Objective:To explore the differences in bacterial communities within tissues during the process of oral mucosal carcinogenesis, and analyze the relationship between the high-abundance species Prevotella intermedia (Pi) and the occurrence and development of oral mucosal carcinogenesis. Methods:Fresh tissue samples were collected from patients diagnosed with oral leukoplakia (OLK), oral squamous cell carcinoma (OSCC), and healthy controls (HC) at Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Nanjing University, from January 2022 to November 2024, following strict inclusion criteria. Bacterial DNA was extracted from these specimens, and the 2bRAD sequencing for microbiome (2bRAD-M) was employed to analyze and compare the α and β diversity, as well as the community composition of bacteria within tissues, aiming to identify specifically expressed bacteria. Subsequently, paraffin-embedded clinical specimens were collected: 15 cases in the OLK group (including 4 cases of simple hyperplasia, 6 cases of mild dysplasia, and 5 cases of moderate to severe dysplasia), 12 cases in the OSCC group, and 5 cases in the HC group. A 4-nitroquinoline N-oxide (4NQO)-induced OLK progression mouse model was also constructed. Mice were randomly divided into three groups using a random number table, with six in each group. The negative control group was given distilled water to drink; Group 1 was given distilled water containing 4NQO to drink until week 12, while Group 2 was given distilled water containing 4NQO to drink until week 22. After the mice were sacrificed, their tongue tissue were collected and fixed. Fluorescence in situ hybridization (FISH) with specific probes was used to validate the presence of Pi in human and mouse tissue sections, analyzing the correlation between histopathological grading and the invasion depth of Pi.Results:The 2bRAD-M microbial analysis revealed that the relative abundance of Pi in OSCC tissues (10.80%) was significantly higher than in the HC group (0.50%) ( P=0.001) and OLK group (0.70%) ( P=0.002). FISH probe detection showed that the fluorescence intensity of Pi in human OSCC tissues [123.50 (101.00, 142.30)] was higher than in the HC group [0.00 (0.00, 28.50)], simple hyperplasia OLK [0.00 (0.00, 35.25)], and mild dysplasia OLK [24.50 (0.00, 55.50)] groups, with statistically significant differences respectively ( P=0.002, P=0.003, P=0.005). However, there was no significant difference compared to moderate to severe dysplasia OLK [56.00 (28.00, 62.50)] ( P=0.210). The fluorescence area of Pi in human OSCC tissues [8 615.00 (7 439.00, 11 084.00)] was significantly larger than in the HC group [0.00 (0. 00, 45.00)], simple hyperplasia OLK group [0.00 (0.00, 81.00)], mild dysplasia [49.00 (0.00, 151.00)], and moderate to severe dysplasia groups [1 450.00 (454.00, 2 892.00)], with highly significant differences ( P<0.001). There was a significant correlation between the invasive depth of Pi and the degree of histopathological grading ( P<0.001). In mice, the fluorescence intensity of Pi in OSCC tissues [120.00 (110.00, 127.00)] was significantly higher than in the HC group [0.00 (0.00, 12.25)] ( P<0.01), but showed no significant difference compared with the OLK group [50.00 (0.00, 58.00)] ( P>0.05). The fluorescence area of Pi in mice OSCC tissues [11 020.00 (6 790.00, 12 102.00)] was significantly larger than in the HC group [0.00 (0.00, 56.75)] and the OLK group [0.00 (0.00, 751.50)] ( P=0.006, P=0.043). There is a significant correlation between the depth of invasion of Pi and the degree of histopathological grading ( P<0.01). Conclusions:This study suggests that Pi in oral mucosal tissue may be a potential biomarker for early detection of OSCC and play an important role in the carcinogenesis process of oral mucosa.

Objective:To explore the differences in bacterial communities within tissues during the process of oral mucosal carcinogenesis, and analyze the relationship between the high-abundance species Prevotella intermedia (Pi) and the occurrence and development of oral mucosal carcinogenesis. Methods:Fresh tissue samples were collected from patients diagnosed with oral leukoplakia (OLK), oral squamous cell carcinoma (OSCC), and healthy controls (HC) at Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Nanjing University, from January 2022 to November 2024, following strict inclusion criteria. Bacterial DNA was extracted from these specimens, and the 2bRAD sequencing for microbiome (2bRAD-M) was employed to analyze and compare the α and β diversity, as well as the community composition of bacteria within tissues, aiming to identify specifically expressed bacteria. Subsequently, paraffin-embedded clinical specimens were collected: 15 cases in the OLK group (including 4 cases of simple hyperplasia, 6 cases of mild dysplasia, and 5 cases of moderate to severe dysplasia), 12 cases in the OSCC group, and 5 cases in the HC group. A 4-nitroquinoline N-oxide (4NQO)-induced OLK progression mouse model was also constructed. Mice were randomly divided into three groups using a random number table, with six in each group. The negative control group was given distilled water to drink; Group 1 was given distilled water containing 4NQO to drink until week 12, while Group 2 was given distilled water containing 4NQO to drink until week 22. After the mice were sacrificed, their tongue tissue were collected and fixed. Fluorescence in situ hybridization (FISH) with specific probes was used to validate the presence of Pi in human and mouse tissue sections, analyzing the correlation between histopathological grading and the invasion depth of Pi.Results:The 2bRAD-M microbial analysis revealed that the relative abundance of Pi in OSCC tissues (10.80%) was significantly higher than in the HC group (0.50%) ( P=0.001) and OLK group (0.70%) ( P=0.002). FISH probe detection showed that the fluorescence intensity of Pi in human OSCC tissues [123.50 (101.00, 142.30)] was higher than in the HC group [0.00 (0.00, 28.50)], simple hyperplasia OLK [0.00 (0.00, 35.25)], and mild dysplasia OLK [24.50 (0.00, 55.50)] groups, with statistically significant differences respectively ( P=0.002, P=0.003, P=0.005). However, there was no significant difference compared to moderate to severe dysplasia OLK [56.00 (28.00, 62.50)] ( P=0.210). The fluorescence area of Pi in human OSCC tissues [8 615.00 (7 439.00, 11 084.00)] was significantly larger than in the HC group [0.00 (0. 00, 45.00)], simple hyperplasia OLK group [0.00 (0.00, 81.00)], mild dysplasia [49.00 (0.00, 151.00)], and moderate to severe dysplasia groups [1 450.00 (454.00, 2 892.00)], with highly significant differences ( P<0.001). There was a significant correlation between the invasive depth of Pi and the degree of histopathological grading ( P<0.001). In mice, the fluorescence intensity of Pi in OSCC tissues [120.00 (110.00, 127.00)] was significantly higher than in the HC group [0.00 (0.00, 12.25)] ( P<0.01), but showed no significant difference compared with the OLK group [50.00 (0.00, 58.00)] ( P>0.05). The fluorescence area of Pi in mice OSCC tissues [11 020.00 (6 790.00, 12 102.00)] was significantly larger than in the HC group [0.00 (0.00, 56.75)] and the OLK group [0.00 (0.00, 751.50)] ( P=0.006, P=0.043). There is a significant correlation between the depth of invasion of Pi and the degree of histopathological grading ( P<0.01). Conclusions:This study suggests that Pi in oral mucosal tissue may be a potential biomarker for early detection of OSCC and play an important role in the carcinogenesis process of oral mucosa.

Objective:To analyze the influencing factors and improvement paths of the cultivation of full-time doctoral scientific and technological innovation ability in large public hospitals, and propose countermeasures and suggestions.Methods:This studyed conducted a survey and analysis of 122 doctors from Linyi People′s Hospital in Shandong Province, and completed a current situation study based on the analysis results.Results:There was no significant difference between the two groups in gender, age, degree type, professional category, discipline level, Graduate School type, job type and other indicators. There were significant differences between the two groups in scientific research topic selection ability score, project design ability score, data analysis ability score, data interpretation ability score, project approval in recent 5 years, project level, number of SCI journal papers published in recent 5 years, cumulative impact factors of SCI journal papers, and annual number of academic activities ( P<0.05). Conclusions:The hospital can improve the scientific and technological innovation ability of full-time doctors by setting up a special cultivation plan, establishing an interdisciplinary team, optimizing scientific research management services, improving the evaluation and assessment system, and improving welfare protection.

Objective To investigate the action mechanism of Bufei Yiqi Tongluo formula in treating id-iopathic pulmonary fibrosis (IPF) based on the transforming growth factor-β1 (TGF-β1)/c-Jun N-terminal ki-nase (JNK) signaling pathway.Methods The key targets of network pharmacology conducted the enrichment analysis and the IPF rat model was constructed,which was divided into the blank control group,IPF model group and Bufei Yiqi Tongluo formula group.The pathological changes of lung tissue in various groups were observed,and the levels of white blood cells (WBC) in bronchoalveolar lavage fluid (BALF) were analyzed. The enzyme-linked immunosorbent assay (ELISA) was used to detect the protein expression levels of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),phosphorylated JNK (p-JNK)/JNK,TGF-β1,tissue inhibitor of metalloproteinases-1 (TIMP-1) and matrix metalloproteinase-9 (MMP-9) in serum,BALF and lung tissue. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression levels of these proteins in serum,BALF and lung tissue.Results The network pharmacology enrichment analysis re-sults revealed that TGF-β1 and JNK signaling pathways were closely related with IPF.Compared with the IPF model group,the lung tissue orderliness in the Bufei Yiqi Tongluo formula group was increased,the consolida-tion range was decreased,the fibrotic tissue proliferation was significantly reduced,the WBC level and expres-sion levels of TNF-α and IL-6 in BALF were significantly decreased.Compared with the blank control group,the mRNA expression levels of TGF-β1,p-JNK/JNK,TIMP-1 and MMP-9 in serum,BALF and lung tissues in the IPF model group were increased with statistical difference (P<0.05).After the intervention on the model rats by the Bufei Yiqi Tongluo formula,except there was no difference in the expression level of MMP-9 pro-tein in serum,TGF-β1 in BALF,TIMP-1 in serum and MMP-9 mRNA in lung tissue,but the other indicators were decreased (P<0.05).Conclusion The therapeutic effect of Bufei Yiqi Tongluo formula in IPF may be related to its regulation on the TGF-β1/JNK signaling pathway.

OBJECTIVE To establish an HPLC method for investigating the stability of mandelonitrile in commonly used solvents and quantitation determination of mandelonitrile in Jian’er Qingjie mixture. METHODS The assay was performed on an Agilent TC-C18(250 mm×4.6 mm, 5 μm) column with a mobile phase consisting of acetonitrile-0.1% phosphoric acid(23∶27) pumped at a flow rate of 1.0 mL·min–1. The column temperature was 30 ℃ and the detection wavelength was set at 207 nm. The stability of the mandelonitrile solution prepared with solvents such as methanol, 95% ethanol, acetonitrile, water, phosphoric acid solution with pH 2.0−6.0, and acetonitrile solution containing 1.0% glacial acetic acid was investigated using the peak reduction rate as the indicator. RESULTS Mandelonitrile was labile in methanol, 95% ethanol or water and relatively stable in acetonitrile. The standard solutions of mandelonitrile prepared with phosphoric acid solutions at pH 2.0−3.5 or acetonitrile containing 1.0% glacial acetic acid were stable in 12 h. The linear range of mandelonitrile was 1.033−294.987 µg·mL–1(r=0.999 9) and the average recovery was 97.4% with RSD of 0.6%(n=9). The content range of mandelonitrile in 16 batches of Jian’er Qingjie mixture produced by 5 manufactures was 3.854−154.578 µg·mL–1. CONCLUSION The established method is simple and accurate. It can be used for the quality control of Jian’er Qingjie mixture. For almond aromatic water and its preparations, the influence of solvent on stability of mandelonitrile should be noticed.

Objective To evaluate function of human neutrophil peptides(HNPs)for the severity and prognosis of patients with acute respiratory distress syndrome(ARDS).Methods From December 2021 to December 2023,80 ARDS patients were enrolled in the study from the intensive care unit(ICU)of Shenzhen Qianhai Shekou Free Trade Zone Hospital.Plasma samples were taken within 24 hours after admission to ICU,HNPs concentration was then measured with enzyme-linked immunesorbent assay.Differences in HNPs concentration and acute physiology and chronic health evaluation-Ⅱ(APACHE Ⅱ)score etc.Clinical index parameters were compared in survival group(n=48)and death group(n=32)based on their 28-day outcomes.The correlation between HNPs concentration and APACHE Ⅱ score were calculated by Spearman correlation analysis.We analyzed the risk factors for 28 days death of ARDS patients by single-factor Logistic regression,then used the significant risk factors to analyze death odds ratio by multivariate Logistic regression analysis.Results HNPs concentration and APACHE Ⅱ score of death group was significantly increased than those of survival group(P<0.001).HNPs concentration has a positive correlation with APACHE Ⅱ score(r=0.344,P<0.001).HNPs and APACHE Ⅱ were independent prognosis factor of 28-day mortality of ARDS patients.Conclusion Plasma HNPs concentration may be early used to evaluate the severity and prognosis of ARDS patients.

Objective To evaluate function of human neutrophil peptides(HNPs)for the severity and prognosis of patients with acute respiratory distress syndrome(ARDS).Methods From December 2021 to December 2023,80 ARDS patients were enrolled in the study from the intensive care unit(ICU)of Shenzhen Qianhai Shekou Free Trade Zone Hospital.Plasma samples were taken within 24 hours after admission to ICU,HNPs concentration was then measured with enzyme-linked immunesorbent assay.Differences in HNPs concentration and acute physiology and chronic health evaluation-Ⅱ(APACHE Ⅱ)score etc.Clinical index parameters were compared in survival group(n=48)and death group(n=32)based on their 28-day outcomes.The correlation between HNPs concentration and APACHE Ⅱ score were calculated by Spearman correlation analysis.We analyzed the risk factors for 28 days death of ARDS patients by single-factor Logistic regression,then used the significant risk factors to analyze death odds ratio by multivariate Logistic regression analysis.Results HNPs concentration and APACHE Ⅱ score of death group was significantly increased than those of survival group(P<0.001).HNPs concentration has a positive correlation with APACHE Ⅱ score(r=0.344,P<0.001).HNPs and APACHE Ⅱ were independent prognosis factor of 28-day mortality of ARDS patients.Conclusion Plasma HNPs concentration may be early used to evaluate the severity and prognosis of ARDS patients.

Objective:To identify the pathogenic characteristics of a suspected gonadal mosaicism Becker muscular dystrophy (BMD) family, and provide provide basis for pregnancy selection of similar families.Methods:A BMD family admitted to Hunan Jiahui Genetics Hospital from June 2012 to September 2019 was systematically reviewed. The medical history and family history of the proband were checked, and multiplex ligation-dependent probe amplification was used to detect the deletion/duplication of 79 exons of the Duchenne muscular dystrophy (DMD) gene in the proband, fetuses, and parents. Moreover, potential variants were verified by combining PCR amplification, short tandom repeat polymorphic linkage analysis, and real-time fluorescence quantitative PCR. High-quality embryos are screened for transplantation after preimplantation genetic testing for monogenic (PGT-M). And amniotic fluid was collected in the second trimester for prenatal diagnostic verification.Results:According to the phenotype analysis of the proband, the initial clinical diagnosis was BMD, and the exon 45-50 deletion in DMD gene was detected. The mutation was not detected in the mother′s peripheral blood, but when she was pregnant again, the prenatal diagnosis showed that the fetus had the same deletion mutation as the proband. Neither of two vitro embryos tested by PGT-M has the deletion mutation, then single embryo transfer was performed nor was pregnancy successful. After confirmation of prenatal diagnosis during pregnancy, a normal baby girl was born by full-term cesarean section.Conclusions:This BMD family was a family with two consecutive BMD homodeletion mutations, and the mutation of the DMD gene was not detected in the peripheral blood of the proband′s mother and two embryonic cells, suggesting that the mother may be a gonad chimeric carrier of this deletion mutation. The combined application of prenatal diagnosis and PGT-M provides a reference approach to effectively avoid the birth of similar children.

Objective:To explore the effect of the enriched environment(EE)on pyroptosis in rats with cerebral ischemia-reperfusion injury(CIRI).Methods:45 male Sprague-Dawley rats were randomly divided into three groups: a sham surgery(Sham)group, a cerebral ischemia-reperfusion(CIR)group and an enriched environment(EE)group, with 15 rats in each group.Except for the Sham group, the right middle cerebral artery occlusion model was established in the other two groups.After surgery, the EE group was fed in EE, and the other two groups were fed in standard environment.All the rats were assessed using the modified neurological severity score(mNSS)before modeling and on the 1st day, 7th day and 14th day following surgery.On the 14th day after surgery, 2, 3, 5-triphenyltetrazolium chloride(TTC)staining was used to evaluate the infarct volume, hematoxylin and eosin(HE)staining was used to examine pathomorphological changes of the hippocampal CA1 region on the ischemic side of the rats in each group, immunohistochemical assay was used to detect the expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)and cysteinyl aspartate specific proteinase-1(caspase-1)proteins in the CA1 region, and ultrastructural changes in neurons in the CA1 region were observed under transmission electron microscopy.Results:Compared with the Sham group, the mNSS scores of the CIR group and the EE group were significantly higher on the 1st day and 7th day after surgery( P<0.05), but there was no significant difference between the CIR and EE groups( P>0.05). On the 14th day after surgery, compared with the CIR group, the EE group showed a decrease in the mNSS score and the cerebral infarct volume( P<0.05), alleviated pathomorphological changes, decreased expression of NLRP3 and caspase-1 proteins( P<0.05), and alleviated pathological changes of pyroptosis in the ultrastructure of neurons. Conclusions:EE can reduce the damage of neurological function, reduce the cerebral infarct volume, and play a protective role for the brain in CIRI rats.The mechanism may be related to the down-regulation of the expression of NLRP3 and caspase-1 proteins related to the classical pyroptosis pathway, leading to the inhibition of pyroptosis.

Objective: To investigate the classification of atrophic glossitis and to study the correlation between the classification and changes of VitB12, folic acid (FOL) and blood cell parameters Methods: A total of 70 patients with atrophic glossitis (AG) were divided into complex type and simple type according to whether they had ulcer or erosion on the tongue mucosa or not. Another 65 healthy subjects during the same period were collected as the control group. The levels of vitamin B12, FOL and blood cell parameters were statistically analyzed using SPSS 25.0 software package. Results: The levels of vitamin B12, red blood cell count (RBC) (3.52 ± 0.69) × 1012·L-1, hemoglobin (HGB)(11.97 ± 1.70) g·dL-1, white blood cell count (WBC) (4.85 ± 1.16) × 109·L-1, neutrophil count (NEUT) (2.76 ± 0.99) × 109·L-1, lymphocyte count (LYMPH) (1.48 ± 0.44) × 109·L-1 in complex type AG group were lower than those in simple type AG group (P<0.05). The levels of mean red blood cell volume (MCV) (104.90 ± 11.13) fL, mean corpuscular hemoglobin (MCH) (34.83 ± 4.56) pg, mean corpuscular hemoglobin concentration (MCHC) (331.09 ± 13.60) g·L-1 were higher than those in the simple type AG group (P<0.05). There was no significant difference in FOL content between these two groups (P>0.05). The levels of VitB12, MCV, MCH, MCHC, WBC, lymph and neut were correlated with the classification of atrophic glossitis (P < 0.05). Conclusion VitB12 deficiency was more apparent in complex AG, especially in large cell anemia, which correlated with the levels of WBC, NEUT, and LYMPH.