ObjectiveTo establish background data for a 90-day feeding trial of SD rats to ensure the reliability of research data. MethodsBackground data from six independent 90-day feeding trials of SD rats conducted by the National Center for Safety Evaluation of Drugs from 2020 to 2023 were summarized. These studies involved a blank control group of 120 SPF-grade 4-week-old SD rats, with an equal number of males and females, which were only given standard full-nutrient pelleted rat feed. After the quarantine period, the animals were observed for an additional 90 days, followed by intraperitoneal injection of Zoletil (50 mg/mL) for anesthesia, blood sampling, euthanasia, and necropsy. By analyzing the data from the blank control group, a relevant background database for SD rats was established. ResultsBoth male and female rats exhibited steady weight gain, with a more pronounced increase in male rats. Within 90 days, the average body weight of male and female rats increased to over 500 g and 300 g, respectively. Three weeks later, the average daily food intake of male rats stabilized at approximately 25~28 g per rat, while that of female rats remained stable at approximately 16~19 g per rat. The food utilization rate of all animals gradually decreased from the first week of the experiment. In the white blood cell (WBC) differential count results, significant differences were observed in the counts of WBCs, neutrophils (Neut), lymphocytes (Lymph), and monocytes (Mono) between males and females (P<0.001). However, there were no significant differences in the percentages of neutrophil (%Neut), lymphocyte (%Lymph), and monocyte (%Mono) between the sexes (P>0.05). The average red blood cell count (RBC), hemoglobin concentration (HGB), hematocrit (HCT), platelet count (PLT), prothrombin time (PT), and activated partial thromboplastin time (APTT) were higher in male animals than in female animals (P<0.05). The average values of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine phosphokinase (CK), lactate dehydrogenase (LDH), glucose (GLU), and triglyceride (TG) in male rats were higher than those in female rats (P<0.05). The urinary pH range for male animals was 5.0 to 8.5, while for female animals it was 6.5 to 9.0. The majority of male animals had a urinary specific gravity lower than 1.020, and the majority of female animals had a urinary specific gravity lower than 1.015. The weights of various organs (excluding the adrenal glands and reproductive organs) in male animals were heavier than those in female animals (P<0.001), while the organ/body weight ratios (excluding the kidneys and reproductive organs) of female animals were higher than those of male animals (P<0.001). ConclusionThis study summarizes the background reference ranges for body weight, food intake, hematology, and serum biochemistry indicators in SPF-grade SD rats in the untreated control group from six 90-day feeding trials conducted by the National Center for Safety Evaluation of Drugs. It provides important reference data for related research. By summarizing the background and spontaneous histopathological changes in rats, this study aids in the standardization and normalization of subsequent research, as well as in the evaluation and analysis of abnormal results.

Objective To investigate the current situation of Staphylococcus aureus contamination in raw and cooked meat and dairy products in Wuhan, and analyze the enterotoxins production and antimicrobial resistance of isolated bacterial strains. Methods The detection of Staphylococcus aureus was performed according to GB4789.10-2016 National Food Safety Standard. Staphylococcus aureus enterotoxin (SE) PCR kit and ELISA were used for SEA-E type detection. Broth dilution and PCR method were used for drug sensitivity test. Results A total of 13 strains of Staphylococcus aureus were isolated from 202 samples, and the isolation rate of Staphylococcus aureus in the raw and cooked meat and dairy products was 6.43%. The detection rate of Staphylococcus aureus was 9.82% (11/112) in raw meat, and 4% (2/50) in cooked meat products. There was no detection in dairy products. Of the 13 isolated strains, 6 strains were found to have enterotoxins, with a toxin production rate of 46.15% (6/13). Among the 6 strains of enterotoxin producing Staphylococcus aureus, 4 strains were classified as type A, C, D, and AB, respectively. The isolated strains were generally resistant to tetracycline and sulfonamide drugs, and the detection rate of resistant genes was more than 60%. The resistance rate to penicillin and erythromycin exceeded 50%, and the dominant resistance spectrum was the detection of 3 strains of single-resistant (PEN) Staphylococcus aureus (25.08%, 3/13), followed by 2 strains of five-fold resistance (PEN-ERY-CLI-SXT-GEN), and double resistance Staphylococcus aureus (PEN-ERY) (15.38%, 2/13). Genetic testing was consistent with phenotypic testing. Conclusion In 2020, there was a certain degree of contamination of Staphylococcus aureus in raw and cooked meat products in Wuhan, with 13 isolated strains and 6 strains producing enterotoxins. It is necessary to remain vigilant about the potential food risks of raw and cooked meat products, and strengthen the supervision of the safety risks of raw and cooked meat products.

Objective:To analyze the effect of the implementation of diagnosis-intervention packet (DIP) on the doctors′ diagnosis and treatment behavior of chronic diseases, so as to provide reference for further improving medical insurance payment related policies.Methods:The first page information of chronic disease patients admitted to hospitals with diabetes, hypertension and coronary atherosclerotic heart disease as the main conditions in 103 hospitals at all levels and township health centers in a city from 2016 to 2020 was collected, and the patients were divided into non-DIP group and DIP group according to the implementation time of DIP. After 1∶1 propensity score matching to balance the general conditions of the 2 groups, the diagnosis and treatment behaviors were analyzed from two dimensions: diagnostic behavior and treatment behavior. The grade A rate of medical record writing, admission and discharge diagnosis coincidence rate, and the average length of stay were used to evaluate the diagnostic behavior; the proportion of drugs and the degree of change in the cost structure were used as the evaluation indicators of treatment behavior.Results:After matching, 41 050 patients were included in both the non-DIP group and the DIP group.From the perspective of diagnostic behavior, the grade A rate of medical record writing in the non-DIP group and the DIP group was 99.40% and 99.83%, the coincidence rate of admission and discharge diagnosis was 58.42% and 61.79%, the average hospital stay was 8.03 days and 7.04 days respectively, and the difference between the groups was significant ( P<0.05). From the view of treatment behavior, the proportion of drugs decreased from 33.00% in the non-DIP group to 27.59% in the DIP group, with a significant difference ( P<0.05); the drug cost represented by Western medicine changed negatively, while the diagnostic cost showed a positive change. Conclusions:DIP has played a certain role in regulating doctors′ diagnosis and treatment behavior for chronic diseases. Among them, doctors have significantly improved their diagnostic behavior for chronic diseases, and the proportion of drugs in treatment behavior has been well controlled.

Objective:To explore the value of echo-planar imaging correction (EPIC) for improving image quality of diffusion weighted imaging (DWI) and diffusion tensor imaging (DTI) of cervical cord.Methods:A total of 33 subjects (20 males, 13 females) were scanned on a 3.0 T MR scanner from January to March 2022, and the sequences included T 1WI, DWI and DTI (with and without corrections). Two observers delineated the regions of interest (ROIs) on the fused images of DWI and DTI with T 1WI before and after correction, and measured the average diffusion coefficient (ADC), fractional anisotropy (FA), and offset distance of ROIs between images with and without corrections. The subjective scores of image quality were also evaluated. The ICC or Kappa was used to test the consistency of the quantitative measurement and subjective scores by the two observers. The average values by the two observers would be used for subsequent analysis. The independent pair t-test and Wilcoxon test were used for comparison of objective measurements and Mann-Whitney U test was used for subjective image assessments between images with and without corrections. Results:The measurement data and the subjective scores of the two observers were in good agreement (ICC 0.912-0.999, Kappa 0.778-0.816). The independent sample t-test showed the subjective scores were significantly different for the DWI and DTI images between before and after geometry and/or ADC corrections. The ADC values of C6, the offset distances measured by DWI before and after correction of C4, C5, and C6 and subjective scores were significantly different ( P<0.05); The FA values of C1 and C3, ADC values of C1 and C3, offset distance of C4, C5 and C6 measured by DTI before and after correction and subjective scores were statistically significant ( P<0.001). Conclusion:EPI geometry correction and ADC value correction can significantly reduce geometric distortion, increase image quality, and thus improve the diagnosis accuracy of essential diseases.

Objective:To evaluate rapid on-site evaluation (ROSE) in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) for pancreatic solid lesions, and to compare the difference in ROSE performance between cytopathologists and trained endoscopists.Methods:A total of 168 consecutive patients with pancreatic solid lesions who underwent EUS-FNA from January 2014 to December 2020 at Fuding Hospital, Fujian University of Traditional Chinese Medicine were recruited. The patients who did not receive ROSE from January 2014 to November 2017 were included in N-ROSE group ( n=67). Since December 2017, the patients who intended to receive EUS-FNA were divided into E-ROSE group ( n=59, patients who received EUS-FNA and ROSE by endoscopists trained with cytopathology) and C-ROSE group ( n=42,patients who received EUS-FNA by untrained endoscopists and ROSE by cytopathologists) according to random number table. The number of punctures, sample adequacy, cytological diagnosis, final diagnosis and diagnostic efficiency (including the sensitivity, the specificity, the positive predictive value, the negative predictive value and the accuracy) in 3 groups were compared. Results:(1) The puncture number in N-ROSE group (4.22±0.76) was significantly more than E-ROSE group (3.12±0.79, P<0.001) and C-ROSE group (3.24±0.91, P<0.001). (2) The proportions of adequate samples in N-ROSE group [82.09% (55/67)] was significantly lower than those of E-ROSE group [96.61% (57/59), χ2=5.308, P=0.021] and C-ROSE group [97.62% (41/42), χ2=4.541, P=0.033]. The proportion of negative cytological diagnosis in N-ROSE group [40.30% (27/67)] was significantly higher than those of E-ROSE group [20.34% (12/59), χ2=5.848, P=0.016] and C-ROSE group [19.05% (8/42), χ2=5.348, P=0.021]. (3) The sensitivity of N-ROSE group [74.07% (40/54)] was significantly lower than those of E-ROSE group [94.00% (47/50), χ2=6.151, P=0.013] and C-ROSE group [94.44% (34/36), χ2=4.817, P=0.028]. The accuracy in N-ROSE group [79.10% (53/67)] was significantly lower than those of E-ROSE group [94.92% (56/59), χ2=5.433, P=0.020] and C-ROSE group [95.24% (40/42), χ2=4.155, P=0.042]. (4) There was no significant difference in any observational index between E-ROSE group and C-ROSE group ( P>0.05). Conclusion:ROSE in EUS-FNA can improve sample adequacy, the diagnostic sensitivity and accuracy, and reduce the number of punctures. The sample adequacy and diagnostic efficiency of endoscopists trained with cytopathology are comparable to those of cytopathologists.

Objective:To study the effects of naringenin on pancreatic fibrosis in the mouse model of chronic pancreatitis (CP) and its effects on the activation, proliferation and apoptosis of pancreatic stellate cells (PSCs).Methods:Eighteen C57BL/6 mice were randomly divided into control group, CP group and naringenin group, with 6 mice in each group. The CP mouse model was established by intraperitoneal injections of caerulein. Naringenin group was given naringenin (200 mg/kg/day) by gavage once a day from the first day of the fourth week of modeling process to the day before the killing; the control group and CP group were treated by gavage with an equivalent amount of drug solvent containing 0.5% sodium carboxymethyl cellulose (CMC-Na). Mice were killed 5 days after the last caerulein injection, and their pancreatic tissues were collected for hematoxylin-eosin staining and Sirius Red staining, pathological scoring and collagen sedimentation detection. Naringenin with different concentrations (0, 5, 10, 20, 50, 100, 150, 200 μmol/L) were used to intervene HPSC for 24 hours, and CCK-8 method was used to detect the cell activity. TGF-β1 recombinant protein (2 ng/ml) was used to induce PSCs for 1 hour (TGF-β1 stimulation group), and naringenin with low (50 μmol/L), middle (100 μmol/L) and high (150 μmol/L) concentration was used to intervene for 36 hours after TGF-β1 stimulation, respectively. Western Blotting was used to detect the expression of PSC activation related proteins FN and COL1A1, cell proliferation marker p21, anti-apoptotic protein Bcl-xL, pro-apoptotic protein Bax and Bid.Results:The pathological scores of pancreatic tissue [(7.33±1.15), (4.67±1.15)] and the percentage of collagen positive areas [(46±4), (28±2)%] in CP group and naringenin group were higher than those in the control group [0, (4±2)%]. However, these indexes in the naringenin group were lower than those in CP group, and the differences were all statistically significant (all P value <0.05). The relative expression of FN in control group, TGF-β1 stimulation group and low, medium and high naringenin group was 0.02, 0.76, 0.67, 0.34 and 0.07, respectively; the expression of COL1A1 in these groups was 0.51, 1.71, 1.34, 0.84 and 0.11. The expression of FN and COL1A1 in TGF-β1 stimulation group was significantly higher than that in control group, and the expression of FN and COL1A1 in low, medium and high naringenin group was significantly lower than that in TGF-β1 stimulation group, and the differences were all statistically significant (all P value <0.05). The expression of p21 in the above five groups was 0.87, 1.18, 1.27, 1.22 and 1.00. The expression of p21 in TGF-β1 stimulation group was higher than that in control group, and the expression of p21 in high naringenin group was obviously lower than that in TGF-β1 stimulation group, and the differences were all statistically significant (all P value <0.05). In addition, the expression of Bcl-xL in these groups was 2.09, 2.21, 2.38, 2.50 and 2.12; the expression of Bax was 0.98, 0.88, 0.98, 1.00 and 0.88; the expression of Bid was 1.15, 1.09, 1.14, 1.18 and 1.18. There was no statistically significant difference among these groups (all P value >0.05). Conclusions:Naringenin could significantly alleviate the inflammation, atrophy and fibrosis in the CP mouse model, and inhibit the activation and proliferation of PSCs. However, naringenin had no significant effect on the apoptosis of PSCs, indicating that naringenin may be potentially used to treat pancreatic fibrosis in CP.

Microneedle percutaneous immunization is achieved by puncturing the stratum corneum of the skin with microneedles so that the vaccine is efficiently recognized by antigen-presenting cells to induce a specific immune response. Due to the advantages of efficient induction of immune response, low pain and easy storage, transdermal immunization by microneedles has been widely used for immunization of various vaccines in recent years. This review summarizes the materials of microneedles, application for transcutaneous immunization, as well as the challenges that need to be addressed.
Administration, Cutaneous ; Drug Delivery Systems ; Needles ; Vaccination ; Vaccines

Although widely applied in treating hematopoietic malignancies, transplantation of hematopoietic stem/progenitor cells (HSPCs) is impeded by HSPC shortage. Whether circulating HSPCs (cHSPCs) in steady-state blood could be used as an alternative source remains largely elusive. Here we develop a three-dimensional culture system (3DCS) including arginine, glycine, aspartate, and a series of factors. Fourteen-day culture of peripheral blood mononuclear cells (PBMNCs) in 3DCS led to 125- and 70-fold increase of the frequency and number of CD34⁺ cells. Further, 3DCS-expanded cHSPCs exhibited the similar reconstitution rate compared to CD34⁺ HSPCs in bone marrow. Mechanistically, 3DCS fabricated an immunomodulatory niche, secreting cytokines as TNF to support cHSPC survival and proliferation. Finally, 3DCS could also promote the expansion of cHSPCs in patients who failed in HSPC mobilization. Our 3DCS successfully expands rare cHSPCs, providing an alternative source for the HSPC therapy, particularly for the patients/donors who have failed in HSPC mobilization.
Antigens, CD34/metabolism* ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans ; Leukocytes, Mononuclear/metabolism* ; Peptides/metabolism*

OBJECTIVE: To analyze the results of concurrent hearing and deafness genetic screening and follow up of newborns. METHODS: In total 33 911 babies born to 5 designated hospitals in Nanshan District of Shenzhen city from October 2017 to December 2019 were included. All subjects underwent concurrent hearing and deafness genetic screening covering 21 variants of 4 genes including GJB2, SLC26A4, GJB3 and Mt12SrRNA. For those with positive results, Sanger sequencing was carried out for confirmation. RESULTS: 93.32% subjects passed the first-round hearing screening, and 87.01% passed the recheck testing. The overall detection rate was 4.18%. The detection rates for GJB2, SLC26A4, GJB3 and Mt12srRNA variants were 1.98%, 1.58%, 0.37% and 0.25%, respectively. 126 and 84 subjects were found with high risk for delayed-onset and drug-induced hearing loss, respectively. In addition, 4 and 5 subjects were found to harbor homozygous/compound heterozygous variants of the GJB2 and SLC26A4 genes, respectively. Concurrent screening showed that subjects (with heterozygous variants) who did not passed the two round hearing test were as follows: GJB2 with 6.75% in the first round and 2.61% in the second round testing, SLC26A4 (3.3%/1.2%), GJB3 (0.72%/0.14%) and 12SrRNA (0.36%/Nil), respectively. Moreover, the No-pass rate in the subjects with homozygous or compound variants in single gene, heterozygous variant in single gene, heterozygous variant in multiple genes, and homozygous variant in GJB3 gene were significantly higher than the subjects with negative results of genetic screening. CONCLUSION Concurrent newborn genetic screening can enhance the effectiveness of hearing screening and enable earlier identification and intervention for children with hearing impairment. Follow-up can improve the diagnostic rate for children who are positive for the concurrent screening. Nevertheless, genetic and hearing screening cannot replace the diagnostic testing. It is necessary to conduct comprehensive analysis for the results of genetic and hearing screening and radiological examinations. Sanger sequencing and next-generation sequencing are critical for ascertain the diagnosis.
China/epidemiology* ; DNA Mutational Analysis ; Deafness/genetics* ; Follow-Up Studies ; Genes/genetics* ; Genetic Testing/statistics & numerical data* ; Hearing/genetics* ; Hearing Tests/statistics & numerical data* ; Humans ; Infant, Newborn ; Mutation ; Neonatal Screening

OBJECTIVE: To explore the genetic basis for a Chinese pedigree affected with amyloidosis cutis dyschromica. METHODS: High-throughput sequencing was carried out for the proband. Bioinformatic analysis was used to identify the pathogenic variants. The result was verified by Sanger sequencing. RESULTS: A homozygous nonsense variant c.565C>T (p.Arg189X) of the GPNMB gene was identified in the proband, his elder brother and younger sister, which resulted a truncated protein with loss of function. The father of the proband was a heterozygous carrier for the variant. The genotype of his mother was unknown since she had passed away. Based on the American College of Medical Genetics and Genomics standards and guidelines, the c.565C>T variant was predicted to be likely pathogenic (PS3+ PM2+ PP1+PP3). CONCLUSION The novel homozygous GPNMB variant probably underlay the amyloidosis cutis dyschromica in this pedigree. Above finding has expanded the spectrum of GPNMB gene variants.
Amyloidosis, Familial/genetics* ; China ; Female ; Homozygote ; Humans ; Male ; Membrane Glycoproteins/genetics* ; Mutation ; Pedigree