Objective:To construct the sphingosine kinase 1(SPHK1)overexpression lentiviral vector,and to establish the SKOV3 lentiviral stable transfection cell line.Methods:According to the SPHK1 data information provided by the National Center for Biotechnology Information(NCBI)database,the primers were designed and synthesized,the target gene was amplified,and connected to the GV492 plasmid treated with Bam HⅠ and AgeⅠ restriction enzymes to construct the SPHK1 overexpression lentiviral vector;the positive clones were selected for PCR and sequencing identification;the lentiviral plasmid and the lentiviral packaging auxiliary plasmid were co-transfected into the HEK-293T cells for packaging and titer determination;according to the measured optimal multiplicity of infection(MOI)of 10,the corresponding lentiviral amounts in various groups were transfected into the SKOV3 cells,and the SKOV3 cells were divided into blank group(without treatment),GV492 control group(GV492 control lentivirus infected SKOV3 cells),and GV492-SPHK1 overexpression group(GV492-SPHK1 overexpression lentivirus infected SKOV3 cells,ov-SPHK1 group);the optimal concentration of 2 mg·L-1 puromycin was used to screen the stably transfected SKOV3 cell line;after 48 h,the medium was changed and replaced with 1 mg·L-1 puromycin for screening for 14 d;the morphology and fluorescence expression of the cells were observed under fluorescence microscope;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of SPHK1 mRNA in the SKOV3 cells in various groups;Western blotting method was used to detect the expression level of SPHK1 protein in the SKOV3 cells in various groups.Results:The PCR sequencing results showed that the gene sequence of the SPHK1 overexpression lentiviral vector was completely consistent with the target sequence,and the SPHK1 overexpression lentiviral vector was successfully constructed;the titer determination results showed that the lentiviral titers in GV492 control group and ov-SPHK1 group were 5×1011 and 8×1011 TU·L?1,respectively;the SKOV3 cells in GV492 control group and ov-SPHK1 group were in good state and showed strong fluorescence expression,suggesting that the SKOV3 stable transfection cell line overexpressing SPHK1 was successfully established;the RT-qPCR results showed that compared with blank group and GV492 control group,the expression level of SPHK1 mRNA in the SKOV3 cells in ov-SPHK1 group was significantly increased(P<0.01);the Western blotting results showed that compared with blank group and GV492 control group,the expression level of SPHK1 protein in the SKOV3 cells in ov-SPHK1 group was significantly increased(P<0.01).Conclusion:The SPHK1 overexpression lentiviral vector is successfully constructed,and the SKOV3 stable transfection cell line is established.

Objective To use imaging features of pulmonary nodules to predict the risk of lymph node metastasis in patients with cT1-stage non-small cell lung cancer (NSCLC), providing a reference for clinical decision-making. Methods A retrospective analysis was conducted on the imaging features and postoperative pathological results of cT1 NSCLC patients who underwent surgical treatment at Linyi People’s Hospital from July 2019 to July 2022. Patients were grouped and analyzed according to lymph node metastasis status. Results A total of 1 123 patients were included, comprising 471 males and 652 females, with a median age of 59 (52, 66) years. Comparative analysis revealed that sex, age, nodule location, nodule size on imaging, solid component size, consolidation tumor ratio (CTR), average CT value, and tumor proximity to the pleura all influenced lymph node metastasis. A nomogram was constructed, indicating that the probability of lymph node metastasis in cT1 NSCLC was positively correlated with solid component size, CTR, and average CT value of the pulmonary nodule, and negatively correlated with patient age. The area under the receiver operating characteristic curve was 0.929. Conclusion For cT1 NSCLC patients, the probability of lymph node metastasis can be predicted by measuring the solid component size, CTR, and average CT value of the pulmonary nodule, in conjunction with patient age. However, relying solely on pulmonary nodule imaging characteristics is insufficient to determine a specific lymph node dissection strategy.

Objective To develop a symptoms assessment scale for lung transplant recipients in early postoperative period,and assess its reliability and validity so as to provide clinical staff with a tool to evaluate the symptoms in the patients in the early period after lung transplantation.Methods With a symptom experience model,a preliminary item pool was established through literature reviews,semi-structured interviews and expert panel meetings between December 2021 and January 2022.Based on the preliminary item pool,an initial scale was proposed after two rounds of expert consultation between February and March 2022.The initial scale was finalised after all items had revised for language expression based on the results of cognitive interviews conducted in March 2022.Subsequently,the final version of the scale was applied in the survey of 116 recipients of lung transplant in the early postoperative stage at Wuxi People's Hospital between April 2022 and February 2023.The reliability and validity of scale were then further tested.Results A total of 112 patients responded to the questionnaire survey.The developed scale comprised 5 dimensions:psychology-related symptoms,respiratory related symptoms,digestive related symptoms,circulatory related symptoms and other symptoms,with 18 symptom items in total.Content validity indexes of the Item-level content validity index(I-CVI))were 0.833 to 1.000 and the Scale-level content validity index(S-CVI)was at 0.954.Exploratory factor analysis revealed five common factors with a cumulative variance contribution rate of 82.366%.All the factors exhibited positive correlations with the scale,with the correlation coefficient at 0.760 to 0.837(P<0.01).The scale demonstrated a Cronbach α coefficient of 0.943,and the Cronbach α coefficients of the five common factors ranged from 0.869 to 0.941.Additionally,the scale exhibited a split-half reliability of 0.840,and the split-half reliability of the five common factors ranged from 0.830 to 0.937.Conclusions The symptoms assessment scale for lung transplant recipients in early postoperative period developed in this study exhibits good reliability and validity.It is feasible for evaluation of early symptoms in lung transplant recipients.

Antibody-drug conjugates(ADCs)are a new type of targeting antibodies that conjugate with highly toxic anticancer drugs via chemical linkers to exert high specificity and efficient killing of tumor cells,thereby attracting considerable attention in precise oncology therapy.Cetuximab(Cet)is a typical antibody that offers the benefits of good targeting and safety for individuals with advanced and inoperable cutaneous squamous cell carcinoma(cSCC);however,its anti-tumor activity is limited to a single use.Cisplatin(CisPt)shows good curative effects;however,its adverse effects and non-tumor-targeting ability are major drawbacks.In this study,we designed and developed a new ADC based on a new cytotoxic platinum(Ⅳ)prodrug(C8Pt(Ⅳ))and Cet.The so-called antibody-platinum(Ⅳ)prodrugs conjugates,named Cet-C8Pt(Ⅳ),showed excellent tumor targeting in cSCC.Specifically,it accurately delivered C8Pt(Ⅳ)into tumor cells to exert the combined anti-tumor effect of Cet and CisPt.Herein,metabolomic analysis showed that Cet-C8Pt(Ⅳ)promoted cellular apoptosis and increased DNA damage in cSCC cells by affecting the vitamin B6 metabolic pathway in tumor cells,thereby further enhancing the tumor-killing ability and providing a new strategy for clinical cancer treatment using antibody-platinum(Ⅳ)prodrugs conjugates.

Objective:To conduct a scoping review of the characteristics, functions, risk of bias and applications of home-based self-management behavior assessment tools for lung transplant patients, so as to provide references for clinical medical staff to conduct further related research.Methods:The relevant literatures were systematically searched in China National Knowledge Infrastructure, Wanfang Database, VIP, China Biology Medicine disc, PubMed, Web of Science Core Collection, Embase, Cochrane Library and CINAHL databases. The search period was from the establishment of the databases to June 30, 2023. The scoping review method framework was used to screen the literature, extract information and standardize the report.Results:A total of 10 167 Chinese and English literatures were searched, and after deduplication and screening, 23 articles that met the criteria were finally included, involving 13 home-based self-management behavior assessment tools for lung transplant patients, including nine specific tools and four universal tools. There were eight kinds of lung transplantation patients whose reliability and validity were not verified. Of the 13 tools, those with single-dimensional assessments and single measures predominate. The level of home self-management behavior of lung transplant patients was reflected mainly through compliance, and medication management was the most frequently assessed content.Conclusions:At present, the measurement of home-based self-management behavior of lung transplant patients is mainly based on questionnaire surveys, and the evaluation tools are mostly single dimensional. The reliability and validity of most evaluation tools in the population of lung transplant patients are not yet clear, and there is a certain risk of bias. It is recommended that researchers pay attention to the reliability and validity verification report of the evaluation tool in the target population when selecting it, and integrate multiple measurement methods to reduce measurement errors.

This paper summarized the research progress of palliative care for lung transplantation patients, to summarize the necessity and influencing factors of palliative care for lung transplantation patients, and defined the research status of palliative care implementation mode for lung transplantation patients, with a view to providing ideas for further research of palliative care on lung transplantation patients in China.

Objective    To investigate the feasibility and safety of hybrid coronary revascularization (HCR) in patients with multivessel coronary artery disease (MVCAD). Methods    A total of 50 patients with MVCAD who underwent HCR technique in our heart center from May 2016 to April 2019 were included in this study (a HCR group), including 38 patients who underwent two-stage HCR and 12 patients one-stop HCR. There were 39 males and 11 females, with an average age of 62.4 (46-82) years. Another 482 patients who underwent conventional median incision under off-pump coronary artery bypass grafting (OPCAB) at the same period were selected as control (an OPCAB group), including 392 males and 90 females, with an average age of 64.2 (48-84) years. The safety and feasibility of HCR were evaluated and compared with conventional OPCAB technique. Results    There was no perioperative death in both groups. Compared with the OPCAB, HCR was associated with shorter operation time, less chest tube drainage, lower requirement of blood transfusion, shorter mechanical ventilation time and shorter postoperative intensive care unit (ICU) stay (P<0.05). There was no statistical difference in the incidence of major adverse cardiac or cerebrovascular events during the follow-up of 6 to 36 months between the two groups. Conclusion    HCR provides favorable short and mid-term outcomes for selected patients with MVCAD compared with conventional OPCAB.

Microbial oils are potential resources of fuels and food oils in the future. In recent years, with the rapid development of systems biology technology, understanding the physiological metabolism and lipid accumulation characteristics of oleaginous microorganisms from a global perspective has become a research focus. As an important tool for systems biology research, omics technology has been widely used to reveal the mechanism of high-efficiency production of oils by oleaginous microorganisms. This provides a basis for rational genetic modification and fermentation process control of oleaginous microorganisms. In this article, we summarize the application of omics technology in oleaginous microorganisms, introduced the commonly used sample pre-processing and data analysis methods for omics analysis of oleaginous microorganisms, reviewe the researches for revealing the mechanism of efficient lipid production by oleaginous microorganisms based on omics technologies including genomics, transcriptomics, proteomics (modification) and metabolomics (lipidomics), as well as mathematical models based on omics data. The future development and application of omics technology for microbial oil production are also proposed.
Fermentation ; Lipids ; Metabolomics ; Proteomics ; Technology

Objective:To investigate the local infiltration of tissue-resident memory CD4 + T (CD4 + T RM) cells in lesions of patients with pemphigus and its clinical implications. Methods:From September 2017 to December 2018, 20 patients with pemphigus and 15 healthy human controls were collected from Department of Dermatology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine. Flow cytometry was performed to determine the proportion of CD4 + T RM cells in skin lesions of pemphigus patients and normal skin of healthy controls. The degree of CD4 + T RM cell infiltration in skin lesions was compared among different body sites of the patients with pemphigus, and the correlation between the proportion of CD4 + T RM cells and the time to disease control was analyzed. Normally distributed data were analyzed by using t test, and non-normally distributed data by using non-parametric test; the Pearson correlation coefficient was used to analyze correlations of the proportion of CD4 + T RM cells with pemphigus disease area index (PDAI) scores and circulating anti-desmoglein (Dsg) antibody titers. Results:Among the 20 patients, there were 16 with pemphigus vulgaris and 4 with pemphigus foliaceus. All the patients had skin involvement, 14 lesional tissue samples were taken from the trunk, and 6 from the limbs. There was no significant difference between the healthy control group and pemphigus group in terms of age, gender or biopsy sites (all P > 0.05) . The proportions of CD3 + T cells (72.75% ± 8.22%) and CD4 + T RM cells (44.05% ± 14.27%) in the skin lesions of patients with pemphigus were significantly higher than those in the skin tissues of the healthy controls (31.33% ± 8.72%, 12.60% ± 5.12%, t = 14.24, 9.10, respectively, both P < 0.001) . Among the patients with pemphigus, the proportion of CD4 + T RM cells was significantly higher in the skin lesions on the trunk (49.57% ± 12.32%) than in those on the limbs (31.17% ± 9.75%, t = 3.23, P < 0.05) . The proportion of CD4 + T RM cells in the skin lesions was positively correlated with the PDAI scores ( r2 = 0.246, P = 0.026) , but not correlated with serum titers of circulating anti-Dsg1 ( r2 = 0.137, P > 0.05) or anti-Dsg3 ( r2 = 0.162, P > 0.05) antibodies in the patients. During the treatment with systemic glucocorticoids, the proportion of CD4 + T RM cells in the skin lesions was significantly higher in the patients whose lesions could not be controlled within 4 weeks than in those whose lesions could be controlled within 4 weeks ( t = 3.22, P < 0.05) . Conclusion:The proportion of CD4 + T RM cells markedly increased in the skin lesions of patients with pemphigus, which may be related to the severity of the disease and response to treatment.

Objective To investigate the relationship between HBV -DNA load and serum markers in chronic hepatitis B( CHB) patients in Hohhot,Inner Mongolia,and to explore the mutation of HBV genotype and nucleoside analogue.Methods From January 2015 to December 2017,one hundred and ninety-three CHB patients hospitalized in the People＇s Hospital of Inner Mongolia were selected randomly.The clinical diagnostic criteria for all admitted patients were based on the " Guidelines for the Prevention and Treatment of Chronic Hepatitis B" jointly formulated by the Infectious Diseases Society of 2010. The HBV -DNA load of HBV was detected by real -time quantitative PCR,and the correlation between HBV -DNA load and serum markers was analyzed. Seventy -nine patients were selected from 193 hospitalized patients,PCR-reverse dot blot hybridization was used to analyze HBV genotyping and the drug resistance mutations of different genotypes.Results The differences of HBeAb level and HBV-DNA load between HBeAg positive patients and negative patients were statistically significant(all P<0.001). Of 79 serum specimens of HBV infected people,9 cases(11.4% ) were B genotypes,and 70 cases of C genotype (88.6% ).Of them,25 cases had different loci variation,the rate of variation was 31.6% (25/79),with the unit point rtS213T mutation dominated,accounting for about 24.0% (6/25).Conclusion In Hohhot Inner Mongolia patients with CHB,HBV-DNA load with HBeAg and HBe Ab level are correlated;genotype in patients including B type and C type,which is mainly genotype C;patients with CHB mainly had drug resistance to lamivudine and adefovir dipivoxil, mutations including rtS213T,and hybrid mutation.