfer, all patients underwent hysteroscopy, and when necessary, endometrial pathology and immunohistochemistry were combined to diagnose CE. Patients were divided into two groups: non-CE group (5 033 cycles) and CE treatment group (572 cycles). The main outcome was live birth rate and the secondary outcomes included clinical pregnancy rate and early abortion rate. The quantitative data were represented by Median (Q1, Q3). The rank sum test was used for comparison between groups. The factors related to live birth rate were analyzed by binary logistic regression model. Results: The incidence of CE was 10.21% (572 cycles) in patients with the failure of first embryo transfer. The maternal age in the non-CE group was 31.0 (29.0, 34.0) years old, and that in the CE treatment group was 31.0 (29.0, 34.0) years old (P<0.001). There was a statistically significant difference in endometrial preparation between the two groups (P=0.010). The endometrial thickness in the CE group was 9.0 (8.2, 10.3) mm on progesterone transformation day, which was higher than that of [9.5 (8.6, 11.0) mm] in the non-CE group (P<0.001). There was no significant difference in clinical pregnancy rate (60.3% (3 035 cycles) vs 63.1% (361 cycles), P=0.193), early abortion rate (17.1% (520 cycles) vs 20.5% (74 cycles), P=0.112) and live birth rate (49.2% (2 477 cycles) vs 49.3% (282 cycles), P=0.969) between the non-CE group and the CE treatment group. The maternal age, endometrial thickness on progesterone transformation day and blastocyst grade were related factors of the live birth rate, and the OR(95%CI) were 0.94 (0.93-0.96), 1.10 (1.06-1.14) and 2.07 (1.84-2.32)), respectively (all P<0.001). Compared with the non-CE group, the CE treatment group did not affect the live birth rate after transplantation, the aOR (95%CI) was 0.99 (0.82-1.18), P=0.882. Conclusions: For patients who underwent the failure of first embryo transfer, hysteroscopy is recommended before single frozen blastocyst transfer, and if necessary, combined with immunohistochemical screening for CE. After standardized treatment, CE patients could obtain similar clinical pregnancy rate, early miscarriage rate and live birth rate as non-CE patients.

Nutrient supplementation of beef female calves at pre-weaning enhances the commitment of fibro-adipogenic progenitor cells to preadipocytes.

We aimed to evaluate the impact of nutrient supplementation of beef female calves at pre-weaning on adipogenic determination. Thirty-four female calves were assigned to two experimental treatments: Control (CON, n = 17), where animals were supplemented only with mineral mixture; Supplemented (SUP, n = 17), where animals received energy-protein supplement containing minerals (5 g/kg of BW per day) of their body weight. Animals were supplemented from 100 to 250 days of age, and muscle samples were biopsied at the end of the supplementation period. Regarding the performance variables, there were no differences between treatments for initial body weight (P = 0.75). The final body weight (P = 0.07), average daily gain (P = 0.07), rib eye area (P = 0.03), and rib fat thickness (P = 0.08) were greater in SUP female calves compared with CON treatment. The number of fibro-adipogenic progenitor cells (P = 0.69) did not differ between treatments, while a greater number of intramuscular pre-adipocytes were observed in SUP than CON female calves (P = 0.01). The expression of miRNA-4429 (P = 0.20) did not differ between treatments, while the expression of miRNA-129-5p (P = 0.09) and miRNA-129-2-3p (P = 0.05) was greater in CON than SUP female calves. Our results suggest that nutrient supplementation at early postnatal stages of development enhances the commitment of fibro-adipogenic progenitor cells into the adipogenic lineages allowing to an increase in intramuscular fat deposition potential of the animals later in life.

Swinging between shine and shadow: Theoretical advances on thermally activated vibropolaritonic chemistry.

Polariton chemistry has emerged as an appealing branch of synthetic chemistry that promises mode selectivity and a cleaner approach to kinetic control. Of particular interest are the numerous experiments in which reactivity has been modified by virtue of performing the reaction inside infrared optical microcavities in the absence of optical pumping; this effort is known as "vibropolaritonic chemistry." The optimal conditions for these observations are (1) resonance between cavity and reactive modes at normal incidence (k = 0) and (2) a monotonic increase of the effect with the concentration of emitters in the sample. Importantly, vibropolaritonic chemistry has only been experimentally demonstrated in the so-called "collective" strong coupling regime, where there is a macroscopic number of molecules (rather than a single molecule) coupled to each photon mode of the microcavity. Strikingly, efforts to understand this phenomenon from a conceptual standpoint have encountered several roadblocks, and no single, unifying theory has surfaced thus far. This Perspective documents the most relevant approaches taken by theorists, laying out the contributions and unresolved challenges from each work. We expect this Perspective to not only serve as a primer for experimentalists and theorists alike but also inform future endeavors in the quest for the ultimate formalism of vibropolaritonic chemical kinetics.

Does Periodontitis Affect the Association of Biological Aging with Mortality?

The prevalence of periodontitis is increasing with the aging of the global population. Periodontitis has been suggested to accelerate aging and increase mortality. The present nationwide prospective cohort study aimed to determine whether periodontitis could modify the association of biological aging with all-cause and cause-specific mortality in middle-aged and older adults. Participants ≥40 y of age from the Third National Health and Nutrition Examination Survey (NHANES III) were included (n = 6,272). Phenotypic age acceleration (PhenoAgeAccel) was used to evaluate the biological aging process. Moderate/severe periodontitis was defined using a half-reduced Centers for Disease Control and Prevention and American Academy of Periodontology case definition. Multivariable Cox proportional hazard regression was conducted to estimate the association between PhenoAgeAccel and mortality risk, followed by effect modification analysis to test whether periodontitis modified the association. During a median follow-up of 24.5 y, 3,600 (57.4%) deaths occurred. The positive relationships between PhenoAgeAccel and all-cause and cause-specific mortality were nonlinear. After adjusting for potential confounders, the highest quartile of PhenoAgeAccel was associated with increased all-cause mortality in individuals with no/mild periodontitis (hazard ratio for Q4 vs. Q1 [HRQ4vs.Q1] = 1.789; 95% confidence interval [CI], 1.541-2.076). In contrast, the association was enhanced in patients with moderate/severe periodontitis (HRQ4vs.Q1 = 2.446 [2.100-2.850]). Periodontal status significantly modified the association between PhenoAgeAccel and all-cause mortality (P for interaction = 0.012). In subgroup analyses, the modifying effect of periodontitis was observed in middle-aged adults (40-59 y), females, and non-Hispanic Whites. Although cause-specific mortality showed a similar trend, the PhenoAgeAccel × periodontitis interaction did not reach statistical significance. In conclusion, periodontitis might enhance the association of biological aging with all-cause mortality in middle-aged and older adults. Hence, maintaining and enhancing periodontal health is expected to become an intervention to slow aging and extend life span.

Analysis of Brachypodium distachyon UVR8 reveals conservation in UV-B receptors.

The Ultraviolet Resistance Locus 8 (UVR8) in plants recognizes ultraviolet-B (UV-B) light and plays a crucial role in regulating plant growth through a series of signal transduction events. However, the UVR8 in monocotyledon crops has not yet been systematically analysed. We identified BdUVR8 (BRADI_3g45740) from the genome of Brachypodium distachyon, a relative of wheat, by analysing the phylogenetic tree, the gene expression pattern, detecting accumulation of UV-B response metabolites, and checking for phenotype recovery. The BdUVR8 protein sequence is similar to the known UVR8 of other species. The phylogenetic tree of UVR8 shows clear divergence between dicotyledons and monocotyledons. Expression analysis revealed that UV-B downregulates BdUVR8 by 70% and upregulates the chalcone synthase (BdCHS) gene 3.4-fold in B. distachyon. The pCAMBIA1300::BdUVR8-mCherry construct introduced into Arabidopsis uvr8 mutants showed that the BdUVR8 protein is localized in the cytoplasm and translocates into the nucleus in response to UV-B irradiation. The introduction of BdUVR8 into uvr8 rescued hypocotyl elongation caused by UV-B and restored expression of HY5, Chalcone synthase, and Flavanone 3-hydroxylase, as well as accumulation of total flavonoids. Together, our results show that BdUVR8 is a photoreceptor that perceives UV-B in B. distachyon.

[Analysis of vaginal microecology in 23 181 cases of the gynecological female outpatients].

Objective: To analyze the vaginal microecological status of vaginitis population and non-vaginitis population of gynecological female outpatients. Methods: A total of 30 265 women who visited the gynecological outpatient clinic of Beijing Obstetrics and Gynecology Hospital from December 2018 to December 2020 completed vaginal microecological examination. After removing the follow-up patients, 23 181 women were divided into group with symptoms and signs of vaginitis (6 697 cases) and group without symptoms and signs of vaginitis (16 484 cases), according to whether the women with symptoms and signs of vaginitis or not. And the vaginal microecological status of the two groups was compared and analyzed. Results: (1) The total detection rate of vaginitis in the initial women was 34.87% (8 083/23 181), of which 46.10% (3 087/6 697) in group with symptoms and signs of vaginitis and 30.31% (4 996/16 484) in group without symptoms and signs of vaginitis, nearly 1/3 of the gynecological outpatients without signs and symptoms of vaginitis had vaginitis. (2) Among the types of simple vaginitis, vulvovaginal candidiasis (VVC) was the most frequent in group with symptoms and signs of vaginitis (16.01%, 1 072/6 697), followed by aerobic vaginitis (AV; 12.83%, 859/6 697), with significant differences compared with group without symptoms and signs of vaginitis (all P<0.001). There were no statistical differences between the two groups of bacterial vaginosis (BV) and trichomonal vaginitis (TV), indicating that BV and TV were more likely to be neglected (all P>0.05). (3) The proportion of various combinations of vaginitis among 2 632 cases of mixed vaginitis were, in descending order: BV+AV, VVC+AV, BV+AV+VVC, AV+TV, AV+TV+BV, BV+VVC. (4) Microecological analysis of 15 098 cases diagnosed with non-vaginitis had normal flora (including those with normal flora and those with normal flora but decreased function) in 14 013 cases (92.81%, 14 013/15 098), abnormal flora in 429 cases (2.84%, 429/15 098) and the BV intermediate in 656 cases (4.34%, 656/15 098); this indicated that the vast majority of the microecological tests were normal in the vaginal microbiota of those without vaginitis. Conclusions: Microecological examination could diagnose multiple pathogenic infections at once, and is especially important as a guide for the definitive diagnosis of mixed vaginitis and vaginitis with atypical clinical symptoms. Vaginal infections such as BV and TV that are easily overlooked should be concerned.

[The expression and function of PD-L1 in CD133(+) human liver cancer stem-like cells].

Objective: To investigate the expression of programmed death protein-ligand 1 (PD-L1) in liver cancer stem-like cells (LCSLC) and its effect on the characteristics of tumor stem cells and tumor biological function, to explore the upstream signaling pathway regulating PD-L1 expression in LCSLC and the downstream molecular mechanism of PD-L1 regulating stem cell characteristics, also tumor biological functions. Methods: HepG2 was cultured by sphere-formating method to obtain LCSLC. The expressions of CD133 and other stemness markers were detected by flow cytometry, western blot and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect the expressions of stemness markers and PD-L1. The biological functions of the LCSLC were tested by cell function assays, to confirm that the LCSLC has the characteristics of tumor stem cells. LCSLC was treated with cell signaling pathway inhibitors to identify relevant upstream signaling pathways mediating PD-L1 expression changes. The expression of PD-L1 in LCSLC was down regulated by small interfering RNA (siRNA), the expression of stem cell markers, tumor biological functions of LCSLC, and the changes of cell signaling pathways were detected. Results: Compared with HepG2 cells, the expression rate of CD133 in LCSLC was upregulated [(92.78±6.91)% and (1.40±1.77)%, P<0.001], the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were also higher than those in HepG2 cells (P<0.05), the number of sphere-formating cells increased on day 7 [(395.30±54.05) and (124.70±19.30), P=0.001], cell migration rate increased [(35.41±6.78)% and (10.89±4.34)%, P=0.006], the number of transmembrane cells increased [(75.77±10.85) and (20.00±7.94), P=0.002], the number of cloned cells increased [(120.00±29.51) and (62.67±16.77), P=0.043]. Cell cycle experiments showed that LCSLC had significantly more cells in the G(0)/G(1) phase than those in HepG2 [(54.89±3.27) and (32.36±1.50), P<0.001]. The tumor formation experiment of mice showed that the weight of transplanted tumor in LCSLC group was (1.32±0.17)g, the volume is (1 779.0±200.2) mm(3), were higher than those of HepG2 cell [(0.31±0.06)g and (645.6±154.9)mm(3), P<0.001]. The expression level of PD-L1 protein in LCSLC was 1.88±0.52 and mRNA expression level was 2.53±0.62, both of which were higher than those of HepG2 cells (P<0.05). The expression levels of phosphorylation signal transduction and transcription activation factor 3 (p-STAT3) and p-Akt in LCSLC were higher than those in HepG2 cells (P<0.05). After the expression of p-STAT3 and p-Akt was down-regulated by inhibitor treatment, the expression of PD-L1 was also down-regulated (P<0.05). In contrast, the expression level of phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) in LCSLC was lower than that in HepG2 cells (P<0.01), there was no significant change in PD-L1 expression after down-regulated by inhibitor treatment (P>0.05). After the expression of PD-L1 was knockdown by siRNA, the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were decreased compared with those of siRNA-negative control (NC) group (P<0.05). The number of sphere-formating cells decreased [(45.33±12.01) and (282.00±29.21), P<0.001], the cell migration rate was lower than that in siRNA-NC group [(20.86±2.74)% and (46.73±15.43)%, P=0.046], the number of transmembrane cells decreased [(39.67±1.53) and (102.70±11.59), P=0.001], the number of cloned cells decreased [(57.67±14.57) and (120.70±15.04), P=0.007], the number of cells in G(0)/G(1) phase decreased [(37.68±2.51) and (57.27±0.92), P<0.001], the number of cells in S phase was more than that in siRNA-NC group [(30.78±0.52) and (15.52±0.83), P<0.001]. Tumor formation in mice showed that the tumor weight of shRNA-PD-L1 group was (0.47±0.12)g, the volume is (761.3±221.4)mm(3), were lower than those of shRNA-NC group [(1.57±0.45)g and (1 829.0±218.3)mm(3), P<0.001]. Meanwhile, the expression levels of p-STAT3 and p-Akt in siRNA-PD-L1 group were decreased (P<0.05), while the expression levels of p-ERK1/2 and ß-catenin did not change significantly (P>0.05). Conclusion: Elevated PD-L1 expression in CD133(+) LCSLC is crucial to maintain stemness and promotes the tumor biological function of LCSLC.

Neoadjuvant chemoradiotherapy versus neoadjuvant chemotherapy followed by minimally invasive esophagectomy for locally advanced esophageal squamous cell carcinoma: a prospective multicenter randomized clinical trial.

BACKGROUND: Neoadjuvant therapy is recommended for locally advanced esophageal cancer, but the optimal strategy remains unclear. We aimed to evaluate the safety and efficacy of neoadjuvant chemoradiotherapy (nCRT) versus neoadjuvant chemotherapy (nCT) followed by minimally invasive esophagectomy (MIE) for locally advanced esophageal squamous cell carcinoma (ESCC). PATIENTS AND METHODS: Eligible patients staged as cT3-4aN0-1M0 ESCC were randomly assigned (1 : 1) to the nCRT or nCT group stratified by age, cN stage, and centers. The chemotherapy, based on paclitaxel and cisplatin, was administered to both groups, while concurrent radiotherapy was added for the nCRT group; then MIE was carried out. The primary endpoint was 3-year overall survival. This study is registered with ClinicalTrials.gov (NCT03001596). RESULTS: A total of 264 patients were eligible for the intention-to-treat analysis. By 30 November 2021, 121 deaths had occurred. The median follow-up was 43.9 months (interquartile range 36.6-49.3 months). The overall survival in the intention-to-treat population was comparable between the nCRT and nCT strategies [hazard ratio (HR) 0.82, 95% confidence interval (CI) 0.58-1.18; P = 0.28], with a 3-year survival rate of 64.1% (95% CI 56.4% to 72.9%) versus 54.9% (95% CI 47.0% to 64.2%), respectively. There were also no differences in progression-free survival (HR 0.83, 95% CI 0.59-1.16; P = 0.27) and recurrence-free survival (HR 1.07, 95% CI 0.71-1.60; P = 0.75), although the pathological complete response in the nCRT group (31/112, 27.7%) was significantly higher than that in the nCT group (3/104, 2.9%; P < 0.001). Besides, a trend of lower risk of recurrence was observed in the nCRT group (P = 0.063), while the recurrence pattern was similar (P = 0.802). CONCLUSIONS: NCRT followed by MIE was not associated with significantly better overall survival than nCT among patients with cT3-4aN0-1M0 ESCC. The results underscore the pending issue of the best strategy of neoadjuvant therapy for locally advanced bulky ESCC.

Surgical treatment of peri-implantitis with or without adjunctive graft material: a systematic review and meta-analysis of randomized controlled trials.

This systematic review and meta-analysis was performed to compare the clinical effect of surgical treatment of peri-implantitis alone or in combination with graft material. Literature searches were conducted up to June 20, 2020. Randomized controlled trials (RCTs) comparing the clinical effects of open flap debridement (OFD) alone and OFD with adjunctive graft materials for the treatment of peri-implantitis were included. Probing depth (PD) changes and marginal bone level (MBL) changes were assessed and expressed as the standardized mean difference (SMD) with 95% confidence interval (CI). Subgroup analyses and sensitivity analyses were conducted. The search yielded 7419 articles, five of which were analysed quantitatively. The adjunctive use of graft materials in OFD did not provide additional PD improvements (SMD 0.46, 95% CI -0.13 to 1.05; P = 0.13), but did yield additional MBL improvements (SMD 1.04, 95% CI 0.71-1.37; P < 0.01). The degradability of the material, number of implants included per patient, and risk of bias did not have significant effects on the results, but the origin of the material may affect the PD improvements. Based on the available evidence, the adjunctive use of graft materials in the surgical treatment of peri-implantitis can significantly improve MBL changes but not PD changes.

Tumor necrosis factor alpha and palmitate simulate bovine fatty liver disease in vitro when using abattoir-derived primary bovine hepatocytes isolated by a novel nonperfusion method.

Hepatic lipidosis (i.e., fatty liver) is a common periparturient disease in high-producing dairy cattle affecting nearly 50% of cows to some degree and costing an estimated 60 million dollars annually. Large animal studies are costly, labor intensive, and are not well suited to mechanistic studies. Traditionally, mechanistic studies employ in vitro methodologies, utilizing established cell lines or primary cell culture methods. However, with dairy cattle, established hepatic cell lines do not exist, and methods for primary cell culture studies typically involve complicated procedures that often utilize very young animals (typically bull calves). Several previously published papers have used abattoir-derived tissues as a source of primary cells; however, a simple method utilizing simple culture media has yet to be presented. In addition, we sought to develop a way to replicate the syndrome of fatty liver disease "in a dish" using adult cattle that should more closely represent the physiology of the periparturient dairy cow. Herein we present a non-perfusion-based method that results in robust growth and proliferation of abattoir-derived bovine hepatocytes that demonstrate lipid loading, elevated lactate dehydrogenase leakage, and cytotoxicity as demonstrated by elevated caspase 3/7 expression consistent with in vivo physiology of the periparturient dairy cow with fatty liver disease.