Objective:To investigate the effect of melatonin (MEL) on learning and memory abilities of fluoride-exposed offspring rats and the role of gut microbiota.Methods:Twelve 8-week-old Sprague-Dawley (SD) rats (8 females and 4 males) with a body weight ranging from 180 to 220 g were selected and divided into control group 1 and fluoride-exposed group 1 using a random number table method, with 6 rats in each group (female ∶ male = 2 ∶ 1). They were free to drink purified water or purified water containing 100 mg/L sodium fluoride, respectively. After 2 months, male and female rats were raised together in cages, and the first postnatal day (PND) of the offspring rats was recorded as PND0. In PND21, the offspring rats of fluoride-exposed group 1 were divided into fluoride-exposed group (Group F, n = 6) and fluoride + MEL group (Group FM, n = 6) using a group design, and continued to be exposed to fluoride through drinking water. The offspring rats of control group 1 were divided into control group (Group C, n = 6) and MEL group ( n = 6). The groups FM and MEL were given 20 mg/kg MEL by gavage, while the groups C and F were given the same dose of normal saline by gavage. In PND60, novel object recognition and Morris water maze tests were used to observe the learning and memory abilities of the offspring rats. Western blotting (WB) was used to detect the expression level of brain derived neurotrophic factor (BDNF) in the hippocampus of the offspring rats. And 16S rDNA sequencing technology was used to detect the changes in the structure and composition of gut microbiota in fecal samples. Results:The results of novel object recognition test showed that there was a statistically significant difference in the discrimination index (DI) among the four groups of offspring rats ( F = 3.95, P = 0.024). The DI in groups C and FM was higher than that of Group F ( P < 0.05). The results of Morris water maze test showed that compared with Group C, the platform-crossing time of the offspring rats of Group F were less and they had a longer time to reach the platform for the first time ( P < 0.05). Compared with Group F, the platform-crossing time of the offspring rats of Group FM were increased and they had a shorter time to reach the platform for the first time ( P < 0.05). The WB results showed that compared with Group C (1.00 ± 0.07), the expression level of BDNF protein in Group F (0.68 ± 0.26) was lower ( P < 0.05). Compared with Group F, the expression level of BDNF protein in Group FM (0.99 ± 0.14) was higher ( P < 0.05). Anosim similarity analysis showed significant differences in the structure and composition of gut microbiota in the four groups of offspring rats ( R = 0.395 062, P = 0.002). The distribution characteristics of gut microbiota species showed that at the phylum level, compared with Group C, the relative abundance of Bacteroidetes in Group F increased from 14.26% to 37.00%, and the relative abundance of Firmicutes decreased from 68.78% to 45.95%. Compared with Group F, the relative abundance of Firmicutes in Group FM increased from 45.95% to 65.26%, and the relative abundance of Bacteroidetes decreased from 37.00% to 23.00%. At the genus level, compared with Group C, the relative abundance of Lactobacillus, Dubosiella, HT002 and UCG-005 in Group F was lower, while the relative abundance of unclassified Muribaculaceae was higher. Compared with Group F, the relative abundance of Lactobacillus, Dubosiella, HT002 and UCG-005 in Group FM was higher, while the relative abundance of unclassified Muribaculaceae was lower. The results of linear discriminant analysis revealed that the Candidatus-Saccharimonas and Incertae-Sedis were significantly enriched in Group C, unclassified Muribaculaceae and Muribaculum were significantly enriched in Group F, and Allorhizobium- Neorhizobium- Pararhizobium- Rhizobium were significantly enriched in Group FM. Conclusion:MEL can improve the learning and memory impairment of offspring rats induced by fluoride exposure by changing the structure and composition of gut microbiota.

Objective To investigate the potential of immune-inflammatory markers and the characteristics of magnetically controlled capsule endoscopy in distinguishing gastric adenocarcinoma from precancerous lesions,as well as to develop and validate a risk prediction model.Methods Retrospective analysis was conducted on medical records of 578 patients who underwent magnetic controlled capsule endoscopy at our hospital between January 2021 and December 2023.Following the principle of Pareto's law(80/20 rule),they were randomly divided into a training set(462 cases)and a validation set(116 cases).Magnetic controlled capsule endoscopy and blood cell tests were performed,with pathological diagnosis results serving as the"gold standard",to classify patients into groups of gastric adenocarcinoma and precancerous lesions.The magnetic controlled capsule endoscopic features,neutrophil-lymphocyte ratio(NLR),platelet-lymphocyte ratio(PLR)in patients with gastric adenocarcinoma and precancerous lesions were compared to develop and validate a risk diagnostic model for gastric adenocarcinoma.Results Among the 462 patients who underwent magnetic controlled capsule endoscopy,gastric adenocarcinoma was diagnosed in 76 cases through pathological examination,accounting for 16.45%(76/462),while precancerous lesions were observed in 386 cases,accounting for 83.55%(386/462).In the validation set of 116 patients who underwent gastric endoscopy,there were 22 cases of gastric adenocarcinoma,representing an incidence rate of 18.97%(22/116),and a total of 94 cases with precancerous lesions,accounting for an incidence rate of 81.03%(94/116).No statistically significant differences(P＞0.05)were found between the two groups regarding lesion size,border appearance,mucus presence or lesion morphology.However,compared to the precancerous lesion group,the proportion of whitish coloration as well as irregular surface microstructure and grid-like microvessels was significantly higher in the gastric adenocarcinoma group(P＜0.05).Moreover,both NLR and PLR values were significantly higher in the gastric adenocarcinoma group compared to those in the precancerous lesion group(P＜0.05).Irregular surface microstructure(OR=2.213,95%CI:1.288～3.801),irregular grid-like microvessels(OR=2.489,95%CI:1.458～4.249),NLR(OR=2.369,95%CI:1.389～4.046),and PLR(OR=3.016,95%CI:1.767～5.148)were identified as risk factors for gastric adenocarcinoma(P＜0.05).The sensitivity of the risk model for diagnosing gastric adenocarci-noma in the training set was 0.800(95%CI:0.716～0.891),with a specificity of 0.783(95%CI:0.694～0.851)and an area under the curve of 0.858(95%CI:0.787～0.931).In the validation set,the sensitivity for diagnosing gastric adenocarcinoma was 0.861(95%CI:0.771～0.945),with a specificity of 0.769(95%CI:0.683～0.841)and an area under the curve of 0.844(95%CI:0.765～0.923).Conclusion The surface microstructure,microvas-cular morphology,NLR,and PLR of gastric lesions are correlated with the occurrence of gastric adenocarcinoma.Developing a risk diagnostic model facilitates early identification and diagnosis of gastric adenocarcinoma.

ObjectiveTo introduce the Hr gene of spontaneously mutated SHJH^hr mice into BALB/cAShjh inbred mice with clear genetic background,and provide a basis for study on the molecular mechanism of Hr gene mutation-induced abnormal phenotype and the application of this model.Methods Using a backcross-intercross breeding method guided by phenotypic monitoring, mutant genes from SHJH^hr mice bred by spontaneous mutation were introduced into inbred BALB/cAShjh mice by homozygous mutation introgression, and the mice were bred into BALB/cA.Cg.SHJH^hr (abbreviated as C.Cg.SHJH^hr) mice after 10 generations. The genotypes of 90 single nucleotide polymorphism (SNP) detection sites were analyzed in C.Cg.SHJH^hr mice by multiplex PCR library construction followed by next generation sequencing. Then 14 biochemical locus marker genes were detected in C.Cg.SHJH^hr mice according to the method of GB/T 14927.1-2008. Finally, whole genome exon sequencing was utilized to detect the mutated genes in this mouse. ResultsFrom May 2018 to March 2022, a total of 10 generations of backcross-intercross were conducted to complete the construction of the C.Cg.SHJH^hr mouse line. Among the 90 SNPs loci detected, except for rs13484115 and rs13484116, all the other loci had the same genotype as the recipient mice BALB/cAShjh. The results of biochemical marker gene detection showed that all the 14 loci of the mouse were the same as those of the recipient mouse. Whole genome exon sequencing found that the mouse had 109 site mutations compared with the recipient mouse strain, including 71 synonymous mutations, 1 stopgain, 37 missense mutations, and 20 genes involved in protein sequence alterations (including the reported Hr gene). ConclusionC.Cg.SHJH^hr mice were created. Through exon sequencing and genetic analysis, three Hr mutated genes and associated mutated genes that mainly cause phenotypic variations were identified, which provides a basis for expanding the application of C.Cg.SHJH^hr mice in biomedical research.

Objective To measure and analyze biological characteristics and aging phenotype of SHJH^hr mice and provide basic data for the application of the mouse model in aging mechanisms research and antiaging drug development. MethodsWith ICR mice of the same age as control group, the body mass growth data of SHJH^hr mice at the age of 3 to 16 weeks, the reproduction ability of 1 to 4 fetuses and the life cycle of SHJH^hr mice were measured. Blood routine (30 items) and serum biochemical indexes (25 items) of 6-week-old SHJH^hr mice were measured. The venous blood of 8-week-old SHJH^hr mice was collected for flow cytometry analysis to determine the content of immune cells. The aging bone structure of the cancellous bone and bone mineral density of SHJH^hr mice aged 4, 8 and 26 weeks were measured by micro-CT. Histopathological changes of bone and joint of 8-week-old mice were observed. ResultsCompared with ICR mice, the female and male body mass of SHJH^hr mice were significantly lower at the age of 16 weeks (P < 0.05), and the reproductive performance of female mice was low (P < 0.01) or did not have normal reproductive capacity. The shortest survival time of SHJH^hr mice was 57 weeks and the longest was 71 weeks, which was shorter than those of normal ICR mice, showing obvious rapid aging phenomenon. At the same time, some physiological and biochemical indexes of blood and pathological changes of bone and cartilage tissues also showed the accelerated aging and abnormality of animal physiological functions. ConclusionSHJH^hr mice have some biological characteristics of rapid aging as well as some physiological and pathological changes caused by aging.

【Objective】 To evaluate the effect of one-beat acquisition with wide detector CT on the image quality and diagnostic efficiency of coronary CT angiography (CCTA) in patients with atrial fibrillation. 【Methods】 A total of 52 consecutive patients with atrial fibrillation, including 31 males, (67.32±11.45) years old, who underwent CCTA from July 2022 to February 2023, were analyzed retrospectively. All patients underwent one-beat acquisition CCTA. The subjective and objective image quality of the coronary arteries was evaluated, and using invasive coronary catheter angiography as the gold standard, the diagnostic efficacy of stenosis degrees above moderate and severe degrees was calculated, respectively. 【Results】 Subjective evaluation results: 92.31% (384/416) of the vascular segments were rated as excellent or good, and the diagnosable rate reached 98.08% (408/416, subjective score ≥3 points). Objective evaluation results: The CT value of the right coronary artery, anterior descending branch, and circumflex branch was (433.41±95.17)HU, (422.69±92.81)HU and (420.27±95.43)HU, respectively; the contrast-to-noise ratio was 38.46±7.54, 32.46±13.78 and 37.74±8.89, respectively. The total diagnostic accuracy, sensitivity, and specificity was 94.71%, 87.9% and 96.62%, respectively, for moderate stenosis and 96.15%, 83.64% and 98.06% for severe stenosis. 【Conclusion】 One-beat acquisition with wide detector CT can obtain high-quality coronary artery images and high diagnostic accuracy for patients with atrial fibrillation without radiation dose increase to patients. It has good clinical application value for patients with atrial fibrillation.

【Objective】 To construct the eukaryotic expression vector carrying the human wild-type p27 and lacking nuclear localization signal p27△NLS coding sequences， and the express them in HEK293T cells， which may contribute to investigating the different locations and roles of p27 in the cytoplasm and nucleus. 【Methods】 Total RNA was prepared from human breast cancer MCF7 cells， and cDNA was obtained by reverse transcription-polymerase chain reaction （RT-PCR）. After amplification of the p27 CDs and non-NLS fragments by PCR， full length p27WT （CDKN1B， NM_004064.5） and p27△NLS coding regions were obtained. PCR products were then subcloned into the eukaryotic expression vector pCMV-Blank. After identification with bacterial PCR， double restriction enzyme digestion and sequencing， they were defined officially as pCMV-p27WT and pCMV-p27△NLS， respectively. The recombinant plasmids were transfected into HEK293T cells by electroporation. After 48 h， the levels of p27 protein in the cytoplasm and nucleus were detected by Western blotting. 【Results】 The sequencing results showed that the sequences of p27WT and p27△NLS inserted into the plasmids were both correctly consistent with that of NM_004064.5. After transfection with pCMV-p27WT， total p27 protein expression was increased and distributed in both the cytoplasm and nucleus of HEK293T cells. After transfection with pCMV-p27△NLS， p27 protein was significantly increased and almost entirely localized in the cytoplasm of HEK293T cells. 【Conclusion】 The eukaryotic expression plasmids of human p27WT and p27△NLS coding sequences were successfully constructed and overexpressed in HEK293T cells. This research may lay a foundation for investigating the biological function of p27 in the cell cycle progression of tumor cells.

Objective:To investigate the role of survival motor neuron ( SMN) gene knockout in mice with cisplatin-induced acute kidney injury (AKI). Methods:A mouse model (C57BL/6) of cisplatin-induced AKI was constructed. Twenty male wild type (WT) and SMN+/- mice weighing 22-24 g were randomly divided into four groups: WT mice with saline injection group (WT vehicle, n=5), SMN+/- mice with saline injection group ( SMN+/- vehicle, n=5), WT mice with cisplatin injection group (WT cisplatin, n=5) and SMN+/- mice with cisplatin injection group ( SMN+/- cisplatin, n=5). Mice were injected intraperitoneally with 20 mg/kg cisplatin or 0.9% saline. 72 hours later, the mice were sacrificed, and serum and kidney tissues were collected. The real time PCR and Western blotting were used to measure the expression levels of SMN mRNA and protein. The sarcosine oxidation and urease method were used to measure serum creatinine (Scr) and blood urea nitrogen (BUN) levels. Renal pathologic changes were observed by PAS staining. TUNEL immunofluorescence assay was used to detect the level of apoptosis. Western blotting and immunohistochemistry were used to detect the protein expression levels of apoptosis index poly (ADP-ribose) polymerase (PARP) and endoplasmic reticulum stress index CHOP. Results:Compared with WT mice, SMN mRNA and protein expression levels were lower in SMN+/- mice, and the expression level of SMN mRNA and protein was further decreased after intraperitoneal cisplatin injection (all P<0.05). Compared with WT mice with saline injection group, WT mice with cisplatin injection group had higher levels of Scr, BUN, tubular damage scores, TUNEL positive cell numbers, PARP and CHOP, while the expression levels of above indexes in the SMN+/- mice with cisplatin injection group were higher than those in the WT mice with cisplatin injection group (all P<0.05). Conclusions:SMN gene knockout can aggravate renal pathological damage and apoptosis of renal tubular epithelial cell in cisplatin-induced AKI mice. SMN may be a potential therapeutic target of AKI.

Chronic heart failure is a serious heart disease with dyspnea and limited activity tolerance as the main clinical manifestations. Activation of the inflammatory system can significantly stimulate cardiac fibrosis and remodeling and promote the progression of heart failure， playing a key role in the development of the disease. Studies have confirmed that inflammation is involved in the development of different types of heart failure. "Toxic pathogen theory" is an important basic theory of traditional Chinese medicine （TCM） to explain the occurrence of diseases. We concluded the similarities between TCM toxic pathogens and inflammation in concept， disease location， etiology， syndrome differentiation， and clinical characteristics. Chronic heart failure is manifested by the toxic pathogens of turbid phlegm， stagnated blood， and accumulated fluid. Heart vessel obstruction is the main pathological factor， and the inflammatory factors produced by necrotic cardiomyocytes are the microscopic manifestations of the obstruction. Therefore， based on the "toxic pathogen theory"， this study aimed to clarify the role of inflammation in the development of chronic heart failure from both macroscopic and microscopic perspectives. Moreover， this paper proposed that the stagnated blood has not been transformed into toxin in the early stage of the disease and thus the products of clearing heat and detoxification should not be used. At the development stage of the disease when the transformation tends to begin， treatment should be based on syndrome differentiation， and detoxifying Chinese medicine should be used in order to achieve the goal of "removing toxin without harming the healthy Qi". At the late stage of heart failure， toxins have been accumulated and detoxifying medicines and therapies should be applied to eliminate the toxic pathogens. This study is expected to lay a foundation for the modern research on the role of inflammation in the development of chronic heart failure with TCM theory and guide the diagnosis and treatment of this disease.

【Objective】 To compare the enhancement effects of lean body weight （LBW） and total body weight （TBW） as indexes to calculate the contrast agent dosage under the condition of energy spectrum CT scanning. 【Methods】 A total of 218 patients who received liver enhancement CT from November 2018 to January 2019 were enrolled in this study. There were 101 patients in LBW group and 117 patients in TBW group. Both groups were scanned by energy spectrum CT， and the parameters of scanning and reconstruction were identical. The contrast agent dose was 500 mgI/kg （LBW） in LBW group and 450 mgI/kg （TBW） in TBW group， and the injection rate was 2.8 mL/s. Images were transferred to a GE AW4.7 workstation and the 50 keV monochromatic images were analyzed. We compared the dosage of contrast medium， CT value of aorta in arterial phase （HU-aorta）， hepatic enhancement CT value in venous phase （-liver）， the rate of reaching the enhancement standard and variability in the two groups. 【Results】 Compared with TBW group， LBW group had lower contrast agent dosage， HU-aorta and ∆-liver （P<0.05）， LBW and TBW group had no statistically different enhanced rate of HU-aorta as （91.09% vs. 90.60%） or ∆-liver （92.08% vs. 88.89%） （P>0.05）. The variation rate of HU-aorta and ∆-liver in LBW group was lower than that in TBW group. Using LBW as an index to calculate the dosage of liver enhanced CT also made the enhancement of liver parenchyma more consistent in different patients. 【Conclusion】 Even on the premise of energy spectrum CT scanning， using LBW-based contrast injection in liver enhanced CT can not only reduce contrast dose， but also make the enhancement in liver parenchyma more consistent among different patients.

【Objective】 To investigate the effects of 80 kVp combined with multi-model adaptive statistical iterative reconstruction algorithm （ASiR-V） on the image quality of CT angiography （CTA） of iliac artery in kidney transplantation candidates before operation. 【Methods】 Totally 50 kidney transplantation candidates underwent “one-stop” scanning combined CTA for coronary and iliac arteries were recruited consecutively. After the scanning， images were reconstructed with different ASiR-V levels on the iliac artery from 50% to 100% at a 10% interval， and 6 groups of images were obtained. We evaluated and compared all image qualities， DLP and CTDI in the patients’ examination were recorded， and the effective dose （ED） was calculated. 【Results】 All of the image quality objective scores of different ASiR-V levels on the iliac artery were above 3 points （3.32±0.24）， and the average CT value was （344.30±74.53）HU. The ED received by the patient throughout the examination was （2.71±0.42)mSv. The image noise decreased while SNR and CNR increased monotonically as the ASiR-V levels increased， among which 80% ASiR-V image quality score was the highest with 3.41±0.26. 【Conclusion】 For patients who need to have both coronary and iliac arteries evaluated before kidney transplantation operation， low tube voltage （80 kVp） combined with 80% ASiR-V can obtain high-quality iliac artery images under the premise of reducing the ED dose， which can provide practical basis for further reducing the dose in personalized scanning scheme for such patients. It has good feasibility and clinical application value.