Objective: To investigate the effects of "liver-smoothing and spirit-regulating" acupuncture on the intestinal flora, lipopolysaccharide (LPS) and the hippocampal toll-like receptor 4 (TLR4)/ transcription factor (NF)-κB signaling pathway in depressive disorder mouse model, and to explore its underlying mechanisms. Methods: Eighteen male SPF-grade C57BL/6J mice were randomly assigned to the control, model, and acupuncture groups using a random number method, with six mice in each group. The depression disorder model was induced in mice from both the acupuncture and model groups using CUMS. The mice in the acupuncture group were treated with acupuncture at the acupoints of "Baihui" (DU20), "Yintang" (DU29), "Hegu" (LI4), and "Taichong" (LR3) on the 15th day of modeling, with a duration of 20 min per session, once per day, for 2 consecutive weeks. Behavioral differences were assessed using the sucrose preference test, open field test, and forced swim test. Hematoxylin-eosin staining was used to observe pathological changes in the hippocampus and colon. The levels of the inflammatory factors interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and LPS in the hippocampus and colon were measured using Enzyme-linked Immunosorbent Assay. Western blotting was used to detect the expression of TLR4 and NF-κB protein in the hippocampus. Changes in gut microbiota structure and abundance were analyzed by 16 S rDNA sequencing. Results: Compared to the control group, the model group showed reduced sucrose preference rate, time in the center area, and total distance, with an increase in immobility time (P<0.01). Inflammatory pathological changes were observed in the hippocampal CA1 region and colon. The contents of IL-1β, IL-6, TNF-α, and LPS in the hippocampus and colon increased (P<0.01). The protein expression levels of hippocampal TLR4 and NF-κB were increased (P<0.01). The Chao1 index was increased (P<0.01). The relative abundances of Pseudomonadales, Acinetobacter, Moraxellaceae, Solibacillus, Escherichia_shigella, Enterobacteriaceae, Enterobacterales, Dubosiella, and Erysipelottichales were decreased, while the relative abundances of Alloprevotella and gram_negative_bacteriurh_cTPY_13 were increased (P<0.05). The pathways of lipopolysaccharide biosynthesis and pathogenic Escherichia coli infection were upregulated, and the pathway of terpenoid backbone biosynthesis was downregulated (P<0.01). Compared to the model group, the acupuncture group showed increased sucrose preference, time in the center area, and total distance, with a decrease in immobility time (P<0.01). The inflammatory pathological changes in the hippocampal CA1 region and colon were alleviated. The contents of IL-1β, IL-6, TNF-α, and LPS in the hippocampus and colon were reduced(P<0.01). The protein expression levels of hippocampal TLR4 and NF-κB were reduced (P<0.01). The Chao1 index was decreased (P<0.05), and the relative abundances of Dubosiella and Erysipelotrichaceae were increased, while the relative abundance of Rikenellaceae, Alloprevotella, and gram_negative_bacteriuch_cTPY_13 were decreased(P<0.05). The pathways of lipopolysaccharide biosynthesis and pathogenic Escherichia coli infection were significantly downregulated, and the pathway of terpenoid backbone biosynthesis was upregulated (P<0.01). Conclusion " Liver-smoothing and spirit-regulating" acupuncture can improve depressive symptoms in depressive disorder mice, potentially through regulating the LPS and TLR4/NF-κB signaling pathway mediated by intestinal flora, reducing the inflammatory response of the hippocampus, and improving the pathological injury of the hippocampus.

Objective To investigate the changes in the tumor microenvironment of pancreatic cancer(PDAC)complicated with diabetes mellitus(DM)in a mouse model of hyperglycemia and orthotopic pancreatic cancer by analyzing transcriptome and single-cell transcriptome data in order to identify potential therapeutic targets.Method By integrating single-cell transcriptome and bulk transcriptome data,bioinformatics analysis was conducted to compare the characteristics of tumor cells and tumor immune microenvironment between PDAC patients with DM(DM group)and those without DM(non-DM group).Twenty male C57BL/6 mice(6 weeks old,weighing 18～20 g)were randomly divided into a hyperglycemic group[STZ group,continuous intraperitoneal injection of 50 mg/kg streptozocin(STZ)(final concentration of 1％)dissolved in citrate buffer],and a control group(Control group,an equivalent volume of citrate buffer without STZ at the same time points),with 10 mice in each group.Tail-tip blood glucose level was measured to monitor glycemic status.After orthotopic inoculation of pancreatic cancer cells in both Control and STZ groups,tumor-infiltrating immune cells were harvested.Flow cytometry was employed to determine the effects of hyperglycemia on:total CD8+T cell and Treg cell populations;CD8+T cell subsets expressing Ki67,TNF-α,granzyme B(GZMB)and IFN-γ;surface expression of PD-1,lymphocyte activation gene-3(LAG-3)and T cell immunoglobulin and mucin domain-3(Tim-3)on CD8+T cells;programmed death-ligand 1(PD-L1)expression on tumor cells;and tumor-associated macrophage surface expression of major histocompatibility complex classⅠ(MHC-Ⅰ)and cluster of differentiation 206(CD206).Results Bioinformatics analysis revealed that,compared to the non-DM group,the genes significantly up-regulated in the DM group were associated with poor prognosis(P＜0.001).The proportion of type 2 ductal cells was increased in the DM group,exhibiting higher levels of copy number variation(P＜0.001).In the tumor immune microenvironment of the DM group,there was an increase in the proportion of Treg cells(P＜0.05)and an elevated exhaustion score for CD8+T cells(P＜0.001),accompanied by down-regulated expression of effector molecules,up-regulated expression of inhibitory checkpoints,and a significant increase in the M2 score of M2-like macrophages(P＜0.001).Animal experiments and flow cytometry found that,compared to the Control group,the STZ group had a shorter survival time(P＜0.001),with decreased proportions of total CD8+T cells(P＜0.01)and CD8+T cells expressing Ki67,TNF-α,GZMB and IFN-γ(P＜0.01),increased proportion of Treg cells(P＜0.001),up-regulated expression of PD-1,LAG-3 and Tim-3 on the surface of CD8+T cells(P＜0.001),and up-regulation of PD-L1 on tumor cell surface(P＜0.001)and enhanced expression of CD206 on the surface of tumor-associated macrophages,while down-regulated expression of MHC-Ⅰ(P＜0.001).Conclusion High glucose promotes the formation of an immunosuppressive microenvironment in PDAC,and targeting type 2 ductal cells and immunosuppressive cells in the tumor microenvironment,combined with dual immune checkpoint antibody therapy,may improve patient prognosis.

Objective To explore the role of mitophagy in pancreatic cancer cachexia-induced muscle atrophy and its underlying mechanism.Methods Six male C57BL/6J mice(8 weeks old,weighing 20～30 g)were equally and randomly divided into a control group(intrapancreatic injection of normal saline)and a cachexia group(orthotopic pancreatic injection of KPC1199 cells).After successful model establishment,gastrocnemius muscles were harvested for transmission electron microscopy(TEM)to assess mitochondrial ultrastructure.Western blotting was performed to quantify mitochondrial respiratory chain complexes(Ⅰ～Ⅳ)and autophagy-related proteins,while immunofluorescence staining was conducted to evaluate mitochondrial-lysosomal colocalization.In in vitro experiments,C2C12 myoblasts were differentiated into myotubes,and then divided into a control group(standard culture)and a cachexia group(co-cultured with KPC1199 cells for 48 h using transwell chambers).Mitochondrial-lysosomal colocalization and autophagy-related protein expression were analyzed with immunofluorescence assay and Western blotting.The mitochondrial division inhibitor Mdivi-1(20 μmol/L)was added to the co-culture system to assess its myotube diameter.Results Compared to the control mice,the cachectic mice exhibited mitochondrial swelling,reduced cristae density,and significantly increased mitochondrial-lysosomal colocalization in gastrocnemius muscle(P<0.05).Western blotting revealed the expression levels of mitochondrial respiratory chain proteins complexⅠ(1.00±0.04 vs 0.51±0.04,P<0.05),complexⅡ(1.00±0.13 vs 0.73±0.15,P<0.05),complexⅢ(1.00±0.20 vs 0.64±0.01,P<0.05),complexⅣ(1.00±0.06 vs 0.65±0.02,P<0.05)and PGC1α(1.00±0.03 vs 0.62±0.06,P<0.05)were decreased,and the levels of mitophagy markers,LC3-Ⅰ/Ⅱ(1.00±0.14 vs 1.65±0.25,P<0.05),PINK1(1.00±0.11 vs 1.51±0.05,P<0.05),and BNIP3(1.00±0.22 vs 2.02±0.10,P<0.05)were elevated when compared to the control.In the C2C12 myotube model,tumor cell co-culture increased mitochondrial-lysosomal colocalization and upregulated mitophagy-related protein expression(P<0.05),consistent with the in vivo findings.Mdivi-1 treatment increased myotube diameter from 220.6±35.5 μm to 315.0±39.1 μm(R2=0.666 5,P<0.05).Conclusion Mitophagy is activated in pancreatic cancer cachexia-induced muscle atrophy.Inhibiting mitophagy can effectively alleviate muscle atrophy induced by pancreatic cancer cachexia.

Objective:To summarize the prenatal ultrasound characteristics of fetal malformations with amniotic band syndrome (ABS) in early pregnancy, in order to improve the ultrasound detection rate of this disease.Methods:Pregnant women who underwent systematic ultrasound screening of early pregnancy fetal malformations in Beijing Obstetrics and Gynecology Hospital from January 2017 to October 2023 were retrospectively collected. Twenty-three cases of early pregnancy ABS were collected and confirmed by ultrasound diagnosis and induced abortion results. Clinical data, images, and examination results of these cases were studied and the ultrasound characteristics of early pregnancy ABS were summarized.Results:In all 23 cases, adhesions between the intrauterine amniotic band and the affected parts of the fetus were found during ultrasonography, and the malformations in each case were different and closely related to the damage to different body surface parts of the fetus. Multiple fetal malformations accounted for 78% (18/23) and single cases accounted for only 22% (5/23). Among the 23 cases, 19 (83%) involved fetal limbs, 11 (48%) brains, 11 (48%) thoracic and abdominal walls, 10 (43%) spine, 5 (22%) faces, and 5 (22%) umbilical cords.23 (100%) patients were confirmed to have ABS after induction of labor, which was consistent with the ultrasound diagnosis, but 11 (48%) of them did not accurately describe fetal limb or facial abnormalities.Conclusions:The fetal malformation of ABS in the first trimester is closely related to the amniotic band, which has the characteristics of multiple properties, uniqueness, randomness and asymmetry. The most affected parts are the fetal limbs, brain, and body wall, and severe malformations are common.Prenatal ultrasound can provide suggestive diagnosis.

Although somatic cells can be reprogrammed to pluripotent stem cells (PSCs) with pure chemicals, authentic pluripotency of chemically induced pluripotent stem cells (CiPSCs) has never been achieved through tetraploid complementation assay. Spontaneous reprogramming of spermatogonial stem cells (SSCs) was another non-transgenic way to obtain PSCs, but this process lacks mechanistic explanation. Here, we reconstructed the trajectory of mouse SSC reprogramming and developed a five-chemical combination, boosting the reprogramming efficiency by nearly 80- to 100-folds. More importantly, chemical induced germline-derived PSCs (5C-gPSCs), but not gPSCs and chemical induced pluripotent stem cells, had authentic pluripotency, as determined by tetraploid complementation. Mechanistically, SSCs traversed through an inverted pathway of in vivo germ cell development, exhibiting the expression signatures and DNA methylation dynamics from spermatogonia to primordial germ cells and further to epiblasts. Besides, SSC-specific imprinting control regions switched from biallelic methylated states to monoallelic methylated states by imprinting demethylation and then re-methylation on one of the two alleles in 5C-gPSCs, which was apparently distinct with the imprinting reprogramming in vivo as DNA methylation simultaneously occurred on both alleles. Our work sheds light on the unique regulatory network underpinning SSC reprogramming, providing insights to understand generic mechanisms for cell-fate decision and epigenetic-related disorders in regenerative medicine.
Male ; Mice ; Animals ; Cellular Reprogramming/genetics* ; Tetraploidy ; Pluripotent Stem Cells/metabolism* ; Induced Pluripotent Stem Cells/metabolism* ; DNA Methylation ; Spermatogonia/metabolism* ; Germ Cells/metabolism*

The anterior cruciate ligament (ACL) injury is a common sports injury that has a significant impact on knee function and patients′ mobility. With the popularity of national fitness campaign in China, the incidence of ACL injury is increasing year by year. Currently, there still lacks clinical standards or guidelines on how to choose appropriate treatment methods, surgical plans and rehabilitation protocols for ACL injury. In order to timely reflect the new treatment concept of ACL injury, standardize its diagnosis and treatment and improve the curative effect, the Sports Medicine Society of Chinese Research Hospital Association and the Editorial Board of Chinese Journal of Trauma organized domestic orthopedic and sports medicine experts to formulate the "clinical evidence-based guideline for the diagnosis and treatment of anterior cruciate ligament injury (2022 version)" based on the level of evidence-based medicine and in compliance with the principle of scientificity, practicability and advancement. The present guideline includes 12 recommendations for the diagnosis, treatment and rehabilitation of ACL injury in order to provide guidance and assistance for the clinical diagnosis and treatment of ACL injury in China.

Objective:To investigate and establish the related factors of non-invasive score model for prediction of non-alcoholic fatty liver disease in chronic hepatitis B patients with normal or mildly elevated alanine aminotransferase (ALT).Methods:A total of 128 cases with chronic hepatitis B who had undergone liver biopsy were included. According to the presence or absence of hepatocyte steatosis on the pathological results of liver biopsy, they were divided into a fatty infiltration and a non-fatty infiltration group. Patients' demographic characteristics, laboratory test indexes, and pathological test results were collected. Univariate and multivariate logistic regression analysis combined with clinical screening variables were used to establish a predictive model. The prediction efficiency of the new model was evaluated by the receiver operating curve, and the difference between the accuracy of the new model and ultrasound in the diagnosis of fatty liver was compared by Delong's-test.Result:Multivariate regression analysis showed that serum triglyceride, serum uric acid and platelets were highly correlated with intrahepatic steatosis ( P<0.05). The regression equation triglyceride-uric acid-platelet (TUP)-1=-8.195+0.011×uric acid+1.439×triglyceride+0.012×platelet count was established by combining the above variables. Tthe equation TUP-2=-7.527+0.010×uric acid+1.309×triglyceride+0.012×platelet count+1.397×fatty liver (ultrasound) was established (yes=1; no=0) after incorporating the results of abdominal ultrasound. The diagnostic value of TUP-1 and TUP-2 models for fatty liver was better than that of ultrasound alone and there was no statistically significant difference in diagnostic value between TUP-1 and TUP-2 models ( Z=1.453, P=0.146). Conclusion:Compared with abdominal ultrasonography alone, the new model is more effective in diagnosing fatty liver and has good application value.

ObjectiveTo investigate the efficacy and safety of venlafaxine in the treatment of depression under the guidance of pharmacogenomics testing， and to provide references for individualized medication. MethodsA total of 66 patients who met the diagnostic criteria of International Classification of Diseases， tenth edition （ICD-10） for depressive episode were included in the study. Patients who were recommended to be treated with venlafaxine in the pharmacogenomics testing report were divided into study group （n=32）， and those who were decided to be treated with venlafaxine by doctors after consultation with patients were divided into control group （n=34）. At the baseline and the end of the 2^nd， 4^th， 6^th and 8^th weekend of treatment， Hamilton Depression Scale-24 item （HAMD-24） was adopted to evaluate the clinical efficacy. Meanwhile， Sheehan Disability Scale （SDS） was applied to measure the social function of patients at the baseline and the end of the 8^th weekend of treatment. After treatment， Treatment Emergent Symptom Scale （TESS） was used to assess the incidence of adverse reactions. ResultsAt the end of the 4^th， 6^th and 8^th weekend of treatment， HAMD-24 scores in the study group were all lower than those in the control group， with statistical differences （t=2.344， 4.316， 5.760， P<0.05 or 0.01）. At the end of the 8^th weekend of treatment， SDS score of the study group was lower than that of the control group， with statistical difference （t=2.173， P<0.05）. The adverse reaction rate in the study group was lower than that in the control group， with statistical difference （χ²=5.720， P<0.05）. ConclusionTreatment of depression with venlafaxine based on pharmacogenetic testing is an effective and safe way to alleviate the depression symptoms in patients.

Objective:To investigate the incidence and risk factors of active human cytomegalovirus (HCMV) infection in patients with severe community-acquired pneumonia.Methods:Patients who required respiratory support and were diagnosed with severe community-acquired pneumonia in the respiratory intensive care unit (RICU) of the First Affiliated Hospital of Guangzhou Medical University from March 1, 2019 to June 1, 2020 were consecutively screened and divided into active HCMV infection group (20 cases) and non-active HCMV infection group (95 cases) based on whether a patient has active HCMV infection or not. Differences in demographic data, laboratory findings, and clinical outcomes were compared between the two groups. Moreover, logistic regression was applied to analyze risk factors for active HCMV infection.Results:The 20 of 115 patients with severe community-acquired pneumonia requiring respiratory support were confirmed to have active infection with HCMV, with a prevalence of active HCMV infection of 17.4%. The pneumonia severity index (PSI) and suppressor T lymphocytes (Ts) in active HCMV infection group were higher than that of the control group, and all the differences were statistically significant ( Z=2.432, P=0.015; Z=2.036, P=0.042); whereas lymphocytes, monocytes, blood lactate, and platelet levels were lower than those of the control group, and all the differences were statistically significant ( P < 0.05). Patients with active HCMV infection had a higher transfusion rate than the control group, and the differences were statistically significant (χ 2=3.941; P=0.047). Increasing levels of PSI and Ts percentage were independent risk factors for active HCMV infection ( OR=1.03, 95% CI: 1.01~1.05; OR=1.06, 95% CI: 1.00~1.11; P < 0.05). RICU length of stay, complication rates, and 90-day all-cause mortality were higher in the active HCMV infection group than the control group, and all the differences were statistically significant ( P < 0.05). Conclusions:Active HCMV infection is highly prevalent in patients with severe community-acquired pneumonia and associated with several adverse clinical outcomes, with PSI and Ts cell levels being independent risk factors.

Lumbar disc herniation (LDH) is one of the most common degenerative spinal diseases in adults and less common in adolescents. The age of onset is less than 21 years old and is defined as adolescent lumbar disc herniation (ALDH). The incidence of ALDH is only 1%-5%. However, with the accelerated pace of life, the incidence of ALDH is increasing. In recent years, the rapid development of endoscopic technology has been widely used in spinal surgery. ALDH's percutaneous endoscopic technique has achieved good results. Compared with open surgery, percutaneous endoscopic treatment can preserve more normal lumbar structures, such as ligaments, muscles, lamina and facets, minimizing the impact on pubertal growth. This article reviews the characteristics of ALDH and endoscopic treatment as an entry point.