Objective:To analyze the reoperation rate and causes during the early adoption phase of unilateral biportal endoscopy (UBE).Methods:The clinical data of 180 patients who underwent UBE performed by a single surgeon at Beijing Friendship Hospital, Capital Medical University from October 2021 to June 2023 were retrospectively analyzed. Clinical and imaging data of patients who underwent reoperation were collected to analyze the causes of reoperation, and the clinical efficacy of the reoperations was also followed up. Measurement data were expressed as mean ± standard deviation ( ± s), and t-test was used before and after treatment. Results:A total of 180 patients who underwent UBE were included in this study, of which 6 patients underwent reoperation, and the reoperation rate was 3.33%. Among them, 3 cases occurred in the first 90 surgeries and the other 3 occurred in the subsequent 90 surgeries. The causes of reoperation were as follows: recurrent lumbar disc herniation at the same segment postoperatively in 2 cases, insufficient decompression in 2 cases, disc herniation following isolated decompression in 1 case, and immediate postoperative perianal numbness in 1 case. The time between the initial surgery and reoperation ranged from 0 to 187 days, with an average of 63.3 days. The average follow-up time after reoperation was 18.3 months. The visual analogue scale (VAS) and Oswestry disability index (ODI) scores of the patients at the last follow-up were significantly improved compared with those before operation (VAS score of low back pain: 5.2 ± 1.7 before operation, 1.2 ± 0.8 at the last follow-up, P<0.001; VAS score of leg pain: 7.2 ± 1.5 before operation, 1.2 ± 1.2 at the last follow-up, P<0.001; ODI score: 67.3 ± 5.7 before operation, 20.2 ± 8.2 at the last follow-up, P<0.001). The postoperative modified MacNab scores were generally satisfactory (4 cases were rated as excellent, accounting for 66.7%; 2 cases were rated as good, accounting for 33.3%). Except for one patient who experienced dural injury during open revision surgery, there were no serious complications such as nerve damage. Conclusions:In the early stages of UBE surgery, recurrent lumbar disc herniation and inadequate decompression are the primary reasons for reoperation, typically occurring within the first three months postoperatively. Reoperation does not significantly increase the risk of nerve injury. Enhanced early postoperative follow-up is recommended. For symptomatic patients, a second surgery with thorough decompression can yield satisfactory treatment outcomes.

Objective To construct recombinant human butyrylcholinesterase(rhBChE)knock-in goat fetal fibroblast cell lines(GFFs)by using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)-mediated homology-directed repair mechanism for subsequent production of goat expressing rhBChE.Methods The efficient sgRNA sites targeting goat[3-casein(CSN2)gene were designed and screened,and the targeting efficiency of the sgRNA in goat mammary epithelial cells(GMECs)was confirmed by electro-transfection,flow sorting,and sequencing of PCR products.The red fluorescent reporter gene homology repair vector(P2A-mCherry)targeting the sgRNA was constructed,and then the integration and expression efficiency was detected by flow cytometry.The rhBChE homology repair vector(P2A-rhBChE)targeting the sgRNA of CSN2 gene was constructed in GFFs,the rhBChE positive cell clones were obtained via electro-transfection and flow sorting,and the rhBChE knock-in cell lines was identified by sequencing of PCR products.Results The sgRNA4 was identified as an efficient target of goat CSN2 gene,which could be also used for targeted integration of other genes.Three rhBChE knock-in cell lines were successfully constructed.Conclusion The rhBChE knock-in GFFs targeting goat CSN2 gene lays the foundation for the production of mammary bioreactors expressing rhBChE.

Objective To explore the sufentanil by adjusting the adenosine monosodium phosphate activa-ted protein kinase(AMPK)/thioredoxin interacting proteins(TXNIP)/nucleotide-binding oligomerization do-main-like receptor protein 3(NLRP3)signaling pathways of the alveolar epithelial cells induced by lipopo-lysaccharide to the dead.Methods Human alveolar epithelial type Ⅱ cell(AECⅡ)were cultured to logarith-mic phase.Then they were divided into control group(normal glucose culture),lipopolysaccharide group(li-popolysaccharide induced injury by 50 μg/mL),low concentration sufentanil group(20 μmol/L),high concen-tration sufentanil group(40 μmol/L),high concentration sufentanil(40 μmol/L)+AMPK-IN-3 group(50μmol/L AMPK inhibitors).MTT assay was used to detect cell proliferation.Pyroptosis was detected by Ho-echst 33342 and propidium iodide(PI)double staining.The levels of interleukin(IL)-1β,IL-18 and tumor nec-rosis factor-α(TNF-α)in cell supernatant were detected by enzyme-linked immunosorbent assay.Quantitative real-time PCR was used to detect the mRNA expression of focal death related factors cysteinyl proteinase-1(Caspase-1),apoptosis-associated speck-like protein(ASC),and gasdermin D(GSDMD).The protein expres-sion of AMPK,phosphorylated adenosine monosodium phosphate activated protein kinase(p-AMPK),TXNIP and NLRP3 were detected by Western blot.Results Compared with the control group,the survival rate of AEC Ⅱ cells in the lipopolysaccharide group decreased,the number of PI+positive cells increased,the release of inflammatory factors IL-1β,IL-18 and TNF-α increased,the expression of Caspase-1,ASC and GSDMD mR-NA increased,the expression of p-AMPK/AMPK decreased,and the expression of TXNIP and NLRP3 in-creased(P＜0.05).Compared with the lipopolysaccharide group,the survival rate of AEC Ⅱ cells in the low concentration sufentanil group and the high concentration sufentanil group increased,PI+positive cells de-creased,the release of inflammatory factors IL-1β,IL-18 and TNF-α decreased,the expression of Caspase-1,ASC and GSDMD mRNA decreased,the expression of p-AMPK/AMPK increased,and the expression of TX-NIP and NLRP3 decreased(P＜0.05).Compared with the high concentration sufentanil group,the survival rate of AEC Ⅱ cells in the high concentration sufentanil+AMPK-IN-3 group decreased,the number of PI+positive cells increased,the release of inflammatory factors IL-1β,IL-18 and TNF-α increased,the expression of Caspase-1,ASC and GSDMD mRNA increased,the expression of p-AMPK/AMPK decreased,and the ex-pression of TXNIP and NLRP3 increased(P＜0.05).Conclusion Sufentanil may improve lipopolysaccharide induced alveolar epithelial cell pyroptosis by regulating AMPK/TXNIP/NLRP3 signaling pathway.

In recent years， clinical studies on targeted therapy and immunotherapy for advanced hepatocellular carcinoma used alone or in combination have provided abundant evidence on efficacy and safety for the selection of first-line therapies. However， no consensus has been reached on the selection of second-line therapies in various clinical guidelines for hepatocellular carcinoma， which is caused by the fact that existing evidence is limited to the options after failure of sorafenib and that there is still a lack of high-level evidence for new first-line therapies such as second-line therapies after resistance to targeted therapy and immunotherapy for hepatocellular carcinoma. This article reviews the results of current clinical trials and summarizes the studies on second-line therapies for hepatocellular carcinoma after resistance to first-line targeted therapy and immunotherapy for hepatocellular carcinoma based on the different mechanisms of action of drugs， as well as the research advances in recent years. For hepatocellular carcinoma patients with resistance to first-line targeted therapy and immunotherapy， targeted combination therapy and dual-immune therapy are expected to improve treatment outcome and survival， and more prospective clinical studies are needed in the future to provide effective and safe treatment regimens for hepatocellular carcinoma patients with resistance to targeted therapy and immunotherapy.

AIM:To explore the effects of CD38 on lysosome reformation and cholesterol efflux in macro-phages.METHODS:Bone marrow-derived macrophages from low-density lipoprotein(LDL)receptor knockout(LDLr-/-)mice were cultured as cell model.Live cell imaging system was applied to evaluate the effect of nicotinic acid adenine di-nucleotide phosphate(NAADP)on lysosome number.ELISA was conducted to measure NAADP level in macrophages.After the cells were treated with nicotinic acid(NA),RT-qPCR was conducted to detect CD38 mRNA expression,and Western blot was conducted to observe CD38 protein expression and phosphorylated transcription factor EB(TFEB)level.Laser scanning confocal microscopy was applied to evaluate the influence of CD38/NAADP signaling on lysosome number and cholesterol egression.RESULTS:NAADP remarkably increased lysosome number(P<0.05),and this effect was significantly inhibited by NAADP antagonist NED-19,Ca2+ chelator BAPTA,and calcineurin inhibitor CsA(P<0.05).CD38 markedly enhanced NAADP synthesis in macrophages(P<0.05).NAADP synthetic substrate NA prominently ele-vated the expression of CD38 mRNA and protein(P<0.05).NA significantly decreased the phosphorylated TFEB level;this effect was also attenuated by NED-19,BAPTA and CsA(P<0.05).Disrupting CD38/NAADP signaling pathway markedly inhibited NA-induced enhancement of lysosome number,lysosomal free cholesterol and cytosol cholesterol ester efflux in macrophages(P<0.05).NA-induced enhancement of lysosome number,lysosomal free cholesterol and cytosol cholesterol ester efflux abolished in LDLr/CD38 DKO macrophages(P<0.05),whereas these effects induced by NA were recovered after CD38 gene rescue.CONCLUSION:CD38 triggers lysosome reformation via TFEB and consequently pro-motes the efflux of lysosomal free cholesterol and cytosol cholesterol ester.

Objective:To implement scoping review of the quality evaluation tool for mobile medical applications (mHealth APP) based on the perspective of health promotion, so as to provide reference for the development or introduction of tools suitable for China and provide inspiration for conducting high quality mHealth APP electronic health intervention research.Methods:Following the method of scoping review, mHealth APP quality evaluation tool development and validation articles published from January 2008 to February 2022 were systematically searched in Cochrane Library, PubMed, EMbase, Institute for Scientific Information (ISI) Web of Science, SCOPUS, CINAHL, PsyINFO, China National Knowledge Infrastructure, VIP, WanFang Data and China Biomedical Literature Database. This study extracted the basic information of the tool and used topic analysis to summarize and integrate the content of the tool items.Results:A total of 27 studies were included, involving 14 mHealth APP quality evaluation tools. The theme analysis process of evaluating the content of items generated a total of 300 primary codes, which were classified into four themes, including software attributes, medical science, behavioral science, and user experience.Conclusions:As a cross disciplinary product, the mHealth app faces multiple challenges in terms of quality. At present, the focus of existing quality evaluation tools varies, among which scientific evaluation originates from the special positioning of mHealth APP and needs to be taken seriously. Different evaluation perspectives may be potential factors that affect the quality evaluation results, and as a key link, users' needs need to be further focused and explored.

Objective:To compare the safety and efficacy of the " step-up approach" versus the " step-jump approach" in treatment of infected pancreatic necrosis (IPN).Method:The clinical data of IPN patients who underwent step-up strategy or step-jump strategy treatment at the Department of Pancreatic and Biliary Surgery of the First Affiliated Hospital of Harbin Medical University from December 2018 to November 2022 were analyzed retrospectively. Propensity score matching (PSM) was done based on the nearest neighbor matching method (1: 1 ratio). After matching the baseline data (the caliper value was 0.01), a total of 62 patients with IPN were included, including 41 males and 21 females, aged (41.1±13.1) years old. Patients who were treated with the step-up strategy were included in the step-up group, while patients who were treated with the step-jump strategy were included in the step-jump group. There were 31 patients in each group after PSM, and the treatment effect of the two groups were compared.Results:Of the 62 patients with IPN, 43 received surgical intervention, and 19 were managed successfully using symptomatic anti-inflammatory treatment or percutaneous catheter drainage. The total hospitalization cost of patients in the step-jump group was significantly higher than that in the step-up group [122 000 (73 000, 179 000) yuan vs. 88 000 (46 000, 144 000) yuan, P=0.034]. The overall cure rate of IPN patients in the step-jump group was 93.5%(29/31). The 2 patients who died had type Ⅲ IPN. In the IPN patients in the step-up group were all cured, and the overall cure rate was 100%(31/31), with no death. There were no statistical differences between the two groups in the rates of death, postoperative complications, residual infection, debridement ≥2 times, and positive bacterial culture in blood or drainage fluid (all P>0.05). A total of 19.4% (12/62) patients had postoperative complications, including 4 patients with abdominal bleeding, 3 patients with new organ dysfunction, 2 patients with gastrointestinal bleeding, 2 patients with gastrointestinal fistula, and 1 patient with venous thrombosis in both lower limbs. Conclusion:Both the step-up treatment strategy and the step-jump treatment strategy were safe and effective for treatment of IPN patients.

Objective:To study the serological and molecular biological characteristics and differences of occult hepatitis B infection (OBI) between blood donors and chronic hepatitis B (CHB) patients.Methods:Nineteen OBI samples from blood donors and Nineteen OBI samples from CHB patients were collected, and named as group A and group B, respectively. Chemiluminescence method was used for hepatitis B five items detection. Real-time PCR method was used for HBV DNA quantification, and S gene was amplified by nested PCR and sequenced by DNA sequencing.Results:The level of HBV DNA in group A was significantly lower than that in group B ( P<0.05), and there were no significant differences in the serological result between the two groups. The mutation rate in the " a" determinant of S gene in group B was significantly higher than that in group A ( P<0.05). Conclusions:Compared with CHB patients, OBI blood donors have lower levels of viral replication and less chance of S gene mutation, and the use of highly sensitive nucleic acid detection reagents and method during blood screening can maximize the safety of blood transfusion.

Objective: To investigate the risk factors for yersiniosis, so as to provide insights into prevention of yersiniosis. Methods: The patients with yersiniosis admitted to the clinics in the surveillance site of Chengbei Township of Jin'an District and Chengnan Township of Yu'an District in Lu'an City from 2013 to 2021 were included as the case group, and the healthy family members matched to cases were selected as the family control group, while normal residents with a 1︰2 match in the same village, gender, and age difference within 5 years were included in the community control group. Participants' demographics, hand-washing and eating habits, living environment hygiene, poultry and livestock feeding were collected using questionnaire surveys, and factors affecting yersiniosis were identified using a multivariable conditional logistic regression model. Results: There were 43 cases in the case group, with a median (interquartile range) age of 45 (34) years, 91 cases in the family control group, with a median (interquartile range) age of 36 (36) years and 86 cases in the community control group, with a median (interquartile range) age of 46 (34) years. Multivariable conditional logistic regression analysis showed that compared with the family control group, the habit of drinking unboiled water (OR=6.721, 95%CI: 1.765-25.588), and direct consumption of food stored in the refrigerator (OR=7.089, 95%CI: 1.873-26.829) were risk factors for yersiniosis in the case group; and compared with the community control group, not washing hands after contacting with poultry and livestock (OR=50.592, 95%CI: 2.758-927.997), habit of eating raw vegetables and fruits (OR=5.340, 95%CI: 1.022-27.887), direct consumption of food stored in the refrigerator (OR=19.973, 95%CI: 2.118-188.336), and unclean refrigerator (OR=12.692, 95%CI: 1.992-80.869) were risk factors for yersiniosis in the case group. Compared with the family and community control groups, not washing hands after contacting with poultry and livestock (OR=4.075, 95%CI: 1.427-11.637), habit of drinking unboiled water (OR=4.153, 95%CI: 1.331-12.957), habit of eating raw vegetables and fruits (OR=4.744, 95%CI: 1.609-13.993), and direct consumption of food stored in the refrigerator (OR=5.051, 95%CI: 1.773-14.395) were risk factors for yersiniosis in the control group. Conclusion Unhealthy habits such as eating raw vegetables and fruits, drinking unboiled water, direct consumption of food stored in the refrigerator, unclean refrigerator, and not washing hands after contacting poultry and livestock may increase the risk of yersiniosis.