In recent years, China has been facing the dual challenges of declining fertility rates and births, with male reproductive health issues, especially the decline in semen quality, identified as a pivotal contributor to this phenomenon. Meanwhile, accumulating evidence indicates that air pollutants, an increasingly severe environmental problem, can damage semen quality not only directly through their biological toxicity but also indirectly by disrupting the composition of microbial communities in the gut and semen, thereby dysregulating immune function, endocrine homeostasis, and oxidative stress responses. The gut microbiota and semen microbiota, as important components of the human microecosystem, play crucial roles in maintaining reproductive health. This article comprehensively reviewed the research progress on the potential effects of air pollutants (particulate matter and gaseous pollutants), gut microbiota, and semen microbiota on semen quality. Specifically, it elucidated the mechanisms of interaction between these factors and explored how they affect male fertility.

Definitive treatment of lung cancer with radiotherapy is challenging, as respiratory motion and anatomical changes can increase the risk of severe off-target effects during radiotherapy. Online adaptive radiotherapy (ART) is an evolving approach that enables timely modification of a treatment plan during the interfraction of radiotherapy, in response to physiologic or anatomic variations, aiming to improve the dose distribution for precise targeting and delivery in lung cancer patients. The effectiveness of online ART depends on the seamless integration of multiple components: sufficient quality of linear accelerator-integrated imaging guidance, deformable image registration, automatic recontouring, and efficient quality assurance and workflow. This review summarizes the present status of online ART for lung cancer, including key technologies, as well as the challenges and areas of active research in this field.
Humans ; Lung Neoplasms/radiotherapy* ; Radiotherapy Planning, Computer-Assisted/methods*

Microalgae possess high photosynthetic efficiency, robust adaptability, and substantial biomass, serving as excellent biological resources for large-scale cultivation. Malic enzyme (ME), a ubiquitous metabolic enzyme in living organisms, catalyzes the decarboxylation of malate to produce pyruvate, CO₂, and NAD(P)H, playing a role in multiple metabolic pathways including energy metabolism, photosynthesis, respiration, and biosynthesis. In this study, we identified the Scenedesmus quadricauda malic enzyme gene (SqME) and its biological functions, aiming to provide excellent target genes for the genetic improvement of higher plants. Based on the RNA-seq data from S. quadricauda under the biofilm cultivation mode with high CO₂ and light energy transfer efficiency and small water use, a highly expressed gene (SqME) functionally annotated as ME was cloned. The physicochemical properties of the SqME-encoded protein were systematically analyzed by bioinformatics tools. The subcellular localization of SqME was determined via transient transformation in Nicotiana benthamiana leaves. The biological functions of SqME were identified via genetic transformation in Nicotiana tabacum, and the potential of SqME in the genetic improvement of higher plants was evaluated. The ORF of SqME was 1 770 bp, encoding 590 amino acid residues, and the encoded protein was located in chloroplasts. SqME was a NADP-ME, with the typical structural characteristics of ME. The ME activity in the transgenic N. tabacum plant was 1.8 folds of that in the wild-type control. Heterologous expression of SqME increased the content of chlorophyll a, chlorophyll b, and total chlorophyll by 20.9%, 26.9%, and 25.2%, respectively, compared with the control. The transgenic tobacco leaves showed an increase of 54.0% in the fluorescence parameter NPQ and a decrease of 30.1% in Fo compared with the control. Moreover, the biomass, total lipids, and soluble sugars in the transgenic tobacco leaves enhanced by 20.5%, 25.7%, and 9.5%, respectively. On the contrary, the starch and protein content in the transgenic tobacco leaves decreased by 22.4% and 12.2%, respectively. Collectively, the SqME-encoded protein exhibited a strong enzymatic activity. Heterologous expressing of SqME could significantly enhance photosynthetic protection, photosynthesis, and biomass accumulation in the host. Additionally, SqME can facilitate carbon metabolism remodeling in the host, driving more carbon flux towards lipid synthesis. Therefore, SqME can be applied in the genetic improvement of higher plants for enhancing photosynthetic carbon fixation and lipid accumulation. These findings provide scientific references for mining of functional genes from S. quadricauda and application of these genes in the genetic engineering of higher plants.
Nicotiana/genetics* ; Photosynthesis/physiology* ; Malate Dehydrogenase/biosynthesis* ; Plant Leaves/genetics* ; Scenedesmus/enzymology* ; Carbon Cycle/genetics* ; Lipid Metabolism/genetics* ; Plants, Genetically Modified/metabolism*

Ovulatory disorders are mainly characterized by abnormal follicular maturation or ovulation， with complex etiologies and a lack of effective prevention and treatment methods. Autophagy dysfunction is closely related to the generation and progression of ovulatory disorders. This article systematically elucidates the mechanisms of TCM on follicular development and ovulatory disorders from the perspective of autophagy. It also reviews relevant studies on how TCM regulates autophagy to influence follicular development and improve ovulatory disorders. The findings reveal that TCM monomers/active ingredients （leonurine， total flavonoids from Eucommia ulmoides， alpinetin， icariin， etc.） and compound formulas （including Cangfu daotan decoction， Guishen yugong decoction， Zhuluan decoction， Yishen yangluan formula， Guishen pill， etc.） improve the follicular microenvironment， regulate sex hormone levels， and reduce follicular atresia by regulating autophagy-related genes and signaling pathways such as phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin and AMP-activated protein kinase. These actions thereby promote normal follicular development and ovulation， and delay ovarian aging. Most research in this field is based on cellular and animal experiments， often focusing on a single signaling pathway or factor. Some studies fail to fully reflect the individualized treatment characteristics of TCM that emphasize “syndrome differentiation and treatment”， highlighting the urgent need for further investigation.

Objective To analyze the clinicopathological characteristics and survival prognosis of young female patients with gastric adenocarcinoma. Methods A retrospective analysis was conducted on female patients who underwent radical gastrectomy at Zhongshan Hospital, Fudan University between January 2014 and December 2020, with postoperative pathological confirmation of gastric adenocarcinoma. Those aged ≤45 years were defined as the young group (n＝287), and were matched in a 1∶2 ratio by pTNM stage with female patients aged ≥60 years (elderly group, n＝574). Clinicopathological characteristics were compared between the two groups. Survival curves were plotted using the Kaplan-Meier method, and overall survival (OS) rates were assessed by log-rank test. Prognostic factors in the young group were analyzed using Cox regression models. Results Compared to elderly patients, young female gastric cancer patients exhibited a higher prevalence of tumors in the middle third of the stomach and a lower proportion in the upper third of the stomach. Molecular profiling revealed a higher frequency of HER2-low expression and elevated Ki-67 index. Pathologically, these patients were more frequently diagnosed with poorly differentiated or undifferentiated adenocarcinoma and signet ring cell carcinoma, while Lauren classification showed a predominance of the diffuse type with a lower proportion of the intestinal type (P＜0.05). Stratified analysis showed no significant difference in OS rates between the two groups for stage Ⅱ and Ⅲ patients; however, among stage Ⅰ patients, the young group had significantly better OS rates than the elderly group (P＝0.037). Multivariate Cox analysis and log-rank test confirmed that pN₃ stage (HR＝3.576, 95%CI 1.652–7.740), stage Ⅲ (HR＝3.581, 95%CI 1.059–12.106), and diffuse type (HR＝2.711, 95%CI 1.316–5.585) were risk factors for poor prognosis in young female gastric cancer patients. Conclusions Young female patients with gastric adenocarcinoma typically present with clinicopathological features such as the diffuse type, poor differentiation, and high proliferation. Moreover, pN₃ stage, stage Ⅲ cancer, and the diffuse type histology are correlated with a poor prognosis in this demographic.

BACKGROUND:Exercise training is an important non-drug rehabilitation method for many heart diseases,and it can also enhance the heart's tolerance to adverse stressors(such as myocardial ischemia),that is,exercise preconditioning.Granulocyte colony-stimulating factor can effectively mobilize stem cell homing and differentiation and promote the repair of damaged myocardium.However,the effect of the combination of the two treatments is not yet clear. OBJECTIVE:To explore the effect of granulocyte colony-stimulating factor supplementation combined with high-intensity intermittent exercise preconditioning on cardiac remodeling in rats with acute myocardial infarction and investigate its possible mechanism. METHODS:Totally 58 male Wistar rats were divided into sham group(n=10),model group(n=12),model drug group(n=12),model exercise group(n=12)and model combined treatment group(n=12).The myocardial infarction rat model was made by coronary artery ligation.The model exercise group and the model combined treatment group were pretreated with 8 weeks of high-intensity intermittent exercise on an electric treadmill before modeling.The model drug group and the model combined treatment group were subcutaneously injected with human recombinant granulocyte colony-stimulating factor 3 hours after modeling for 5 days(10 μg/kg per day).At 8 weeks after administration,echocardiography was used to detect heart structure and function;heart was stained with 2,3,5-triphenyltetrazolium chloride and Masson staining to obtain myocardial infarct area and collagen volume fraction,respectively.Vessel density and cell apoptosis rate were detected by immunofluorescence.Real-time fluorescent quantitative PCR was utilized to detect the mRNA expression of embryonic genes(brain natriuretic peptide,β-myosin heavy chain)and myocardial contraction regulatory factors(α-myosin heavy chain,sarcoplasmic reticulum Ca2+-ATPase).Western blot assay was used to detect the protein expression of cardiac stromal cell-derived factor 1,CXC chemokine receptor protein 4,Janus kinase 2(JAK2),signal transducer and activator of transcription 3(STAT3),Bcl2,Bax,and cleaved caspase-3. RESULTS AND CONCLUSION:(1)Compared with sham group,myocardial infarct size,collagen volume fraction,and apoptosis rate increased(P<0.05);vessel density,left ventricular fractional shortening,and left ventricular ejection fraction decreased(P<0.05);brain natriuretic peptide and β-myosin heavy chain mRNA increased(P<0.05),α-myosin heavy chain and sarcoplasmic reticulum Ca2+-ATPase mRNA and α-myosin heavy chain/β-myosin heavy chain ratio decreased(P<0.05);stromal cell-derived factor 1,CXC chemokine receptor protein 4,Bax,cleaved caspase-3 protein expression increased(P<0.05);p-JAK2,p-STAT3,and Bcl-2 protein expression decreased(P<0.05)in the model group.(2)Compared with the model group,the above indexes in the model drug and model exercise groups were significantly improved(P<0.05).Compared with the model drug and model exercise groups,the above parameters were further ameliorated(P<0.05)in the model combined treatment group.(3)The results showed that supplementation of granulocyte colony-stimulating factor or high-intensity intermittent exercise preconditioning alone can improve cardiac remodeling in rats with acute myocardial infarction,and the combined therapy has a better effect,which may be related to the induction of stem cell homing and the activation of JAK2/STAT3 signaling pathway to inhibit cardiomyocyte apoptosis.

OBJECTIVE To establish two-dimensional liquid chromatography method for the determination of imipenem blood concentration and apply it in clinical practice. METHODS The method for the determination of imipenem blood concentration was established based on automatic two-dimensional liquid chromatography. The targets were extracted by 1-dimensional column Aston SNCB （50 mm ×4.6 mm， 5 μm） and further separated and determined by 2-dimensional column Aston SCB （250 mm×4.6 mm， 5 μm）. The 1-dimensional mobile phase was imipenem-1D mobile phase [acetonitrile-methanol-water （15∶10∶75， V/V/V）] with a flow rate of 1.0 mL/min； 2-dimensional mobile phase was 72%OPI-1 organic mobile phase （chromatographic grade methanol）-20% BPI-1 alkaline mobile phase [water （containing 20.0 mmol/L ammonium phosphate， pH adjusted to 7.2 with triethylamine）]-8%API-1 acidic mobile phase [water （containing 20.0 mmol/L ammonium phosphate， pH adjusted to 3.0 with phosphoric acid）] with a flow rate of 1.0 mL/min； the column temperature was 40 ℃， UV detection wavelength was 310 nm and injection volume was 100 μL. Elution procedure： 1-dimensional column consisted of imipenem-1D mobile phase with eluting for 0-3.40 min； 2-dimensional column consisted of 72% OPI-1 organic mobile phase-20%BPI-1 alkaline mobile phase-8%API-1 acidic mobile phase with eluting for 3.40-11.00 min. RESULTS The linear range of imipenem was 0.171-18.570 μg/mL （R 2＝0.999 9） with the lower limit of quantification for 0.171 μg/mL； the recovery rate ranged from 93.47% to 106.16%（ n＝5） and the RSDs of both intra-day and inter- day precision were below 15% （n＝5）. The minimum concentration of imipenem in 51 patients ranged from 0 to 19.57 μg/mL. CONCLUSIONS The established method is simple and fast with the large scale of sample， and can be used for the imipenem blood concentration monitoring in patients with severe infection.

Objective:To investigate the risk factors for lymph node metastasis in patients with early gastric cancer and establish a model for prediction of risk.Methods:The cohort of this retrospective observational study comprised 1096 patients who had undergone radical gastric cancer surgery combined with standard D1 lymphadenectomy and been diagnosed with early gastric cancer by postoperative pathology in Zhongshan Hospital affiliated with Fudan University from January 2016 to July 2022. The patients were allocated to groups with and without lymph node metastases. Clinicopathological characteristics were compared between the two groups and multi-factor logistic regression analysis used to identify independent risk factors for lymph node metastasis in patients with early gastric cancer. Indications for endoscopic resection in the Japanese Gastric Cancer Association (JGCA) guideline were also incorporated into construction of the model. The patient cohort was divided into training and validation sets in a 6:4 ratio. The identified independent risk factors were used to construct a predictive nomogram. Receiver operating characteristic curves were plotted separately and the difference between them in predictive efficacy was compared using the area under the curve (AUC).Results:A total of 1,096 patients with early gastric cancer were included, with 750 males and 346 females. Their average age was (61.4±10.9) years old, and the mean tumor diameter was (23.8±11.4) mm. Among them, 188 patients (17.2%) had positive lymph node metastasis, with 109 cases in N1 stage, 42 cases in N2 stage, and 37 cases in N3 stage. Additionally, 462 patients were in T1a stage, while 634 patients were in T1b stage. Univariate analysis showed that tumor diameter, location, Lauren classification, gross morphology, histological type, intravascular invasion, ulceration, differentiation type and tumor T stage were associated with lymph node metastasis after radical gastrectomy for early gastric cancer (all P<0.05). Multifactorial analysis showed that the presence of intravascular invasion (OR=14.822, 95%CI: 9.323–23.572, P<0.001), undifferentiated type (OR=3.095, 95%CI: 1.649–5.811, P<0.001), tumor T1b (OR=1.798, 95%CI: 1.053–3.079, P=0.032), and tumor diameter ≥2 cm (OR=1.229, 95%CI: 1.031–1.469, P=0.022) were independent risk factors for lymph node metastasis. The baseline data of the training set and validation set were consistent in terms of balance (all P>0.05). We used the above variables to establish a predictive nomogram for lymph node metastasis in patients with early gastric cancer. The AUC values obtained from the validation of the model in the training and validation sets were 0.880 (95%CI: 0.849–0.911) and 0.881 (95%CI: 0.841–0.921), respectively, and were significantly better than the predictive efficacy based on the JGCA guideline (AUC=0.777, 95%CI: 0.746–0.809, P<0.001). Conclusions:Patients with early gastric cancer and intravascular invasion, undifferentiated tumors, tumor T1b, and diameter ≥2 cm are at higher risk of postoperative lymph node metastasis than other patients. The predictive model developed in this study more accurately predicts lymph node metastasis in patients with early gastric cancer than previously proposed methods.