Objective: To explore the mechanism of action of hepatic leukemia factor (HLF) in diabetes mellitus and to construct a conditional animal model of mice with islet β ⁃cell⁃specific HLF gene knockout. Methods: At the cellular level , the effects of HLF inhibition or overexpression on the proliferation of MIN6 cells was verified by the CCK⁃8 assay. The effects of HLF inhibition or overexpression were detected at the mRNA level and protein level by Cre + / - mice (C57BL/6J) to obtain offspring mice. The genotypes of the mice were identified by the PCR method.The differences in the expression levels of the HLF gene at the mRNA and protein levels in islet β ⁃cell knockout mice (HLFflox/flox Cre + / - ) and control mice (HLFflox/flox ) were detected by RT⁃qPCR technology and Western blot technology to verify the knockout effect. At the same time , the islet tissues of the mice in two groups were taken to make paraffin sections and analyzed by hematoxylin⁃eosin (HE) staining. Results: HLF gene inhibition or overex⁃pression had no significant effect on the proliferation of MIN6 cells. When the HLF gene was inhibited in MIN6 cells , the mRNA expression level decreased by 74% compared with the control group , and the protein expression level decreased by 60% compared with the control group. After overexpressing the HLF gene , the mRNA expres⁃sion level was 2. 13 times compared with that of the control group , and the protein expression level was 1. 8 times compared with that of the control group. The mRNA expression level of the HLF gene in the knockout mice de⁃creased by 89% compared with the control group , and the protein expression level decreased by 65% compared with the control group. The results of HE staining showed that there was no significant difference in the cell mor⁃phology in the islet tissues between the knockout mice and the control mice. Inhibiting HLF increased the glycogen content in MIN6 cells by approximately 20% . Conclusion The HLF gene knockout mice are successfully con⁃structed , providing an animal model for studying the role of HLF in the pathogenesis of diabetes mellitus.

Objective To explore the effect of dexmedetomidine on the proliferation,invasion and cell cycle of gallbladder cancer cells by regulating the C-C chemokine ligand 2(CCL2)-C-C chemokine receptor 2(CCR2)pathway.Methods GBC-SD cells were devided into the control group,the low concentration dexmedetomidine group(2 μmol/L),the high concentration dexmedetomidine group(4 μmol/L)and the high concentration dexmedetomidine+CCL2 group(4 μmol/L dexmedetomidine and 10 μg/L CCL2 protein).The clone formation experiment and Edu experiment were performed to measure cell proliferation.Transwell experiment was performed to measure cell invasion and migration.Flow cytometry was performed to measure cell cycle and apoptosis.Western blot assay was used to measure the proliferating cell nuclear antigen(PCNA),Cyclin D1,matrix metalloproteinase(MMP)-2,MMP-9,Bcl-2 associated X protein(Bax),cysteine-dependent aspartate-specific protease-3(Caspase-3),CCL2 and CCR2 proteins.The nude mouse transplant tumor experiment was used to determine the growth of gallbladder cancer transplant tumors.Results After treatment with low and high concentrations of dexmedetomidine,the number of cell clone formed,the positive rate of Edu,the numbers of invasions and migrations,the expression levels of PCNA,CyclinD1,MMP-2,MMP-9,CCL2 and CCR2 proteins,the proportions of G2/M and S phase cells decreased,the proportion of G0/G1 phase cells,apoptosis rate and expression levels of Bax and Caspase-3 proteins increased,and the effect of high-concentration dexmedetomidine was more significant(P＜0.05).The inhibitory effects of dexmedetomidine on the proliferation,invasion,migration and cell cycle of gallbladder cancer cells,as well as its promoting effect on cell apoptosis could be reversed by CCL2 protein(P＜0.05).In vivo experiments showed that dexmedetomidine could reduce tumor mass,tumor volume of nude mice and expression levels of CCL2 and CCR2 proteins(P＜0.05).Conclusion Dexmedetomidine inhibits the proliferation and invasion of gallbladder cancer cells,and blocks the cell cycle in the G0/G1 phase by suppressing the CCL2-CCR2 pathway.

Objective:To evaluate the clinical effect of transforaminal endoscopic spine system (TESSYS) technique in treating single-level lumbar spinal stenosis (LSS) based on computed tomography (CT) imaging and traumatic stress indicators. Methods:A total of 112 patients with single-level LSS admitted to The Third Hospital of Qinhuangdao from June 2019 to June 2021 were selected,and they were divided into observation group and control group by random number table method,with 56 cases in each group. The observation group was treated with TESSYS technique,and the control group was treated with "broad easy immediate surgery (BEIS)" technique of intervertebral foramen scope. The indicators of CT imaging of spinal canal area and protrusion ratio,as well as traumatic stress indicators included lactate dehydrogenase (LDH),myoglobin (MYO) and cortisol (COR),of two groups were compared before and 1 week after surgery. Oswestry Disability Index (ODI),Visual Analogue Scale (VAS) score and Japanese Orthopaedic Association (JOA) Scores were compared between the two groups. Excellent rates of treatment and postoperative complications of two groups were compared at the 6 th month after surgery. Results:The average hospital stay and operation time of observation group were respectively (4.29±1.08)d and (93.53±22.01)min,which were significantly shorter than (6.61±1.72)d and (112.29±26.68)min of control group,and the intraoperative blood loss of observation group was (30.15±8.26) ml,which was shorter than that (41.35±11.58) ml of control group,and the differences of them between two groups (t=8.548,4.059,5.892,P<0.05),respectively. At the 1st week after surgery,the ODI and VAS scores of observation group were lower than those of control group,and the JOA score was higher than that of control group,and the differences of them between two groups were significant (t=9.443,t=8.969,6.084,P<0.05),respectively. At the 3rd month after surgery,the ODI and VAS scores of observation group were respectively lower than those of control group,and JOA score of observation group was higher than that of control group,and the differences of them were significant (t=9.706,3.753,5.894,P<0.05),respectively. The rate of excellent and good treatment of observation group was 94.64％,which was higher than that (80.36％) of control group,and the differences of them between the two groups were statistically significant (t=5.225,P<0.05). At the 1st week after surgery,the vertebral canal area of observation group was larger than that of control group,and the ratio of protrusion invasion was less than that of control group,and the differences of them between the two groups were statistically significant (t=3.404,5.578,P<0.05),respectively. At the 1st d after surgery,the serum LDH,MYO and COR levels of observation group were lower than those of control group,and the differences of them between two groups were statistically significant (t=7.570,9.687,5.242,P<0.05),respectively. At the 3rd d after surgery,the serum LDH,MYO and COR levels of observation group were respectively than those of control group,and the differences of them between two groups were significant (t=6.856,9.729,2.744,P<0.05). There was no significant difference in complication rate between the two groups (P>0.05). Conclusion:Compared with BEIS technique,TESSYS technique has the advantages of less trauma,faster postoperative recovery and higher excellent and good rate in treating single-segment LSS,which can alleviate stress reaction of early postoperative trauma. Its comprehensively clinical efficacy is ideal.

Objective:To evaluate the clinical effect of transforaminal endoscopic spine system (TESSYS) technique in treating single-level lumbar spinal stenosis (LSS) based on computed tomography (CT) imaging and traumatic stress indicators. Methods:A total of 112 patients with single-level LSS admitted to The Third Hospital of Qinhuangdao from June 2019 to June 2021 were selected,and they were divided into observation group and control group by random number table method,with 56 cases in each group. The observation group was treated with TESSYS technique,and the control group was treated with "broad easy immediate surgery (BEIS)" technique of intervertebral foramen scope. The indicators of CT imaging of spinal canal area and protrusion ratio,as well as traumatic stress indicators included lactate dehydrogenase (LDH),myoglobin (MYO) and cortisol (COR),of two groups were compared before and 1 week after surgery. Oswestry Disability Index (ODI),Visual Analogue Scale (VAS) score and Japanese Orthopaedic Association (JOA) Scores were compared between the two groups. Excellent rates of treatment and postoperative complications of two groups were compared at the 6 th month after surgery. Results:The average hospital stay and operation time of observation group were respectively (4.29±1.08)d and (93.53±22.01)min,which were significantly shorter than (6.61±1.72)d and (112.29±26.68)min of control group,and the intraoperative blood loss of observation group was (30.15±8.26) ml,which was shorter than that (41.35±11.58) ml of control group,and the differences of them between two groups (t=8.548,4.059,5.892,P<0.05),respectively. At the 1st week after surgery,the ODI and VAS scores of observation group were lower than those of control group,and the JOA score was higher than that of control group,and the differences of them between two groups were significant (t=9.443,t=8.969,6.084,P<0.05),respectively. At the 3rd month after surgery,the ODI and VAS scores of observation group were respectively lower than those of control group,and JOA score of observation group was higher than that of control group,and the differences of them were significant (t=9.706,3.753,5.894,P<0.05),respectively. The rate of excellent and good treatment of observation group was 94.64％,which was higher than that (80.36％) of control group,and the differences of them between the two groups were statistically significant (t=5.225,P<0.05). At the 1st week after surgery,the vertebral canal area of observation group was larger than that of control group,and the ratio of protrusion invasion was less than that of control group,and the differences of them between the two groups were statistically significant (t=3.404,5.578,P<0.05),respectively. At the 1st d after surgery,the serum LDH,MYO and COR levels of observation group were lower than those of control group,and the differences of them between two groups were statistically significant (t=7.570,9.687,5.242,P<0.05),respectively. At the 3rd d after surgery,the serum LDH,MYO and COR levels of observation group were respectively than those of control group,and the differences of them between two groups were significant (t=6.856,9.729,2.744,P<0.05). There was no significant difference in complication rate between the two groups (P>0.05). Conclusion:Compared with BEIS technique,TESSYS technique has the advantages of less trauma,faster postoperative recovery and higher excellent and good rate in treating single-segment LSS,which can alleviate stress reaction of early postoperative trauma. Its comprehensively clinical efficacy is ideal.

Objective To explore the effect of dexmedetomidine on the proliferation,invasion and cell cycle of gallbladder cancer cells by regulating the C-C chemokine ligand 2(CCL2)-C-C chemokine receptor 2(CCR2)pathway.Methods GBC-SD cells were devided into the control group,the low concentration dexmedetomidine group(2 μmol/L),the high concentration dexmedetomidine group(4 μmol/L)and the high concentration dexmedetomidine+CCL2 group(4 μmol/L dexmedetomidine and 10 μg/L CCL2 protein).The clone formation experiment and Edu experiment were performed to measure cell proliferation.Transwell experiment was performed to measure cell invasion and migration.Flow cytometry was performed to measure cell cycle and apoptosis.Western blot assay was used to measure the proliferating cell nuclear antigen(PCNA),Cyclin D1,matrix metalloproteinase(MMP)-2,MMP-9,Bcl-2 associated X protein(Bax),cysteine-dependent aspartate-specific protease-3(Caspase-3),CCL2 and CCR2 proteins.The nude mouse transplant tumor experiment was used to determine the growth of gallbladder cancer transplant tumors.Results After treatment with low and high concentrations of dexmedetomidine,the number of cell clone formed,the positive rate of Edu,the numbers of invasions and migrations,the expression levels of PCNA,CyclinD1,MMP-2,MMP-9,CCL2 and CCR2 proteins,the proportions of G2/M and S phase cells decreased,the proportion of G0/G1 phase cells,apoptosis rate and expression levels of Bax and Caspase-3 proteins increased,and the effect of high-concentration dexmedetomidine was more significant(P＜0.05).The inhibitory effects of dexmedetomidine on the proliferation,invasion,migration and cell cycle of gallbladder cancer cells,as well as its promoting effect on cell apoptosis could be reversed by CCL2 protein(P＜0.05).In vivo experiments showed that dexmedetomidine could reduce tumor mass,tumor volume of nude mice and expression levels of CCL2 and CCR2 proteins(P＜0.05).Conclusion Dexmedetomidine inhibits the proliferation and invasion of gallbladder cancer cells,and blocks the cell cycle in the G0/G1 phase by suppressing the CCL2-CCR2 pathway.

Objective To investigate the effects of polybutylene terephthalate(PBT)grafted with various monomers,such as acrylic acid(AA),acrylamide(AM),sodium styrene sulfonate(SSS),AA+AM and AA+SSS on platelet adhesion and function.Methods The AA,AM,SSS,AA+AM,and AA+SSS were grafted onto the surface of PBT by γ-ray irradia-tion,and the grafted PBT was characterized by fourier transform infrared spectrometry(FTIR)and wetting time.Platelet ac-tivation and aggregation of different monomers grafted with PBT were observed by scanning electron microscopy(SEM).Platelet concentration,maximum aggregation ability,positive expression rate of CD62p and hypotonic shock rate(HSR)of PBT grafted with different monomers were tested to study their effect on platelet adhesion and function.Results The char-acteristic absorption peaks of AA,AM and SSS appeared in FTIR,and the hydrophilicity of the grafted material was im-proved obviously,indicating that the monomer was grafted successfully.The results of SEM showed that the degree of platelet activation and aggregation caused by the original PBT was significantly higher than that of modified ones.Compared with the original platelets,the platelet concentration of PBT was(533.00±4.58 vs 672.00±3.61)×109/L,the maximum platelet ag-gregation rate was(48.80±0.96 vs 58.60±1.37)%,the positive expression rate of CD62p was(45.35±0.58 vs 39.90±0.52)%,and the platelet HSR was(48.74±0.46 vs 51.86±0.93)%(P<0.05).Compared with the original PBT,the platelet loss and platelet function damage caused by PBT grafted with different monomers decreased significantly(P<0.05).PBT-(AA+AM)had the least comprehensive effect on platelets[platelet concentration(637.00±2.65)×109/L,maximum plate-let aggregation rate(62.45±0.61)%,positive expression rate of CD62p(37.39±0.42)%,platelet HSR(53.51±0.58)%].Conclusion The comprehensive effect of PBT grafted with AA+AM on platelet adhesion and function is significantly lower than that of PBT grafted with other monomers,so it is anticipated to apply filtering and removing leukocytes in platelet prep-arations.

N1-methyladenosine(m1A)RNA methylation is critical for regulating mRNA translation;however,its role in the development,progression,and immunotherapy response of head and neck squamous cell carcinoma(HNSCC)remains largely unknown.Using Tgfbr1 and Pten conditional knockout(2cKO)mice,we found the neoplastic transformation of oral mucosa was accompanied by increased m1A modification levels.Analysis of m1A-associated genes identified TRMT61A as a key m1A writer linked to cancer progression and poor prognosis.Mechanistically,TRMT61A-mediated tRNA-m1A modification promotes MYC protein synthesis,upregulating programmed death-ligand 1(PD-L1)expression.Moreover,m1A modification levels were also elevated in tumors treated with oncolytic herpes simplex virus(oHSV),contributing to reactive PD-L1 upregulation.Therapeutic m1A inhibition sustained oHSV-induced antitumor immunity and reduced tumor growth,representing a promising strategy to alleviate resistance.These findings indicate that m1A inhibition can prevent immune escape after oHSV therapy by reducing PD-L1 expression,providing a mutually reinforcing combination immunotherapy approach.

The objectification of facial diagnosis has been developed through recent years and has become a multidisciplinary research topic.However,many studies are still limited to the adjustment of algorithms and the design of data collection environment and equipments,few studies focus on facial diagnosis zoning.The purpose of this paper is to discuss the problems in the current research literature on machine learning-based intelligent TCM facial diagnosis zoning to build a foundation for subsequent related research.The study uses bibliometric methods and text analysis to clarify and analyze the current intelligent TCM facial diagnosis zoning methods,which mainly include facial feature point-based,facial feature block-based and complete face-based method;then by analyzing the influencing factors of facial diagnosis zoning research and summarizing the common machine learning algorithms,the advantages and disadvantages of different machine learning algorithms and the corresponding common facial diagnosis zoning methods are obtained;Finally,we discuss three aspects of the current phase of facial diagnosis zoning research:dataset construction,advantages of deep learning,and embodiment of facial diagnosis theory.

Objective To investigate the effect of sufentanil(ST)on lipopolysaccharide-induced acute lung injury(ALI)in rats by regulating the cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)/stimulator of interferon gene(STING)signaling pathway.Methods Male SD rats were induced by lipopolysaccharide to construct an ALI model.The successfully modeled rats were randomly separated into ALI group,L-ST group,H-ST group,and H-ST+DMXAA group,with 6 rats in each group.Among them,the L-ST group and H-ST group were intraperitoneally injected with 2.5 μg/ml and 5 μg/ml ST after successful modeling immediately.The H-ST+DMXAA group was given 25 mg/kg DMXAA by gavage on the basis of intraperitoneal injection of 5 μg/ml ST.Six healthy and normal temperature rats were randomly selected as the control group,and an equal amount of physiological saline was injected intraperitoneally.The reagent kit was used to detect the SOD enzyme activity and MDA content in serum;ELISA was used to detect the levels of inflammatory factors such as interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),and IL-6 in bronchoalveolar lavage fluid(BALF);cell staining was used to detect the total number of cells and neutrophils in BALF;the wet to dry mass ratio of lung tissue was calculated;HE staining was used to observe pathological changes in lung tissue,while Western blot was applied to detect the expression of cGAS and STING proteins in lung tissue.Results After L-ST and H-ST treatment,the SOD enzyme activity and IL-10 level increased sequentially,the MDA,TNF-α,IL-6,W/D value,total number of cells and neutrophils,cGAS,and STING protein expression decreased sequentially(P<0.05),while lung tissue injury was effectively alleviated.DMXAA reversed the influence of H-ST on the above indexes(P<0.05).Conclusion ST may inhibit the cGAS-STING pathway,reduce inflammation and oxidative stress responses,and thereby alleviates lipopolysaccharide induced ALI in rats.