BACKGROUND:Solute carrier family 1 member 5(SLC1A5)plays a potential role in a variety of diseases,but the exact mechanism of action is unclear.The construction of stable SLC1A5 overexpression and knockdown cell models can provide a powerful experimental tool for in-depth study of the exact role and mechanism of SLC1A5 in diseases and the discovery of potential therapeutic targets. OBJECTIVE:To construct lentiviral vectors for overexpression and knockdown of mouse SLC1A5 and establish stable transfected RAW264.7 cell lines,so as to provide an experimental foundation for further investigation of the role of SLC1A5 in inflammation. METHODS:Primers were designed and synthesized based on the SLC1A5 gene sequence,and the gene segment was amplified using polymerase chain reaction.Subsequently,the target gene segment was directionally inserted into the GV492 vector plasmid,which had been digested with AgeI/NheI enzymes,to construct recombinant lentiviral plasmids.Positive clones were further selected,and their sequences were confirmed.The pHelper1.0 plasmid vector and pHelper2.0 plasmid vector,along with the target plasmid vector,was co-cultured with 293T cells for transfection,resulting in the production and titration of lentiviral stocks.Furthermore,RAW264.7 cells were cultured in vitro,and the working concentration of puromycin was determined.Lentiviruses were separately co-cultured with RAW264.7 cells,and transfection efficiency was determined by measuring fluorescence intensity.Stable transfected cells were selected using puromycin,and real-time fluorescence quantitative PCR and western blot assay were employed to assess the gene and protein expression levels of SLC1A5 in stably transfected cell lines. RESULTS AND CONCLUSION:(1)Sequencing results indicated a perfect match between the sequencing and target sequences,confirming the successful construction of recombinant lentiviral vectors.(2)The titer for the overexpression SLC1A5 lentivirus was 1×109 TU/mL,while the titer for the knockdown SLC1A5 lentivirus was 3×109 TU/mL.(3)The working concentration of puromycin for RAW264.7 cells was determined to be 3 μg/mL.(4)The optimal conditions for transfecting RAW264.7 cells with overexpression/knockdown expression of SLC1A5 lentivirus involved the use of HiTransG P transfection enhancer with a multiplicity of infection value of 50.(5)A significant upregulation of the gene and protein expression levels of SLC1A5 was detected in cell lines stably overexpressing SLC1A5,while gene and protein expression levels of SLC1A5 were significantly decreased in the knockdown stable cell lines.These findings indicate that lentiviral vectors for mouse SLC1A5 overexpression and knockdown have been successfully constructed and a stably transfected RAW264.7 cell line has been obtained.

To investigate the effects of Wogonin (WO) on learning and memory impairment, Aβ1-42 was injected intracerebroventricularly to induced a mouse learning and memory impairment model, and D-galactose was injected intraperitoneally to induced a mouse acute aging model. Mice were administered WO (75, 150, or 300 mg/kg) by oral gavage for 28 consecutive days. Cognitive function was assessed using the Morris water maze (MWM), novel object recognition (NOR), and open field tests (OFT). In the Aβ1-42 model, WO treatment (150 and 300 mg/kg) significantly improved the recognition index in the NOR test, while the 150 mg/kg group showed increased target quadrant preference in the MWM test. No changes in the total distance traveled in OFT. In the D-galactose aging model, the 150 mg/kg WO group exhibited increased platform crossings in the MWM test, and all WO doses (75, 150, and 300 mg/kg) enhanced target quadrant preference, with no alterations in spontaneous movement. Western blot analysis revealed that WO significantly attenuated hippocampal apoptosis in both models. These findings suggest that WO ameliorates learning and memory impairment associated with Alzheimer’s disease and aging.

Purpose To investigate the clinicopathological features,molecular genetics and prognosis of high grade B cell lymphoma with concurrent MYC rearrangement and 11q aberra-tions(HGBCL-MYC-11q).MethodsThree cases of HGBCL-MYC-11q were reviewed and analyzed using hematoxylin-eosin staining,immunohistochemistry,EBER in situ hybridization and fluorescence in situ hybridization.Clinical data were collected with follow-up.Results All three patients were male,age was 10,61,and 74 years,respectively.All patients had Ann Arbor stage Ⅳ disease.All three cases were biopsies occurring in the nasopharynx,upper pharynx and ileocecus,respectively.Three cases were morphologically similar to diffuse infiltrative growth of tumor cells,moderate or moderately large cells,round to slightly irregular nuclei and easily visible mitotic figures.Focal necrosis was noted in one case.One case exhibited the distinct"starry sky"pattern.All cases expressed CD20,BCL6 and MUM1 and high Ki67 index,two cases expressed CD10 and two cases ex-pressed BCL2.CD3,CD30 and TDT were all negative.EBER in situ hybridization was all negative.FISH analyses using C-MYC break-apart probes were all positive and all cases had 11q aberrations.One case only had the 11q23.3 amplification;and one case only had the 11q24.3 loss.After a follow-up for 1-18 months,one patient died and two patients survived with disease.ConclusionHGBCL-MYC-11q is rare,morphologically similar to BL/HGBCL,with MYC rearrangement and 11q abnormali-ties.We should enhance awareness of the disease and improve more accurate diagnosis and differential diagnosis of the disease.

Objective To investigate the incidence and influencing factors of aggressive behavior in adolescent inpatients with mental disorders.Methods A total of 372 patients with mental disorders admitted to the Department of Child and Adolescent Psychiatry of the Second Affiliated Hospital of Xinxiang Medical University from January to December 2022 were selected as research subjects.The occurrence of aggressive behavior and its influencing factors were analyzed.Results Among the 372 patients,38(10.2％)cases had aggressive behavior during hospitalization.Univariate and multivariate logistic regression results showed that the history of aggressive behavior,only child,and high irritation factor scores were the risk factors of aggressive behavior in hospitalized adolescents with mental disorders(P＜0.05).Mild,outgoing,insecure and isolated personalities before the onset were protective factors of aggressive behavior in hospitalized adolescents with mental disorders(P＜0.05).Conclusion Adolescent inpatients with mental disorders are likely to have aggressive behavior.Those who have aggressive behavior before,are the only child in their family and get a high score of irritant factors need more attention.Corresponding nursing interventions should be taken in time through nursing assessment to reduce the occurrence of aggressive behavior.

Purpose To detect the expression of PP1A and GSDME and the abundance of CD8+T lymphocytes in colorectal cancer,and to explore the correlation and clinical significance of PP1A and GSDME mediated pyroptosis.Methods GEPIA da-tabase was applied to analyze the mRNA expression of PP1A and GSDME in colorectal cancer and normal tissues.Western blot assay was used to detect the expression of PP1A in colorectal cancer and the corresponding normal mucosa.Immunohisto-chemistry was applied to detect the expression of PP1A and GS-DME and CD8+T lymphocytes abundance in 107 colorectal car-cinomas and normal mucosa adjacent to the carcinomas.Spearman rank correlation was used to analyze the correlation between PP1A,GSDME and CD8+T cells abundance.Results The GEPIA database search showed that mRNA expression of PP1A and GSDME in colorectal cancer differed compared to normal tissues(P＜0.05).Western blot analysis showed that the relative expression of PP1A in colorectal cancer tissue was significantly higher than that in para-cancerous tissues(0.937 vs 0.643,P＜0.001).Immunohistochemical results showed that the expression of PP1A in colorectal cancer tissues was signifi-cantly higher than that in normal mucosa(P＜0.05).The ex-pression of GSDME in cancer tissue was closely correlated with patients'age,clinical stage and mismatch repair proteins(P＜0.05),and the distribution of CD8+T cells in the cancer infil-tration front was significantly higher than that in the normal mu-cosa,and the distribution of CD8+T cells in the cancer was cor-related with pT stage,clinical stage and lymph node metastasis.Spearman correlation analysis showed that PP1A was negatively correlated with GSDME expression(r=-0.196,P＜0.05).The overall survival PP1A-positive colorectal cancer patients was worse than that of PP1A-negative patients(P＜0.05),and the prognosis of patients was correlated with the degree of differentia-tion,lymph node metastasis,pT stage and clinical stage.Posi-tive expression of PP1A,degree of differentiation,clinical stage,pT stage and lymph node metastasis are independent risk factors affecting the prognosis of colorectal cancer patients.Conclusion PP1A is highly expressed in colorectal cancer and negatively correlated with GSDME-mediated cell pyroptosis,and the differ-ential expression of both is closely related to the progression and prognosis of colorectal cancer,which can be used as a potential indicator for judgment of the prognosis of colorectal cancer pa-tients.The differential distribution of CD8+T cells may be asso-ciated with GSDME-mediated cell pyroptosis and tumor develop-ment.

Objective:To investigate the surgical situations and perioperative outcome of pancreaticoduodenectomy in Jiangsu Province and the influencing factors for postoperative 90-day mortality.Methods:The retrospective case-control study was conducted. The clinicopathological data of 2 886 patients who underwent pancreaticoduodenectomy in 21 large tertiary hospitals of Jiangsu Quality Control Center for Pancreatic Diseases, including The First Affiliated Hospital of Nanjing Medical University, from March 2021 to December 2022 were collected. There were 1 732 males and 1 154 females, aged 65(57,71)years. Under the framework of the Jiangsu Provincial Pancreatic Disease Quality Control Project, the Jiangsu Quality Control Center for Pancreatic Diseases adopted a multi-center registration research method to establish a provincial electronic database for pancrea-ticoduodenectomy. Observation indicators: (1) clinical characteristics; (2) intraoperative and post-operative conditions; (3) influencing factors for 90-day mortality after pancreaticoduodenectomy. Measurement data with skewed distribution were represented as M( Q1, Q3) or M(IQR), and comparison between groups was conducted using the Mann-Whitney U test. Count data were expressed as absolute numbers or constituent ratio, and comparison between groups was conducted using the chi-square test, continuity correction chi-square test and Fisher exact probability. Maximal Youden index method was used to determine the cutoff value of continuous variables. Univariate analysis was performed using the corresponding statistical methods based on data types. Multivariate analysis was performed using the Logistic multiple regression model. Results:(1) Clinical characteristics. Of the 2 886 patients who underwent pancreaticoduodenectomy, there were 1 175 and 1 711 cases in 2021 and 2022, respectively. Of the 21 hospitals, 8 hospitals had an average annual surgical volume of <36 cases for pancreaticoduodenectomy, 10 hospitals had an average annual surgical volume of 36-119 cases, and 3 hospitals had an average annual surgical volume of ≥120 cases. There were 2 584 cases performed pancreaticoduodenectomy in thirteen hospitals with an average annual surgical volume of ≥36 cases, accounting for 89.536%(2 584/2 886)of the total cases. There were 1 357 cases performed pancrea-ticoduodenectomy in three hospitals with an average annual surgical volume of ≥120 cases, accounting for 47.020%(1 357/2 886) of the total cases. (2) Intraoperative and postoperative conditions. Of the 2 886 patients, the surgical approach was open surgery in 2 397 cases, minimally invasive surgery in 488 cases, and it is unknown in 1 case. The pylorus was preserved in 871 cases, not preserved in 1 952 cases, and it is unknown in 63 cases. Combined organ resection was performed in 305 cases (including vascular resection in 209 cases), not combined organ resection in 2 579 cases, and it is unknown in 2 cases. The operation time of 2 885 patients was 290(115)minutes, the volume of intra-operative blood loss of 2 882 patients was 240(250)mL, and the intraoperative blood transfusion rate of 2 880 patients was 27.153%(782/2 880). Of the 2 886 patients, the invasive treatment rate was 11.342%(327/2 883), the unplanned Intensive Care Unit (ICU) treatment rate was 3.087%(89/2 883), the reoperation rate was 1.590%(45/2 830), the duration of postoperative hospital stay was 17(11)days, the hospitalization mortality rate was 0.798%(23/2 882), and the failure rate of rescue data in 2 083 cases with severe complications was 6.529%(19/291). There were 2 477 patients receiving postoperative 90-day follow-up, with the 90-day mortality of 2.705%(67/2477). The total incidence rate of complication in 2 886 patients was 58.997%(1 423/2 412). The incidence rate of severe complication was 13.970%(291/2 083). The comprehensive complication index was 8.7(22.6) in 2 078 patients. (3) Influencing factors for 90-day mortality after pancreaticoduodenectomy. Results of multivariate analysis showed that age ≥ 70 years, postoperative invasive treatment, and unplanned ICU treatment were independent risk factors for 90-day mortality after pancreaticoduodenectomy ( odds ratio=2.403, 2.609, 16.141, 95% confidence interval as 1.281-4.510, 1.298-5.244, 7.119-36.596, P<0.05). Average annual surgical volume ≥36 cases in the hospital was an independent protective factor for 90-day mortality after pancreaticoduodenectomy ( odds ratio=0.368, 95% confidence interval as 0.168-0.808, P<0.05). Conclusions:Pancreaticoduodenectomy in Jiangsu Province is highly con-centrated in some hospitals, with a high incidence of postoperative complications, and the risk of postoperative 90-day mortality is significant higher than that of hospitallization mortality. Age ≥ 70 years, postoperative invasive treatment, and unplanned ICU treatment are independent risk factors for 90-day motality after pancreaticoduodenectomy, and average annual surgical volume ≥36 cases in the hospital is an independent protective factor.

In recent years, the high prevalence of myopia has become a global public health concern.Understanding the mechanism will help to achieve precise prevention and treatment of myopia.Animal models are important tools for studying the pathogenesis and prevention strategies.Mice are commonly used in myopia research, because they are easy to breed, feed and genetically manipulate.However, their small eye size makes it difficult to accurately measure the changes in biological parameters such as refraction and axial length.In addition, the refractive developmental pattern of mice is opposite to that of humans.This article reviews the structural features of the mouse cornea, lens, vitreous body, retina and sclera, the pattern of refractive development, and the latest progress about the establishment and evaluation of murine myopia models to provide some hints and references for further research.

OBJECTIVE To evaluate the mechanisms underlying the antidepressant effect of ZBH2012001,a novel serotonin and norepinephrine reuptake inhibitor(SNRI),in general and its ability to enhance monoaminergic transmission and suppress neuroinflammation in particular.METHODS① Male ICR mice were divided into vehicle(distilled water),duloxetine(DLX,10 or 20 mg·kg-1)and ZBH2012001(5,10 and 20 mg·kg-1)groups.One hour following ig administration,the antidepressant effect of ZBH2012001 was evaluated using the tail suspension test(TST)and forced swimming test(FST).② Radioligand binding assay was conducted to evaluate the affinity of ZBH2012001 for human serotonin transporters(hSERTs)and human norepinephrine transporters(hNETs).③ Mice were divided into vehicle(distilled water),DLX(10 or 20 mg·kg-1)and ZBH2012001(5,10 and 20 mg·kg-1)groups.One hour following drug administration,the 5-hydroxytryptophan(5-HTP)-induced head-twitch test or yohimbine-induced lethality test were performed to evaluate the effect of ZBH2012001 on the function of the 5-hydroxytryptamine(5-HT)and norepinephrine(NE)systems.④ Mice were divided into vehicle(distilled water+0.1%acetic acid),reserpine model(distilled water+reserpine 5 mg·kg-1),DLX(DLX 20 mg·kg-1+reserpine 5 mg·kg-1)and ZBH2012001(ZBH2012001 5,10 and 20 mg·kg-1+reserpine 5 mg·kg-1)groups.One hour following drug administration,reserpine was injected intraperitoneally to establish a monoamine-depletion model.The ptosis,akinesia,and hypothermia assays were performed to evaluate the effect of ZBH2012001 on the down-regulation of the reserpine-induced monoamine system.The TST in mice was used to evaluate the effect of ZBH2012001 on reserpine-induced depressive-like behavior while high-performance liquid chromatography with electrochemical detection(HPLC-ECD)was used to measure the levels of monoamines and their metabolites in the hippocampal tissue of reserpine-induced monoamine-depletion mice.ELISA was employed to detect the contents of tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)in the hippocampal tissue of reserpine-induced monoamine-depletion mice.Western blotting was used to assess the expressions of ionized calcium-binding adapter molecule-1(Iba-1)and nuclear factor-kappa B(NF-κB)in the hippocampal tissue of reserpine-induced monoamine-depletion mice.RESULTS ① Compared with the vehicle group,ZBH2012001(5,10 and 20 mg·kg-1)significantly reduced the immobility time both in the TST in mice(P<0.01,respectively),and ZBH2012001(20 mg·kg-1)and in the FST in mice(P<0.05).② ZBH2012001 competitively inhibited the binding of[3H]-imipramine to hSERTs and[3H]-nisoxetine to hNETs,with the half maximal inhibitory concentration(IC50)values of 84.95 and 712.90 nmol·L-1,respectively.③Com-pared with the vehicle group,ZBH2012001(10 and 20 mg·kg-1)significantly increased the head twitches induced by 5-HTP in mice(P<0.01,respectively)and increased the mortality rate in mice induced by yohimbine(P<0.05,P<0.01).④ In the reserpine-induced monoamine-depletion model in mice,compared with the vehicle group,mice in the reserpine model group exhibited ptosis,akinesia and hypothermia feature(P<0.01,respectively),significantly prolonged immobility time in the TST(P<0.01),significantly decreased the levels of NE,5-HT and dopamine(DA)(P<0.05,P<0.01),significantly increased the metabolic conversion rate of 5-HT and DA(P<0.01,respectively),significantly elevated levels of TNF-α and IL-6(P<0.05,respectively),and significantly increased expressions of Iba-1 and NF-κB(P<0.05,respectively)in the hippocampus.Compared with the model group,ZBH2012001(5,10 and 20 mg·kg-1)significantly antagonized ptosis and hypothermia behaviors induced by reserpine(P<0.01,respectively),ZBH2012001(10 and 20 mg·kg-1)significantly shortened the immobility time in reserpine-treated mice(P<0.05,P<0.01),ZBH2012001(20 mg·kg-1)significantly increased the levels of NE and 5-HT in the hippocampus of reserpine-treated mice(P<0.05,respectively),decreased the metabolic conversion rate of 5-HT(P<0.05),significantly reduced the contents of TNF-α and IL-6 in the hippocampus of reserpine-treated mice(P<0.05,respectively),ZBH2012001(5,10 and 20 mg·kg-1)significantly reduced the expression of Iba-1 protein in the hippocampus of reserpine-treated mice(P<0.01,respec-tively),and ZBH2012001(20 mg·kg-1)significantly reduced the expression of NF-κB protein in the hippocampus of reserpine-treated mice(P<0.05).CONCLUSION ZBH2012001 exerts its antidepres-sant effect through a dual mechanism involving monoamine enhancement and inflammation suppres-sion.

Objective To study the medication law of postoperative treatment of colorectal cancer by national TCM doctor Professor Piao Bingkui.Methods Professor Piao Bingkui's electronic medical records and paper medical records of colorectal cancer postoperative patients at Guang'anmen Hospital,China Academy of Chinese Medical Sciences and Beijing Yiqingtang Chinese Medicine Clinic were collected and organized from their outpatients from January 1st,2002 to February 28th,2022.R4.2.1 was used to study the prescriptions,including high-frequency drugs,drug types,properties of Chinese materia medica,yin yang and five elements and dosage of drugs,as well as the law of multi-drug association in postoperative patients with colorectal cancer.Results Totally 642 colorectal postoperative cancer patients were included,involving 2 226 prescriptions,180 kinds of Chinese materia medica,and a total frequency of 39 988 times.The high-frequency drugs were Astragali Radix,fried Aurantii Fructus with wheat bran,Dioscoreae Rhizoma,etc.The main drugs in terms of efficacy were tonics for tonifying deficiency,disinfectants,etc.;the main properties were warm and neutral,the main tastes were sweet,pungent and bitter,and the main meridians were spleen meridians and stomach meridians.Ascending medicines were often used,and the drug were basically non-toxic.The frequency of using yang tonifying medicine was high,and the five elements were commonly used as local medicines;the dosage was mostly 10,15 and 20 g.The complex network analysis and clustering analysis of the association between multiple drugs found that Professor Piao Bingkui's basic prescription for treating colorectal cancer included Astragali Radix,fried Aurantii Fructus with wheat bran,Dioscoreae Rhizoma,salt Alpiniae Oxyphyllae Fructus,Citri Reticulatae Pericarpium,Smilacis Glabrae Rhizoma,Pseudostellariae Radix,and fried Atractylodis Macrocephalae Rhizoma with wheat bran.Conclusion In the treatment of colorectal cancer,Professor Piao Bingkui focuses on reinforcing the healthy qi,nourishing spleen and stomach function,combined with detoxication method and clearing heat,expelling phlegm and dampness,guiding stagnation and dispelling stasis methods,making syndrome differentiation as well as tonifying and benefiting qi,regulating qi movement,so as to realize the"treating middle-energizer as balance"and achieve mild level when treating colorectal cancer.

Objective To evaluate the clinical efficacy of traditional Chinese medicine turbid phlegm obstructing lung decoction on patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD)and its influence on laboratory indexes.Methods A total of 191 patients with AECOPD who were hospitalized in the First People's Hospital of Changde from January 2021 to December 2022 were selected.Patients were divided into observation group(96 cases)and control group(95 cases)according to their treatment intention.The control group received conventional treatment of western medicine,and the observation group received oral administration of traditional Chinese medicine turbid phlegm obstructing lung decoction for one week.TCM symptom scores,COPD assessment test(CAT),lung function,laboratory indicators and efficacy were compared between two groups.Results The total effective rate of observation group was significantly higher than that of control group(χ2=4.573,P=0.030).After treatment,TCM symptom score,CAT score,hypersensitive C-reaction protein(hsCRP)and interleukin-6(IL-6)of patients in both groups were significantly lower than before treatment,percentage of forced vital capacity to predicted value(FVC%)and percentage of forced expiratory volume in one second to predicted value(FEV1%)were significantly higher than before treatment(P<0.05),arterial partial pressure of carbon dioxide(PaCO2)of observation group was lower than before treatment,and arterial partial pressure of oxygen(PaO2)was higher than before treatment(P<0.05).The TCM symptom score,CAT score,hsCRP and IL-6 of observation group were significantly lower than those of control group,while FVC%,FEV1%and PaO2 were significantly higher than those of control group(P<0.05).Conclusion On the basis of western medicine treatment,traditional Chinese medicine turbid phlegm obstructing lung decoction can more effectively improve clinical symptoms of AECOPD patients,relieve the inflammation in the body,contribute to the recovery of lung function and improve the quality of life of patients.