[摘 要] 目的：探讨溶酶体相关膜蛋白3（LAMP3）与转录激活因子4（ATF4）在宫颈癌中的表达及其与临床病理参数的相关性。方法：采用免疫组化SP法检测正常宫颈组织、宫颈上皮内病变组织及宫颈癌组织中LAMP3与ATF4的表达情况，分析两种蛋白与宫颈癌患者临床病理参数之间的关系，并且分析两种蛋白的相关性。结果：LAMP3在正常宫颈组及高级别鳞状上皮内病变组中均为阴性或低表达，在宫颈癌中高表达率为38.3%，差异有统计学意义（χ2 = 14.113，P = 0.001）。ATF4在正常宫颈组中高表达率为26.7%，在高级别鳞状上皮内病变组中高表达率为10.0%，在宫颈癌组中高表达率为58.3%，差异有统计学意义（χ2 = 11.078，P = 0.004）。LAMP3的表达与宫颈癌FIGO分期（χ2 = 10.139，P = 0.006）、淋巴结转移（χ2 = 8.475，P = 0.004）有关，差异均有统计学意义。ATF4的表达在宫颈癌病灶大小（χ2 = 4.578，P = 0.032）、FIGO分期（χ2 = 8.971，P = 0.009）、淋巴结转移（χ2 = 7.881，P = 0.005）等方面的差异均有统计学意义。LAMP3与ATF4在宫颈癌中的表达呈正相关（r = 0.388，P = 0.002）。结论：LAMP3与ATF4在宫颈癌组织中表达升高，两者的表达程度具有相关性，且在宫颈癌的发生发展中发挥重要的促进作用，有望成为宫颈癌治疗的潜在靶点。

Atherosclerosis is a complex pathological condition resulting from lipid deposition， chronic inflammatory responses， and fibrosis， with a prolonged disease course and multifactorial etiology. Based on the traditional Chinese medicine （TCM） theory of accumulation syndrome， atherosclerosis can be classified under this category， with its pathogenesis involving phlegm， blood stasis， deficiency， and accumulation. This paper proposed a stage-based intervention strategy using the four therapeutic principles of "attacking， supplementing， dispersing， dissipating"， and divided into six stages based on the pathological progression， including the stage of accumulation before formation， the stage of accumulation already formed， the stage of nucleus accumulation， the stage of nucleus accumulation decay， the stage of nucleus accumulation consolidation， and the stage of severe stenosis of nucleus. At different stages， the intervention focuses on reinforcing healthy qi and consolidating the root， tonifying the kidneys and spleen， dispersing and removing turbidity， removing phlegm stagnation， promoting qi circulation， dispersing accumulations and removing stasis， attacking accumulation and expelling stasis， directing the turbid downward and dispersing accumulation， and treatment would be adjusted based on specific symptoms， which provides a theoretical framework for the prevention and treatment of atherosclerosis with TCM.

TFIIB-related factor 1 (BRF1) is an important transcription factor. It specifically regulates the transcription of RNA polymerase III-dependent genes (RNA Pol III genes). The products of these genes are some small non-coding RNAs, including transfer RNAs (tRNAs) and 5S ribosomal RNAs (5S rRNA). The transcription levels of tRNAs and 5S rRNA vary with changes in intracellular BRF1 amounts. tRNAs and 5S rRNA play a crucial role in determining protein synthesis. Studies have demonstrated that dysregulation of tRNAs and 5S rRNA is closely related to cell growth, proliferation, transformation, and even tumorigenesis. BRF1 is a key factor determining the generation of tRNAs and 5S rRNA. Increasing BRF1 expression enhances cell proliferation and transformation, promoting tumor development. In contrast, repressing BRF1 activity decreases the rates of cell proliferation and transformation, and inhibits tumor growth. High levels of BRF1 are found in the samples of patients suffering from hepatocellular carcinoma, breast cancer, gastric carcinoma, lung cancer, prostate carcinoma, and other cancers. It indicates that high levels of BRF1 are closely related to the occurrence of human cancer and may be a common landmark of tumors. But there is discrepancy in the regulatory mechanisms and signaling pathways of BRF1 overexpression in different cancers. In general, high levels of BRF1 in patients suffering from cancer show short survival period and poor prognosis. However, there is one exception, namely breast cancer. Approximate 80% of cases of breast cancer are estrogen receptor-positive (ER+) and 20% are ER-. The cases with high levels of BRF1 reveal longer survival period and better prognosis after they accepted the hormone treatment by Tamoxifen (Tam), compared to the cases with low level BRF1. It seems like a contradiction. Most of the cases with high levels of BRF1 belong to ER+ status. Tam has been used to treat ER+ cases of breast cancer after diagnosis and surgery. Thus, hormone therapy, such as Tam, is more effective on these patients. This is because, on one hand, that Tam competes with E2 (17β-estradiol) to bind to estrogen receptor α (ERα), but does not dissociate to occupy the receptors, blocking E2 binding to this receptor and inhibiting its biological effects. On other hand, Tam can inhibit the expression of BRF1, leading to a decline of intracellular BRF1 levels. Therefore, the actual levels of BRF1 are lower in the patients with ER+ breast cancer. It appears the prognosis of the high BRF1 expression cases better than that of the low BRF1 expression cases. Myocardial hypertrophy manifests magnification of cardiomyocyte volume rather than number increasing in the postnatal heart. Myocardial hypertrophy is a critical risk factor underlying cardiovascular diseases. No matter how myocardial hypertrophy occur, it will ultimately lead to myocardial dysfunction and heart failure. Hypertrophic growth of cardiomyocytes requires a large amount of protein synthesis to meet its needs of cardiomyocyte growth. Animal models and cell experiments have shown that myocardial hypertrophy stimulates a significant increase in BRF1 expression and transcription of tRNAs and 5S rRNA. Interestingly, elevated levels of BRF1 are found in the myocardium tissues of patients with myocardial hypertrophy. These studies demonstrate that BRF1 indeed plays a critical role in myocardial hypertrophy. In summary, high levels of BRF1 are found in patients suffering from different cancers and myocardial hypertrophy. It implies that BRF1 is a promising biological target of cancer and cardiomyopathy. BRF1 is expected to become a common biomarker for early diagnosis and prognostic observation of different human cancers. It is also an important biomarker for the diagnosis and treatment of cardiomyopathy. BRF1 not only holds an important position in the field of basic medical research but also has great prospects for translational medicine. In the present article, we summarize the progress on studies of BRF1 expressions in cancer and cardiomyopathy, proposes future research directions. It is a new research area. Here, we emphasize the significancy of BRF overexpression in the two huge diseases of human, cancer and cardiomyopathy to raise people's attention to this field.

Background Existing studies have confirmed that fine particulate matter (PM_2.5)is one of the important factors inducing pulmonary fibrosis. Pulmonary fibrosis is the terminal stage of a major category of lung diseases characterized by the destruction of tissue structure, and eventually leading lung ventilation and ventilation dysfunction. No effective pulmonary fibrosis treatment is available yet. Objective To investigate the protective effect of salidroside on pulmonary fibrosis induced by the exposure of PM_2.5 and its molecular mechanism. Methods Seventy 7-week-old male C57BL/6 mice were randomly divided into four groups: control group (intratracheal instillation of normal saline + saline by gavage, n=25), Sal group (intratracheal instillation of normal saline + Sal 60 mg·kg⁻¹ by gavage, n=10), PM_2.5 group (intratracheal instillation of PM_2.5 5 mg·kg⁻¹ + saline by gavage, n=10), and Sal + PM_2.5 group (intratracheal instillation of PM_2.5 5 mg·kg⁻¹ +Sal 60 mg·kg⁻¹ by gavage, n=10). The mice were administered by gavage once daily, intratracheal instillation once every 3 d, and every 3 d constituted an experimental cycle. At the end of the 26-30th cycles, 3 mice in the control group and 3 mice in the PM_2.5 group were randomly sacrificed, and the lung tissues were collected for Masson staining to verify whether the pulmonary fibrosis model was successfully established. After 30 cycles, the model was successfully constructed. After 1 week of continuous observation, the mice were sacrificed, and the blood and lung tissues of the mice were collected to make lung tissue sections. Assay kits were correspondingly employed to detect oxidative stress indicators such as serum malondialdehyde (MDA) and superoxide dismutase (SOD). Western blotting was used to detect the expression of fibrosis-related proteins (Collagen-III, α-SMA), mitochondrial dynamics-related proteins (MFN1, Drp1), and mitophagy-related proteins (PINK1, Parkin, and LC3). Results Compared with the control group, the weight gain rate of the PM_2.5 group was slowed down (P<0.05), which was alleviated by the Sal intervention (P<0.05). The lung coefficient increased after the PM_2.5 exposure (P<0.05), which was alleviated by Sal intervention. Compared with the control group, the PM_2.5 group showed severe alveolar structure damage, inflammatory cell infiltration, and blue collagen deposition, and significantly increased the lung injury score, collagen volume fraction (CVF), Szapiel score, and Ashcroft score (P<0.05), as well as serum oxidative stress levels (P<0.05). The protein expression levels of Collagen-III, α-SMA, Drp1, PINK1, Parkin, and LC3 II/I were increased (P<0.05), and the expression of MFN1 was decreased (P<0.05). Compared with the PM_2.5 group, the Sal intervention alleviated lung injury, reduced inflammatory cell infiltration and collagen deposition, showing decreased lung injury score, CVF, Szapiel score, and Ashcroft score (P<0.05), and decreased serum oxidative stress levels (P<0.05); the protein expression levels of Collagen-III, α-SMA, PINK1, Parkin, and LC3 II/I were decreased (P<0.05), the expression level of Drp1 was decreased, and the expression level of MFN1 was increased. Conclusion In the process of pulmonary fibrosis induced by PM_2.5 exposure in mice, Sal may affect mitochondrial autophagy through PINK1/Parkin pathway and play a protective role. The specific mechanism needs to be further verified.

DNA damage triggers cells to initiate a series of DNA damage response(DDR),including DNA damage repair,cell cycle checkpoint activation,cell cycle arrest,activation of various intracellular signal transduction pathways,and cell apoptosis,etc.DNA Damage Repair,an important mechanism by which cells maintain genomic stability,was awarded the Nobel Prize in Chemistry in 2015.DNA damage repair pathways mainly include:base-excision repair(BER),nucleotide excision repair(NER),mis-match repair(MMR),homologous recombination(HR)and non-homologous end joining(NHEJ).They play an important role in the repair of DNA damage such as single-strand break(SSB)or double-strand break(DSB).DNA damage repair defects are closely related to tumorigenesis and development and is also an important target for tumor therapy.Poly-ADP-ribose polymerase(PARP)and breast canc-er susceptibility gene BRCA1/2 and others in the DNA damage repair pathways have synthetic lethality effects.It makes PARP inhibitor(PARPi)be the first and currently the only commercially available syn-thetic lethal target drug for tumor therapy.PARPi has good efficacy in the treatment of ovarian cancer and a variety of solid tumors,which make the research and development of synthetic lethal target drugs related to DNA damage repair and DDR pathway become a hot spot.Other targets under research mainly in-clude:ataxia telangiectasia-mutated protein(ATM),ataxia telangiectasia and RAD3 related protein(ATR),DNA-dependent protein kinase catalytic subunit(DNA-PKcs),checkpoint kinasel(CHK1),Checkpoint kinase 2(CHK2),mitogen-preventing protein kinase WEE 1,etc.The combination of PARPi with other DDR target drugs,anti-angiogenesis drugs or immune checkpoint inhibitors may be-come effective means and development prospect to overcome PARPi resistance and improve the therapeu-tic effect.Here we review the key molecules and potential tumor therapeutic targets in DNA damage re-pair and related DDR pathways,and the research on synthetic lethal targets and their application and prospect in ovarian cancer.We aim to provide guidance for basic research and clinical application.

The PCR-amplified severe fever with thrombocytopenia syndrome virus(SFTSV)Gn-DⅢ-Ⅲ gene was inserted into the pET-30a(+)prokaryotic expression vector to generate the re-combinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ.The plasmid was transformed into E.coli BL21(DE3)for Gn-DⅢ-m protein expression and the expression conditions were optimized.The Gn-DⅢ-Ⅲ protein purified with Ni-NTA column affinity chromatography was applied as the captured antigen to establish an indirect ELISA method for the detection of SFTSV antibody.The results demonstrated that the recombinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ was successfully constructed as identified by PCR and sequencing.The recombinant protein SFTSV Gn-D m-Ⅲ was soluble ex-pression in E.coli under the optimal induction conditions of 0.4 mmol/L IPTG at 25 ℃ for 4 h,and the protein purity was 91.77％after purification by Ni-NTA column.The optimal reaction con-ditions for the indirect ELISA of SFTSV antibody were as follows:coating antigen concentration(5 μg/mL),primary antibody(incubation at 37 ℃ for 1.5 h),and secondary antibody(diluted 1:10 000 and incubated at 37 ℃ for 1 h).The established method had no cross-reactivity with Rift Valley fever virus(RVFV),Ebola virus(EBOV),and tick-borne encephalitis virus(TBEV)posi-tive sera.The method had a high sensitivity,with P/N＞2.1 for SFTSV-positive sera diluted to 81920.Coefficients of variation for intra-and inter-batch reactions were less than 10％.Detection of four SFTSV-infected human clinical serum samples showed the serum samples from patients in re-mission were tested as positive(P/N＞2.1),while serum samples from patients with multiple or-gan failure were detected as negative(P/N＜2.1).The results indicated that the SFTSV Gn-D Ⅲ-Ⅲ protein was successfully expressed and purified,and it was used as the coating protein to estab-lish an indirect ELISA assay for SFTSV antibody,which possesses good specificity,sensitivity and reproducibility.This method might be applied to detect human SFTSV clinical serum samples.

Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-positive bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-positive bacteria from blood cultures in member hospitals of National Bloodstream Infection Bacterial Resistant Investigation Collaborative System（BRICS）were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute（CLSI）. WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:A total of 3 163 strains of Gram-positive pathogens were collected from 51 member units，and the top five bacteria were Staphylococcus aureus（ n=1 147，36.3%），coagulase-negative Staphylococci（ n=928，29.3%）， Enterococcus faecalis（ n=369，11.7%）， Enterococcus faecium（ n=296，9.4%）and alpha-hemolyticus Streptococci（ n=192，6.1%）. The detection rates of methicillin-resistant Staphylococcus aureus（MRSA）and methicillin-resistant coagulase-negative Staphylococci（MRCNS）were 26.4%（303/1 147）and 66.7%（619/928），respectively. No glycopeptide and daptomycin-resistant Staphylococci were detected. The sensitivity rates of Staphylococcus aureus to cefpirome，rifampin，compound sulfamethoxazole，linezolid，minocycline and tigecycline were all >95.0%. Enterococcus faecium was more prevalent than Enterococcus faecalis. The resistance rates of Enterococcus faecium to vancomycin and teicoplanin were both 0.5%（2/369），and no vancomycin-resistant Enterococcus faecium was detected. The detection rate of MRSA in southern China was significantly lower than that in other regions（ χ2=14.578， P=0.002），while the detection rate of MRCNS in northern China was significantly higher than that in other regions（ χ2=15.195， P=0.002）. The detection rates of MRSA and MRCNS in provincial hospitals were higher than those in municipal hospitals（ χ2=13.519 and 12.136， P<0.001）. The detection rates of MRSA and MRCNS in economically more advanced regions（per capita GDP≥92 059 Yuan in 2022）were higher than those in economically less advanced regions（per capita GDP<92 059 Yuan）（ χ2=9.969 and 7.606， P=0.002和0.006）. Conclusions:Among the Gram-positive pathogens causing bloodstream infections in China， Staphylococci is the most common while the MRSA incidence decreases continuously with time；the detection rate of Enterococcus faecium exceeds that of Enterococcus faecalis. The overall prevalence of vancomycin-resistant Enterococci is still at a low level. The composition ratio of Gram-positive pathogens and resistant profiles varies slightly across regions of China，with the prevalence of MRSA and MRCNS being more pronounced in provincial hospitals and areas with a per capita GDP≥92 059 yuan.

[Objective]To investigate the clinical effect of acupoint application combined with Xingpi Yang'er Granules in the treatment of children with dyspeptic diarrhea and the changes of gastrointestinal hormones.[Methods]One hundred and ten children with dyspeptic diarrhea treated in the Department of Pediatrics of Hengshui Maternal and Child Health Care Hospital from June 2022 to June 2023 were randomly divided into control group(55 cases)and observation group(55 cases).The control group was given Clostridium butyricum acid live powder,and the observation group was given Xingpi Yang'er Granules at the same time for point application,continuous treatment for 9 days.Traditional Chinese medicine(TCM)syndrome score,clinical efficacy,symptom improvement time,gastrointestinal indexes,including gastrin(GAS),motilin(MTL)and somatostatin(SS)and adverse reactions were compared between the two groups.[Results]The main symptoms,secondary symptoms and total scores of TCM symptoms in the two groups were lower than those before treatment,and those in the observation group were lower after treatment(P<0.05).Compared with control group,the total clinical effective rate of observation group was higher(P<0.05),the time for body temperature returning to normal,the time for abdominal distension disappearing,the time for stool frequency and characteristics returning to normal were shorter in observation group after treatment(P<0.05).Compared with before treatment,GAS and MTL levels increased and SS levels decreased in the 2 groups after treatment(P<0.05).Compared with the control group,GAS and MTL levels were higher in the observation group,while SS levels were lower(P<0.05).There was no significant difference in total incidence of adverse reactions between the two groups(P>0.05).[Conclusion]Acupoint application combined with Xingpi Yang'er Granules in the treatment of children with dyspeptic diarrhea is effective,can obviously improve clinical symptoms,shorten the recovery process,adjust the level of gastrointestinal hormone,good safety.

Objective:To investigate the relationship between the expression levels of serum glycogen synthase kinase-3β (GSK-3β) and mothers against decapentaplegic homolog 4 (SMAD4) in patients with acute coronary syndrome (ACS) and the severity and prognosis of coronary artery disease.Methods:A total of 192 ACS patients admitted to Shandong First Medical University Affiliated Qingdao Hospital from June 2020 to May 2022 were selected as the ACS group, while 192 non ACS patients admitted to the same hospital were selected as the non ACS group. Enzyme-linked immunosorbent assay method was applied to determine the expression levels of serum GSK-3β and SMAD4 in two groups. The Gensini score was applied to evaluate the degree of coronary artery disease in patients, Spearman method was applied to analyze the correlation between serum GSK-3β, SMAD4 expression levels and Gensini score in ACS patients. Receiver operating characteristic (ROC) curve was applied to analyze the predictive efficacy of serum GSK-3β and SMAD4 levels on the prognosis of ACS patients.Results:The serum levels of GSK-3β and SMAD4 in the ACS group were significantly higher than those in the non ACS group: (2.70 ± 0.40) μg/L vs. (2.24 ± 0.41) μg/L, (12.19 ± 2.10) μg/L vs. (9.79 ± 2.82) μg/L, and there were statistical differences ( P<0.05). The serum levels of GSK-3β and SMAD4 in ACS patients with mild, moderate and severe coronary artery disease increased sequentially ( P<0.05). Spearman analysis showed that serum GSK-3β and SMAD4 levels in ACS patients were positively correlated with Gensini total score ( r = 0.569 and 0.587, P<0.01). In ACS patients, 48 cases had poor prognosis, and the incidence of poor prognosis was 25.00% (48/192); 144 cases had a good prognosis. The serum levels of GSK-3β and SMAD4 in ACS patients with poor prognosis were significantly higher than those in ACS patients with good prognosis: (3.15 ± 0.53) μg/L vs. (2.55 ± 0.36) μg/L, (14.03 ± 2.08) μg/L vs. (11.58 ± 2.11) μg/L, and there were statistical differences ( P<0.05). The ROC curve result indicated that the area under the curve (AUC) for predicting poor prognosis in ACS patients with serum GSK-3β and SMAD4 levels alone and in combination was 0.799, 0.784 and 0.858, respectively, the AUC predicted by the combination of the two was obviously higher than the AUC predicted separately ( Z = 2.04 and 2.75, P = 0.041 and 0.006). Conclusions:The expression levels of GSK-3β and SMAD4 in the serum of ACS patients are abnormally elevated. They are positively correlated with the degree of coronary artery disease in ACS patients, and both have good predictive power for adverse prognosis in ACS patients, while the combined use of the two has better predictive performance.

Objective To investigate the therapeutic efficacy of combining fractional CO2 laser with triam-cinolone acetonide and econazole nitrate cream for the treatment of vulvar lichen sclerosus(VLS).Methods The clinical data of 113 patients with vulvar lichen sclerosus(VLS)were retrospectively analyzed.The patients were divided into an observation group and a control group based on different treatments received.Clinical symptoms(vulvar itching,painful intercourse)and signs(vulvar skin color,vulvar skin elasticity,range of vulvar lesions)were scored using the Cattaneo scoring criteria.The Cattaneo scores before and after treatment were recorded for each patient,and comparisons were made between groups in terms of Cattaneo scores and occurrence of adverse reac-tions.The therapeutic efficacy of fractional CO2 laser combined with triamcinolone acetonide and econazole nitrate cream was evaluated.Results After three treatments,both groups exhibited a significant decrease in Cattaneo scores,with a statistically significant difference(P<0.05).The study group demonstrated a significantly greater reduction in Cattaneo scores compared to the control group(P<0.05).Furthermore,the cure rate of VSL was higher in the study group than in the control group(25.86%vs.10.91%,P<0.05).Regarding adverse effects,there was no statistically significant difference between the two groups(P>0.05).Conclusions Compared to the single application of fractional CO2 laser,the combination of fractional CO2 laser with triamcinolone acetonide and econazole nitrate cream demonstrates enhanced efficacy in alleviating signs and symptoms among patients with vulvar sclerosing tundra,while minimizing adverse reactions.This approach holds significant potential for clinical implementation.