BACKGROUND:Extracellular vesicles are secreted into the extracellular milieu by a wide range of cell types,including tumor cells,under different physiological and pathophysiological conditions,where a wide range of biological signals and cell-to-cell signaling exists.Tumor-derived extracellular vesicles may exacerbate cancer progression,survival,invasion,and promote angiogenesis. OBJECTIVE:To review the research progress of extracellular vesicles in the diagnosis and treatment of oral squamous cell carcinoma. METHODS:Literature search was performed by the first author in PubMed,WanFang,CNKI and other databases with the keywords"EVs,oral squamous cell carcinoma,diagnosis and treatment,biopsy,tissue engineering"in Chinese and English.Finally,63 articles were included for analysis. RESULTS AND CONCLUSION:(1)In oral squamous carcinoma saliva biopsies,extracellular vesicles play a crucial role in the progression of oral squamous cell carcinoma by acting as an information transfer tool between tumor cells and the surrounding microenvironment,carrying a wide range of biomolecules including soluble proteins,lipids,DNA,and RNA.These tiny vesicles not only play a key role in tumor growth and spread,but also provide important information about the biological properties of the tumor.(2)Saliva biopsy,as a non-invasive diagnostic method,can open up new possibilities for early diagnosis and targeted treatment of oral squamous cell carcinoma by analyzing the extracellular vesicles therein.(3)It has been found that bioactive molecules,such as microRNAs(miRNAs)and specific proteins,contained within extracellular vesicles can serve as biomarkers for oral squamous carcinoma and improve the accuracy of early diagnosis.Specific proteins in extracellular vesicles such as EHD2,CAVIN1,PF4V1,and CXCL7 show potential as novel predictive biomarkers.(4)In addition,this paper highlights the potential application of extracellular vesicles in the treatment of oral squamous carcinoma.Through engineering modifications,extracellular vesicles can serve as a new generation of nanoscale drug delivery systems to enhance the efficiency and specificity of targeted tumor therapy.(5)Future studies will further explore the effect and mechanism of extracellular vesicles in oral squamous cell carcinoma,which is expected to improve patients'survival and quality of life.

Background and purpose: Long non-coding RNA small nucleolar RNA host gene 5 (lncRNA SNHG5) plays a cancer-promoting role in many cancers, however its effect on colorectal cancer (CRC) and its regulatory mechanism are not clear. This study aimed to explore the mechanism of lncRNA SNHG5/miR-26a-5p/metadherin (MTDH) signal axis promoting metastasis of CRC. Methods: The data of The Cancer Genome Atlas (TCGA) database was analyzed, the abnormal expression of lncRNA in CRC was explored and analyzed the survival. Samples of CRC, paracancerous tissues and complete clinical data of patients who underwent surgical resection from October 2020 to October 2021 were collected. The expression levels of SNHG5 and miR-26a-5p in lncRNA were detected by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR), and the expression level of MTDH was detected by immunohistochemistry. The relationship between the relative expression level of lncRNA SNHG5 in CRC and clinicopathological features and survival time was analyzed. The effects of lncRNA SNHG5 on the proliferation, migration and invasion of CRC cells were detected by cell counting kit-8 (CCK-8), clone formation, scratching assays, transwell test and in vivo xenotransplantation. The relationship between CRC cell metastasis, the expression level of epithelial-mesenchymal transition related molecules and lncRNA SNHG5 expression level by Western blot and immunohistochemical detection were explored. The physical interaction between SNHG5 and miR-26a-5p, MTDH and miR-26a-5p was studied by RNA pull-down test, double luciferase reporter gene detection and RNA co-immunoprecipitation. The functional relationship among the three was verified by CCK-8, EdU and transwell experiments. The effect of SNHG5, miR-26a-5p and MTDH expression on migration and invasion related molecules was analyzed by Western blot. Results: The results of TCGA database analysis showed that lncRNA SNHG5 was significantly upregulated in CRC. The results of RTFQ-PCR and immunohistochemistry showed that the levels of lncRNA SNHG5 and MTDH in CRC tissues were significantly upregulated (P＜0.05), the level of miR-26a-5p was decreased (P＜0.05), and the level of MTDH in samples with high expression of SNHG5 was also increased. The expression of lncRNA SNHG5 in CRC tissues with serosa and extraserosal invasion, distant metastasis, lymph node metastasis and TNM stage Ⅲ was significantly higher compared with subserosal invasion, no distant metastasis and lymph node metastasis and TNM stage Ⅰ-Ⅱ (P＜0.05). The results of survival analysis showed that the high expression of lncRNA SNHG5 was significantly correlated with overall survival rate (P＜0.05). Overexpression of lncRNA SNHG5 could enhance the proliferation, clone formation, migration and invasion of CRC cells, promote the growth and lung metastasis of transplanted tumor, increase the relative expression level of Ki-67 proliferation index and vimentin (P＜0.05), and decrease the relative expression level of E-cadherin (P＜0.05). However, the development of CRC cells was inhibited after inhibition of lncRNA SNHG5 expression. RNA pull-down test, double luciferase reporter gene detection and RNA co-immunoprecipitation confirmed the physical interaction between SNHG5 and miR-26a-5p, MTDH and miR-26a-5p. Upregulation of miR-26a-5p or downregulation of MTDH expression in lncRNA SNHG5 overexpressed cells partially reversed the effects of lncRNA SNHG5 on proliferation, migration, invasion and expression of related molecules in CRC cells. Conclusion: LncRNA SNHG5 is upregulated in CRC tissues and cells, and its high expression is related to tumor progression and poor survival. It can be used as a molecular sponge of miR-26a-5p to regulate the expression of MTDH to promote the proliferation and metastasis of SW620 cells.

Objective:To compare the effect and biotoxicity of tert-butyl acetate (TBA) and ethyl butyrate (EB) on stone dissolution in vitro.Methods:Ten gallstone samples from patients with multiple gallbladder stones were selected and the cholesterol content was analyzed by HPLC. Stone dissolution tests of TBA and EB were performed on cholesterol gallstone in vitro, and the weight of stone at each time point was recorded, meanwhile, methyl tert-butyl ether (MTBE) was used as the control. The inhibitory effects of MTBE, TBA and EB on proliferation of human normal liver cell line LO2 were analyzed by cell proliferation inhibition assay. Flow cytometry was used to analyze the effects of MTBE, TBA and EB on the early and late apoptosis of LO2 cells, and the changes of reactive oxygen species level in LO2 cells were also analyzed.Results:Of the 10 gallbladder gallstones, 6 were cholesterol gallstones and 4 were non-cholesterol gallstones. Stone dissolution experiment showed that the remaining stones of MTBE, TBA and EB groups were (47.83±3.84)%, (58.12±4.53)% and (75.75±4.61)% 30 minutes later. The remaining stones were (18.38±6.47)%, (33.82±6.22)% and (56.38±3.91)% 90 minutes later. MTBE had the best stone dissolution effect in vitro, the stone dissolution effect of TBA was slightly weaker than MTBE, and the stone dissolution effect of EB was relatively weak in all ( P<0.05). The cell proliferation inhibition experiment showed that the cell viability of the control group, MTBE group and TBA group were (100.00±4.46)%, (96.79±4.32)% and (93.72±3.51)%, respectively, and there were no significant differences among the three groups ( P>0.05). However, the cell viability of EB group (87.57±5.29)% was lower than the above three groups, and the differences were statistically significant ( P<0.001). The early apoptosis and late apoptosis of the control group were (1.67±0.15)% and (1.27±0.06)%, respectively. EB induced early apoptosis (15.90±0.53)% ( P<0.001) and late apoptosis (5.13±0.76)% ( P<0.05). However, MTBE and TBA had no significant effect on cell apoptosis ( P>0.05). Compared with control group, MTBE, TBA and EB all significantly inhibited the level of reactive oxygen species ( P<0.05), and the inhibitory effect of EB was the most obvious. Conclusions:TBA has good stone dissolution effect and biosafety for gallbladder cholesterol stones in vitro, while EB has relatively poor performance. TBA is a potential drug for gallstone dissolution.

Oral squamous cell carcinoma(OSCC),a common malignancy of the head and neck,ranks sixth worldwide in terms of cancers with the most negative impact,owing to tumor relapse rates,cervical lymphnode metastasis,and the lack of an efficacious systemic therapy.Its prognosis is poor,and its mortality rate is high.Octamer-binding transcription factor 4(OCT4)is a member of the Pit-Oct-Unc(POU)family and is a key reprogramming factor that produces a marked effect in preserving the pluripotency and self-renewal state of embryonic stem cells(ESCs).According to recent studies,OCT4 participates in retaining the survival of OSCC cancer stem cells(CSCs),which has far-reaching implications for the occurrence,recurrence,metastasis,and prognosis of oral carcinogenesis.Therefore,we summarize the structure,subtypes,and function of OCT4 as well as its role in the occurrence,progression,and prognosis of OSCC.

Background: During hot environment work tasks with whole-body enclosed anti-bioaerosol suit, the combined effect of heavy sweating and exhaled hot humid air may cause the N95 medical respirator to saturate with water/sweat (i.e., water-blocking). Methods: 32 young male subjects with different body mass indexes (BMI) in whole-body protection (N95 medical respirator + one-piece protective suit + head covering + protective face screen + gloves + shoe covers) were asked to simulate waste collecting from each isolated room in a seven-story building at 27-28°C, and the weight, inhalation resistance (Rf), and aerosol penetration of the respirator before worn and after water-blocking were analyzed. Results: All subjects reported water-blocking asphyxia of the N95 respirators within 36-67 min of the task. When water-blocking occurred, the Rf and 10-200 nm total aerosol penetration (Pt) of the respirators reached up to 1270-1810 Pa and 17.3-23.3%, respectively, which were 10 and 8 times of that before wearing. The most penetration particle size of the respirators increased from 49-65 nm before worn to 115-154 nm under water-blocking condition, and the corresponding maximum size-dependent aerosol penetration increased from 2.5-3.5% to 20-27%. With the increase of BMI, the water-blocking occurrence time firstly increased then reduced, while the Rf, Pt, and absorbed water all increased significantly. Conclusions This study reveals respirator water-blocking and its serious negative impacts on respiratory protection. When performing moderate-to-high-load tasks with whole-body protection in a hot environment, it is recommended that respirator be replaced with a new one at least every hour to avoid water-blocking asphyxia.

Background: The mouse model of high-fat diet-induced cholesterol gallstone is widely used in researches of pathogenesis, prevention and treatment of gallstones. Aims: To investigate the characteristics and hepatic transcriptomics of the mouse model of high-fat diet-induced cholesterol gallstone. Methods: Twenty male C57BL/6J mice were randomly allocated into chow diet (control) group and lithogenic diet (LD) group. After 8 weeks, the occurrence of gallstone was observed; the serum lipids and gallbladder bile lipids were detected; and the differentially expressed hepatic genes between the two groups were identified with Illumina NovaSeq sequencing systems. The enrichment analysis was mapped in GO and KEGG pathway databases. Real-time PCR was used to detect the expressions of genes related to bile acid synthesis in the liver. Results: The cholesterol gallstone formation rate was 100% in LD group, whereas no gallstone was observed in control group. Hepatomegaly and steatosis were obvious in mice of LD group. The serum levels of total cholesterol and triacylglycerol, as well as the cholesterol content and cholesterol saturation index of the gallbladder bile in LD group were significantly higher than those in control group (P<0.05). A total of 1 330 differentially expressed genes were identified by high-throughput sequencing and bioinformatics analysis. KEGG analysis showed that the differentially expressed genes were mainly enriched in lipid metabolism, bile secretion, and insulin secretion pathways. GO analysis showed that fatty acid metabolic process-related pathways were significantly enriched. Both hepatic transcriptomics analysis and real-time PCR showed that the expression levels of genes related to bile acid synthesis, including CYP7A1, CYP8B1 and CYP7B1 decreased significantly in the liver of LD group as compared with those of control group (P<0.05). Conclusions: The metabolism of cholesterol, bile acids and fatty acids is significantly disordered in mice with cholesterol gallstone. Transcriptomics analysis can screen out the differentially expressed genes that play roles in the formation of cholesterol gallstone, so as to provide references for studies focusing on these topics.

Objective: To investigate the clinicopathologic features, immunophenotype, pathological diagnosis and treatment of malignant mixed tumor (MMT). Methods: Clinical and pathological features including immunohistochemical phenotypes were analyzed in a case of MMT accompanied with eccrine porocarcinoma (EP) involving both hands, diagnosed definitely in January 2018 along with review of relevant literature. Results: A 64-year-old man presented with multiple rash on both hands for 4 years. Three lesions of 0.5 to 2.2 cm were removed for pathological evaluation. The pathological changes on little finger of left and right hands were MMT with EP, whereas that removed from the right ring finger was EP. MMT showed infiltrative growth with vascular wall invasion and consisted of epithelial (glandular or tube differentiation) and mesenchymal components (mucinous and/or cartilage stroma). The endothelial cells showed moderate to severe cytological atypia, nuclear pleomorphism and increased mitotic activity. The glandular component had histological characteristics of syringocarcinoma with moderately atypical chondrocytes but without myoepithelium. EP was composed of basal cells with visible vacuoles in cytoplasm and the presence of tubular and squamous differentiation, along with obvious atypia. Immunohistochemically cavosurface epithelium of glandular differentiation of MMT showed positivity for CK7, EMA and CD117. Myoepithelium showed S-100, CK5/6 and p63 positivity and stromal cells were positive for S-100. Differential diagnoses included metaplastic carcinoma, malignant myoepithelioma and atypical mixed tumor of skin. Conclusions MMT with EP is extremely rare.The diagnosis of MMT depends on the morphologic features. Immunohistochemical staining is helpful for differential diagnosis. Surgical excision with safety margins is the treatment of choice. Complementary radiotherapy and/or chemotherapy is still controversial. The clinical course of MMT is deemed unpredictable and long-term follow-up is necessary.

Objectives To investigate the occipital cortex metabolite alterations in repetitive and severe neonatal hypoglycemia rats treated with sodium pyruvate and to reveal the protective role of sodium pyruvate using high resolution 1H nuclear magnetic resonance spectroscopy.Methods Thirty-six 2-dayold Sprague-Dawley rats were randomly divided into hypoglycemia group and pyruvate group with 18 rats in each group.Rats in both groups received intraperitoneal injections of insulin (40 U/kg body weight) at 2,4 and 6 days of age to induce severe hypoglycemia (blood glucose value ≤ 1.4 mmol/L).In the hypoglycemia group,2.5 hours after insulin injection,intraperitoneal injection of 50％ glucose (2 ml/kg) was administered to terminate hypoglycemia,while in the pyruvate group,50％ glucose (2 ml/kg) and sodium pyruvate solution 2.5 ml/kg (500 mg/kg) were injected.Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay was used to observe the status of injured neurons in six neonatal rats,and metabolite changes in occipital cortex of the other 12 rats were detected by 1H nuclear magnetic resonance spectroscopy.The difference between the two groups was compared by independent-samples t test.Results Neonatal rats of both groups reached severe hypoglycemia level 2.5 hours after insulin injection.Compared with hypoglycemia group,pyruvate group had fewer injured neurons (45±5 vs 113 ± 12,t=0.782,P=0.013) and lower injured index in the occipital cortex (0.15 ± 0.03 vs 0.36 ± 0.06,t=l.143,P=0.020).Pyruvate group showed significant decreases in the concentration of taurine [(13.31 ± 2.06) vs (18.44 ± 3.86) mol/kg,t=8.231],glutamine[(1.50 ± 0.24) vs (2.02 ± 0.40) mol/kg,t=3.137],glutamate[(7.04 ± 0.95) vs (9.40 ± 1.73) mol/kg,t=6.449],aspartate[(1.51 ± 0.28) vs (2.15 ± 0.58) mol/kg,t=2.561] and creatine [(6.37±0.99) vs (8.46± 1.77) mol/kg,t =4.226] in the occipital cortex (all P'＜0.017).Conclusions Simultaneous use of glucose and sodium pyruvate to terminate hypoglycemia in repetitive and severe neonatal hypoglycemia rats can effectively alleviate severe hypoglycemia-induced occipital lobe damage via regulating excitatory amino acid neurotransmitters,energy metabolism and other metabolic pathways.

Objective To investigate certain risk factors for and their impact on abnormal liver function in middle-aged and elderly adults.Methods A case-control study was constructed based on the SAGE cohort of 8642 registered residents aged 50 years or over in Shanghai.Of them,137 individuals with abnormal liver function,defined as aspartate transaminase (AST)＞ 40 U/L or alanine aminotransferase (ALT)＞ 40 U/L,were randomly selected as the observation group,while 411 healthy controls were 3 ∶ 1 matched with the cases in the observation group by gender and age (1 year).Face-to-face administered questionnaires and physical examinations were conducted and serum samples were tested for ALT,AST,glucose (GLU),total cholesterol (TC),triglycreide (TG),hepatitis B surface antigen (HbsAg) and anti-hepatitis C virus antibody (anti-HCV Ab).Chi square test and rank sum test were used for single factor analysis,and logistic regression analysis was used for multiple factors.Results The prevalence of HBsAg positive patients was 12.4 ％ (68/548) Univariate analysis showed that hepatitis virus infection and body mass index (BMI) were associated with abnormal liver function (both P＜0.000).Multivariate logistic regression analysis showed that hepatitis virus infection (OR=1.85,95％ CI:1.04 3.29,P-0.036) and obesity (OR=3.60,95％CI:1.92-6.73,P＜0.001) increased the risk of abnormal liver function,whereas chronic medication (OR=0.51,95％ CI:0.32-0.80,P =0.004) decreased the risk of abnormal liver function.Conclusions Among the study population,hepatitis virus infection and obesity are risk factors for abnormal liver function in middle-aged and elderly people.After adjustment for potential confounders,chronic medication is negatively correlated with abnormal liver function and may be a protective factor for liver function.

ObjectiveTo investigate the clinical promoting effect of crossing nape electroacupuncture on the recovery of swallowing function and recovery from pulmonary infection in post-cerebral infarction patients with tracheotomy.MethodSixty post-cerebral infarction patients with cough reflex disorder and swallowing dysfunction associated with pulmonary infection receiving tracheotomy and tracheal intubation were subjects. They were allocated, using a random number table, to three groups, 20 cases each. In each group, the patients were enrolled in order of visits. The three groups were given the same basic treatment for fighting inflammation, resolving phlegm and improving blood supply. The crossing nape electroacupuncture group received bilateral points Fengchi (GB20), Yifeng (TE17), Dicang (ST4)-to-Jiache (ST6) and Lianquan (CV23) acupuncture with electrodes connected by left-right crossing. The acupuncture group received bilateral points Fengchi, Yifeng, Dicang-to-Jiache and Lianquan acupuncture without electrodes connected. The control group received basic treatment with Western drugs without acupuncture therapy. Observations were carried out using the Kubota’s water drinking test, the Toshima Ichiro Swallowing Assessment and the Clinical Pulmonary Infection Score. The clinical therapeutic effects were evaluated in the three groups.ResultThe therapeutic effects evaluated using the Kubota’s water drinking test and the Toshima Ichiro Swallowing Assessment were better in the crossing nape electroacupuncture group than in the acupuncture group and better in the acupuncture group than in the control group (P<0.05). The score of the Clinical Pulmonary Infection Score decreased in all the three groups. The promoting effect on recovery from pulmonary infection was marked in the crossing nape electroacupuncture group (P<0.01).ConclusionCrossing nape electroacupuncture has a marked improving effect on dysphagia in post-cerebral infarction patients with tracheotomy and tracheal intubation. It can promote recovery from pulmonary infection in post-cerebral infarction patients with cough reflex disorder receiving tracheotomy and tracheal intubation.