1.Efficacy and mechanism of Guizhi Tongluo Tablets in alleviating atherosclerosis by inhibiting CD72hi macrophages.
Xing-Ling HE ; Si-Jing LI ; Zi-Ru LI ; Dong-Hua LIU ; Xiao-Jiao ZHANG ; Huan HE ; Xiao-Ming DONG ; Wen-Jie LONG ; Wei-Wei ZHANG ; Hui-Li LIAO ; Lu LU ; Zhong-Qi YANG ; Shi-Hao NI
China Journal of Chinese Materia Medica 2025;50(5):1298-1309
This study investigates the effect and underlying mechanism of Guizhi Tongluo Tablets(GZTL) in treating atherosclerosis(AS) in a mouse model. Apolipoprotein E-knockout(ApoE~(-/-)) mice were randomly assigned to the following groups: model, high-, medium-, and low-dose GZTL, and atorvastatin(ATV), and age-matched C57BL/6J mice were selected as the control group. ApoE~(-/-) mice in other groups except the control group were fed with a high-fat diet for the modeling of AS and administrated with corresponding drugs via gavage for 8 weeks. General conditions, signs of blood stasis, and body mass of mice were monitored. Aortic plaques and their stability were assessed by hematoxylin-eosin, Masson, and oil red O staining. Serum levels of total cholesterol(TC), triglycerides(TG), and low-density lipoprotein cholesterol(LDL-C) were measured by biochemical assays, and those of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6) were determined via enzyme-linked immunosorbent assay. Apoptosis was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL). Single-cell RNA sequencing(scRNA-seq) was employed to analyze the differential expression of CD72hi macrophages(CD72hi-Mφ) in the aortas of AS patients and mice. The immunofluorescence assay was employed to visualize CD72hi-Mφ expression in mouse aortic plaques, and real-time fluorescence quantitative PCR was utilized to determine the mRNA levels of IL-1β, TNF-α, and IL-6 in the aorta. The results demonstrated that compared with the control group, the model group exhibited significant increases in body mass, aortic plaque area proportion, necrotic core area proportion, and lipid deposition, a notable decrease in collagen fiber content, and an increase in apoptosis. Additionally, the model group showcased elevated serum levels of TC, TG, LDL-C, IL-1β, TNF-α, and IL-6, alongside marked upregulations in the mRNA levels of IL-1β, TNF-α, and IL-6 in the aorta. In comparison with the model group, the GZTL groups and the ATV group showed a reduction in body mass, and the medium-and high-dose GZTL groups and the ATV group demonstrated reductions in aortic plaque area proportion, necrotic core area proportion, and lipid deposition, an increase in collagen fiber content, and a decrease in apoptosis. Furthermore, the treatment goups showcased lowered serum levels of TC, TG, LDL-C, IL-1β, TNF-α, and IL-6. The data of scRNA-seq revealed significantly elevated CD72hi-Mφ signaling in carotid plaques of AS patients compared with that in the normal arterial tissue. Animal experiments confirmed that CD72hi-Mφ expression, along with several pro-inflammatory cytokines, was significantly upregulated in the aortas of AS mice, which were downregulated by GZTL treatment. In conclusion, GZTL may alleviate AS by inhibiting CD72hi-Mφ activity.
Animals
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Drugs, Chinese Herbal/administration & dosage*
;
Atherosclerosis/immunology*
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Mice
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Mice, Inbred C57BL
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Macrophages/immunology*
;
Male
;
Humans
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Apolipoproteins E/genetics*
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Tablets
;
Tumor Necrosis Factor-alpha/genetics*
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Apoptosis/drug effects*
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Interleukin-1beta/genetics*
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Interleukin-6/genetics*
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Disease Models, Animal
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Mice, Knockout
2.Postoperative Complications and 30-day Readmission in Patients Older than 80 Years with Chronic Kidney Disease after Hip Fracture.
Hua-Wen ZHANG ; Lu-Lu MA ; Xue-Rong YU
Chinese Medical Sciences Journal 2025;40(3):188-196
OBJECTIVES:
This study aimed to explore the impact of chronic kidney disease (CKD) on prognosis of patients older than 80 years after hip fracture.
METHODS:
This retrospective, observational, single-center study included patients older than 80 years who underwent hip fracture operations between Feburary 2013 to June 2021 at our hospital. Patients were divided into CKD and non-GKD groups based on the estimated glomerular filtration rate (eGFR) < 60 mL/(min·1.73m2)] or not. Outcomes were the incidence of in-hospital postoperative infectious and non-infectious complications, 30-day readmission, and in-hospital death. Logistic regression analysis was used to calculate the odds ratio (OR) of CKD on these outcomes.
RESULTS:
A total of 498 patients were included, 165 in the CKD group and 333 in the non-CKD group. Eighty-seven (52.7%) CKD patients experienced 140 episodes of postoperative complications. In comparison, 114 (34.2%) non-CKD patients had 158 episodes of postoperative complications. CKD patients were more likely to have postoperative complications than non-CKD patients (OR = 2.143, 95% CI: 1.465-3.134, P < 0.001). CKD increased the risk of cardiovascular complications (OR = 2.044, 95% CI: 1.245-3.356, P = 0.004), acute kidney injury (OR = 3.401, 95% CI: 1.905-6.072, P < 0.001), delirium (OR = 2.276, 95% CI: 1.140-4.543, P = 0.024), and gastrointestinal bleeding (OR = 4.151, 95% CI: 1.025-16.812, P = 0.031). The transfusion rate (OR = 2.457, 95% CI: 1.668-3.618, P < 0.001) and incidence of 30-day readmission (OR = 2.426, 95% CI:1.203-4.892, P = 0.011) in CKD patients were significantly higher than those in patients without CKD.
CONCLUSIONS
CKD is associated with poor postoperative outcomes in geriatric hip fracture patients. Special attention should be paid to patients with CKD.
Humans
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Renal Insufficiency, Chronic/physiopathology*
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Aged, 80 and over
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Postoperative Complications/epidemiology*
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Hip Fractures/complications*
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Male
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Female
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Patient Readmission/statistics & numerical data*
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Retrospective Studies
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Glomerular Filtration Rate
3.Clinical study on the effectiveness of bone acupuncture for alleviating pain and improving function in patients with degenerative lumbar spinal stenosis.
Chang-Xiao HAN ; Min-Shan FENG ; Jing-Hua GAO ; Xun-Lu YIN ; Guang-Wei LIU ; Hai-Bao WEN ; Jing LI ; Bo-Chen PENG ; Li-Guo ZHU
China Journal of Orthopaedics and Traumatology 2025;38(2):152-156
OBJECTIVE:
To assess the effectiveness of bone acupuncture in improving pain and function in degenerative lumbar spinal stenosis (DLSS) and compare it with Jiaji acupuncture.
METHODS:
From January to December 2023, 80 DLSS patients were treated with acupuncture and divided into bone acupuncture and Jiaji acupuncture groups. Among them, 40 patients in the bone acupuncture group included 15 males and 25 females, with a mean age of (60.60±6.98) years old;anthor 40 patients in the Jiaji acupuncture group included 16 males and 24 females, with a mean age of (61.48±9.55) years old. The Roland Morris disability questionnaire(RMDQ), walking distance, visual analogue scale(VAS), and the MOS item short from health survey(SF-36) of two groups at baseline, 2 weeks, 4 weeks, and 12 weeks post-treatment were compared.
RESULTS:
Eighty patients were followed up for 3 to 5 months with an average of (3.62±0.59) months. There was no significant differences in general data and the scores before treatment between two groups(P>0.05). The RMDQ scores in both groups decreased significantly at 2, 4 and 12 weeks after treatment compared with before treatment(P<0.05), at each time point after treatment, the decrease was more significant in the bone acupuncture group than in the Jiaji acupuncture group(P<0.05). The VAS of waist and leg in both groups was significantly lower at 2, 4 and 12 weeks after treatment that before treatment(P<0.05). At all time points after treatment, the waist VAS in the bone acupuncture group was reduced more significant than in the Jiaji acupuncture group(P<0.05);there was no significant difference in leg VAS at 2 and 12 weeks after treatment between two groups(P>0.05), the improvement was more significant in the bone acupuncture group in the 4 weeks after treatment than in the Jiaji acupuncture group. The SF-36 scores in both groups were significantly higher at 2, 4, and 12 weeks after treatment than before treatment(P<0.05);the SF-36 score raised more significant in the bone acupuncture group than in the Jiaji acupunture group(P<0.05). No significant difference in the walking distance between two groups at 2 weeks after treatment(P>0.05);the walking distance in the bone acupuncture group was significantly higher than that in the Jiaji acupuncture group at 4 and 12 weeks after treatment(P<0.05).
CONCLUSION
Bone-penetrating acupuncture moderately improves functional impairment, pain, and quality of life in patients with DLSS, showing better efficacy than Jiaji acupuncture.
Humans
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Female
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Male
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Middle Aged
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Acupuncture Therapy/methods*
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Spinal Stenosis/physiopathology*
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Aged
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Lumbar Vertebrae/physiopathology*
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Pain Management
4.Exploration of evaluation criteria based on the biological variation in the external quality assessment for basic semen analysis in China.
Xi-Yan WU ; Jin-Chun LU ; Xin-Hua PENG ; Jing-Liang HE ; Dao WANG ; Cong-Ling DAI ; Wen-Bing ZHU ; Gang LIU ; Wei-Na LI
Asian Journal of Andrology 2025;27(5):621-626
This study explores whether the current external quality assessment (EQA) level and acceptable bias for basic semen analysis in China are clinically useful. We collected data of semen EQA from Andrology laboratories in the Hunan Province (China) in 2022 and searched for data in the published literature from January 2000 to December 2023 in China. On the basis of these data, we analyzed the coefficients of variation and acceptable biases of different quality control materials for basic semen analysis through robust statistics. We compared these findings with quality specifications based on biological variation from optimal, desirable, and minimum levels of bias to seek a unified and more suitable semen EQA bias evaluation standard for China's national conditions. Different sources of semen quality control material exhibited considerable variation in acceptable biases among laboratories, ranging from 8.2% to 56.9%. A total of 50.0% of the laboratories met the minimum quality specifications for progressive motility (PR), whereas 100.0% and 75.0% of laboratories met only the minimum quality specifications for sperm concentration and total motility (nonprogressive [NP] + PR), respectively. The Z value for sperm concentration and PR+NP was equivalent to the desirable performance specification, whereas the Z value for PR was equivalent only to the minimum performance specification. This study highlights the feasibility of operating external quality assessment schemes for basic semen analysis using quality specifications based on biological variation. These specifications should be unified among external quality control (EQC) centers based on biological variation.
Semen Analysis/standards*
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Humans
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China
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Male
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Quality Control
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Sperm Motility
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Sperm Count/standards*
5.Comparison of the therapeutic efficacy of different methods of anesthesia in microscopic varicocelectomy for the treatment of varicocele.
Qun-Sheng LI ; Ning-Hua LI ; Lei ZHOU ; Dong-Run LI ; Jie LU ; Chun-Yan HE ; Yu-Nu ZHOU ; Jian-Mo CHEN ; Wen-Tao YANG
National Journal of Andrology 2025;31(8):692-697
OBJECTIVE:
To compare the therapeutic efficacy and safety of local anesthesia and spinal anesthesia for the patients with varicocele (VC) who underwent microsurgical varicocelectomy (MV).
METHODS:
We retrospectively analyzed the data of VC patients who underwent MV treatment at the Andrology Department of the Affiliated Ruikang Hospital of Guangxi University of Chinese Medicine from May 2020 to March 2023. Cases with complete clinical data and follow-up evaluation were selected and divided into a control group (spinal anesthesia) and an observation group (local anesthesia) according to different anesthesia methods. The surgical time (including anesthesia time), visual analogue scale (VAS) score for pain, hospital stay, treatment cost, sperm concentration, forward motile sperm rate, and normal sperm morphology rate after three months of surgery, as well as postoperative complications and recurrence rate were compared between the two groups.
RESULTS:
A total of 107 eligible cases were included, with 56 cases in the control group and 51 cases in the observation group. There was no significant difference in the VAS score for pain during and after four hours of surgery, as well as postoperative complications, and recurrence rate between the two groups (P> 0.05). There was an significant increase in sperm concentration, forward motile sperm rate, and normal sperm morphology rate in both of two groups after three months of surgery (P<0.05). However, there was no significant difference between the two groups three months after surgery (P>0.05). The surgical time and hospital stay were shorter than those of the control group (P<0.05). And the treatment cost in observation group was lower than that of the control group (P<0.05).
CONCLUSION
Both local anesthesia and lumbar anesthesia for MV treatment of VC have good efficacy and safety. However, patients treated with MV under local anesthesia for VC have obvious advantages in terms of operation time (including anesthesia time), hospital stay, and treatment cost, which is worthy of clinical promotion and application.
Humans
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Male
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Varicocele/surgery*
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Retrospective Studies
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Microsurgery
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Anesthesia, Spinal
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Adult
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Treatment Outcome
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Anesthesia, Local
6.Psychological stress-activated NR3C1/NUPR1 axis promotes ovarian tumor metastasis.
Bin LIU ; Wen-Zhe DENG ; Wen-Hua HU ; Rong-Xi LU ; Qing-Yu ZHANG ; Chen-Feng GAO ; Xiao-Jie HUANG ; Wei-Guo LIAO ; Jin GAO ; Yang LIU ; Hiroshi KURIHARA ; Yi-Fang LI ; Xu-Hui ZHANG ; Yan-Ping WU ; Lei LIANG ; Rong-Rong HE
Acta Pharmaceutica Sinica B 2025;15(6):3149-3162
Ovarian tumor (OT) is the most lethal form of gynecologic malignancy, with minimal improvements in patient outcomes over the past several decades. Metastasis is the leading cause of ovarian cancer-related deaths, yet the underlying mechanisms remain poorly understood. Psychological stress is known to activate the glucocorticoid receptor (NR3C1), a factor associated with poor prognosis in OT patients. However, the precise mechanisms linking NR3C1 signaling and metastasis have yet to be fully elucidated. In this study, we demonstrate that chronic restraint stress accelerates epithelial-mesenchymal transition (EMT) and metastasis in OT through an NR3C1-dependent mechanism involving nuclear protein 1 (NUPR1). Mechanistically, NR3C1 directly regulates the transcription of NUPR1, which in turn increases the expression of snail family transcriptional repressor 2 (SNAI2), a key driver of EMT. Clinically, elevated NR3C1 positively correlates with NUPR1 expression in OT patients, and both are positively associated with poorer prognosis. Overall, our study identified the NR3C1/NUPR1 axis as a critical regulatory pathway in psychological stress-induced OT metastasis, suggesting a potential therapeutic target for intervention in OT metastasis.
7.A Retrospective Study of Pregnancy and Fetal Outcomes in Mothers with Hepatitis C Viremia.
Wen DENG ; Zi Yu ZHANG ; Xin Xin LI ; Ya Qin ZHANG ; Wei Hua CAO ; Shi Yu WANG ; Xin WEI ; Zi Xuan GAO ; Shuo Jie WANG ; Lin Mei YAO ; Lu ZHANG ; Hong Xiao HAO ; Xiao Xue CHEN ; Yuan Jiao GAO ; Wei YI ; Yao XIE ; Ming Hui LI
Biomedical and Environmental Sciences 2025;38(7):829-839
OBJECTIVE:
To investigate chronic hepatitis C virus (HCV) infection's effect on gestational liver function, pregnancy and delivery complications, and neonatal development.
METHODS:
A total of 157 HCV antibody-positive (anti-HCV[+]) and HCV RNA(+) patients (Group C) and 121 anti-HCV(+) and HCV RNA(-) patients (Group B) were included as study participants, while 142 anti-HCV(-) and HCV RNA(-) patients (Group A) were the control group. Data on biochemical indices during pregnancy, pregnancy complications, delivery-related information, and neonatal complications were also collected.
RESULTS:
Elevated alanine aminotransferase (ALT) rates in Group C during early, middle, and late pregnancy were 59.87%, 43.95%, and 42.04%, respectively-significantly higher than Groups B (26.45%, 15.70%, 10.74%) and A (23.94%, 19.01%, 6.34%) ( P < 0.05). Median ALT levels in Group C were significantly higher than in Groups A and B at all pregnancy stages ( P < 0.05). No significant differences were found in neonatal malformation rates across groups ( P > 0.05). However, neonatal jaundice incidence was significantly greater in Group C (75.16%) compared to Groups A (42.25%) and B (57.02%) ( χ 2 = 33.552, P < 0.001). HCV RNA positivity during pregnancy was an independent risk factor for neonatal jaundice ( OR = 2.111, 95% CI 1.242-3.588, P = 0.006).
CONCLUSIONS
Chronic HCV infection can affect the liver function of pregnant women, but does not increase the pregnancy or delivery complication risks. HCV RNA(+) is an independent risk factor for neonatal jaundice.
Humans
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Female
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Pregnancy
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Adult
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Pregnancy Complications, Infectious/epidemiology*
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Retrospective Studies
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Pregnancy Outcome
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Infant, Newborn
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Viremia/virology*
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Hepatitis C
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Hepacivirus/physiology*
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Hepatitis C, Chronic/virology*
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Young Adult
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Alanine Transaminase/blood*
8.Study on the clinical relationship between inflammatory burden index and gastric cancer
Lin YANG ; Ning LU ; Hua WEN ; Mingxin ZHANG ; Lin ZHU
Journal of International Oncology 2024;51(5):274-279
Objective:To analyze the relationship between inflammatory burden index (IBI) and the diagnosis and prognosis of gastric cancer.Methods:A total of 180 patients with gastric cancer admitted to the First Affiliated Hospital of Xi'an Medical University from January 1, 2017 to December 31, 2021 were selected as the gastric cancer group, 180 cases of gastric precancerous diseases treated in this hospital during the same period were selected as the precancerous disease group, 180 healthy individuals who underwent physical examinations in this hospital during the same period were selected as the control group. The value of IBI in the diagnosis of gastric cancer was analyzed. The IBI values of the three groups were compared by one-way analysis of variance, and the optimal cut-off values of different indicators were determined by receiver operator characteristic (ROC) curve to further evaluate the diagnostic value of IBI for gastric cancer. LASSO regression analysis was performed using the R software package glmnet to screen variables. Univariate and multivariate Cox regression analysis were performed on the screened variables to further determine the independent risk factors affecting the prognosis of patients with gastric cancer.Results:The levels of IBI in gastric cancer, precancerous disease, and control group were 46.69±8.38, 35.53±6.11, and 21.47±4.36, respectively, with a statistically significant difference ( F=681.45, P<0.001). Further pairwise comparisons showed that the gastric cancer group was higher than the precancerous disease group and the control group, and the precancerous disease group was higher than the control group, with statistically significant differences ( t=14.43, P<0.001; t=34.15, P<0.001; t=25.13, P<0.001). ROC curve analysis showed that the optimal cut-off value for the neutrophil to lymphocyte ratio (NLR) was 3.15 (AUC=0.65, 95% CI: 0.54-0.73), with a specificity of 92.31% and a sensitivity of 43.97%. The optimal cut-off value for the platelet to lymphocyte ratio (PLR) was 137.36 (AUC=0.59, 95% CI: 0.51-0.68), with a specificity of 93.66% and a sensitivity of 34.98%. The optimal cut-off value for lymphocyte to monocyte ratio (LMR) was 5.18 (AUC=0.69, 95% CI: 0.58-0.76), with a specificity of 61.29% and a sensitivity of 73.33%. The optimal cut-off value for panimmune inflammation value (PIV) was 276.57 (AUC=0.73, 95% CI: 0.65-0.78), with a specificity of 78.41% and a sensitivity of 59.77%. The optimal cut-off value for the systemic immune inflammatory index (SII) was 654.37 (AUC=0.73, 95% CI: 0.66-0.78), with a specificity of 86.32% and a sensitivity of 62.11%. The optimal cut-off value for the IBI was 49.12 (AUC=0.82, 95% CI: 0.79-0.92), with a specificity of 85.36% and a sensitivity of 65.82%. LASSO regression analysis showed that there were 8 prognostic variables related to survival, including IBI, T stage, N stage, age, carcinoembryonic antigen (CEA), SII, PIV, and tumor grade. The results of univariate analysis showed that IBI ( HR=4.57, 95% CI: 3.82-5.32, P<0.001), T stage ( HR=2.54, 95% CI: 1.75-3.31, P<0.001), N stage ( HR=1.69, 95% CI: 1.15-2.21, P=0.032), CEA ( HR=1.38, 95% CI: 1.06-1.75, P=0.032), and tumor grade ( HR=1.87, 95% CI: 1.34-2.59, P=0.026) were prognostic factors for gastric cancer patients. The results of multivariate analysis showed that IBI ( HR=3.96, 95% CI: 3.51-4.48, P<0.001), T stage ( HR=2.31, 95% CI: 1.63-3.02, P=0.023), tumor grade ( HR=1.49, 95% CI: 1.12-1.83, P=0.042) were independent risk factors affecting the prognosis of gastric cancer patients. Compared with low-level IBI gastric cancer patients, the risk of death in high-level IBI patients was up to 3.96 times. Conclusion:IBI may be an important marker for diagnosis and prognosis prediction of gastric cancer, which has certain scientific and clinical value in the diagnosis and treatment of gastric cancer.
9.Research progress on the impact of lipid metabolism on endometrial receptivity and embryo implantation
Li-Na MA ; Ying QIN ; Ke-Hua WANG ; Cong-Hui PANG ; Li-Ge LU ; Wen-Xian YUAN ; Duo-Jia ZHANG ; Xiao-Ke WU
Medical Journal of Chinese People's Liberation Army 2024;49(9):1088-1093
Lipids,including fats(triglycerides)and lipoids(phospholipids and sterols),not only serve as an energy source for the body but also play a pivotal role throughout the reproductive process,particularly in the establishment and maintenance of early pregnancy.This encompasses the regulate of early embryonic development and uterine tolerance,and the facilitation of embryo implantation.Given the diversity of lipids,this review focuses on extensively studied lipid mediators such as polyunsaturated fatty acids,endocannabinoids,prostaglandins,lysophosphatidic acid,sphingolipids and steroid hormones.It systematically elaborates on the regulatory effects of fatty acid,phospholipid,and cholesterol metabolism on the formation of endometrial receptivity and embryo implantation,as well as the potential underlying mechanisms.The review aims to provide new insights and feasible intervention approaches for predicting and improving the outcomes of natural pregnancy and/or assisted reproductive technology.
10.Carrier screening for 223 monogenic diseases in Chinese population:a multi-center study in 33 104 individuals
Wei HOU ; Xiaolin FU ; Xiaoxiao XIE ; Chunyan ZHANG ; Jiaxin BIAN ; Xiao MAO ; Juan WEN ; Chunyu LUO ; Hua JIN ; Qian ZHU ; Qingwei QI ; Yeqing QIAN ; Jing YUAN ; Yanyan ZHAO ; Ailan YIN ; Shutie LI ; Yulin JIANG ; Manli ZHANG ; Rui XIAO ; Yanping LU
Journal of Southern Medical University 2024;44(6):1015-1023
Objective To investigate the epidemiological characteristics and mutation spectrum of monogenic diseases in Chinese population through a large-scale,multicenter carrier screening.Methods This study was conducted among a total of 33 104 participants(16 610 females)from 12 clinical centers across China.Carrier status for 223 genes was analyzed using high-throughput sequencing and different PCR methods.Results The overall combined carrier frequency was 55.58%for 197 autosomal genes and 1.84%for 26 X-linked genes in these participants.Among the 16 669 families,874 at-risk couples(5.24%)were identified.Specifically,584 couples(3.50%)were at risk for autosomal genes,306(1.84%)for X-linked genes,and 16 for both autosomal and X-linked genes.The most frequently detected autosomal at-risk genes included GJB2(autosomal recessive deafness type 1A,393 couples),HBA1/HBA2(α-thalassemia,36 couples),PAH(phenylketonuria,14 couples),and SMN1(spinal muscular atrophy,14 couples).The most frequently detected X-linked at-risk genes were G6PD(G6PD deficiency,236 couples),DMD(Duchenne muscular dystrophy,23 couples),and FMR1(fragile X syndrome,17 couples).After excluding GJB2 c.109G>A,the detection rate of at-risk couples was 3.91%(651/16 669),which was lowered to 1.72%(287/16 669)after further excluding G6PD.The theoretical incidence rate of severe monogenic birth defects was approximately 4.35‰(72.5/16 669).Screening for a battery of the top 22 most frequent genes in the at-risk couples could detect over 95%of at-risk couples,while screening for the top 54 genes further increased the detection rate to over 99%.Conclusion This study reveals the carrier frequencies of 223 monogenic genetic disorders in the Chinese population and provides evidence for carrier screening strategy development and panel design tailored to the Chinese population.In carrier testing,genetic counseling for specific genes or gene variants can be challenging,and the couples need to be informed of these difficulties before testing and provided with options for not screening these genes or gene variants.

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