1.Effect of The Hydrophilic Amino Acids on Self-assembly Behavior of Short Bola-like Peptides
Xin-Xin GAO ; Yu HAN ; Yi-Lin ZHOU ; Xi-Ya CHEN ; Yu-Rong ZHAO
Progress in Biochemistry and Biophysics 2025;52(5):1290-1301
ObjectiveBola-like short peptides exhibit novel self-assembly properties due to the formation of peptide dimers via hydrogen bonding interactions between their C-terminals. In this configuration, hydrophilic amino acids are distributed at both terminals, making these peptides behave similarly to Bola peptides. The electrostatic repulsive interactions arising from the hydrophilic amino acids at each terminal can be neutralized, thereby greatly promoting the lateral association of β-sheets. Consequently, assemblies with significantly larger widths are typically the dominant nanostructures for Bola-like peptides. To investigate the effect of hydrophilic amino acids on the self-assembly behavior of Bola-like peptides, the peptides Ac-RI3-CONH2 and Ac-HI3-CONH2 were designed and synthesized using the Bola-like peptide Ac-KI3-CONH2 as a template. Their self-assembly behavior was systematically examined. MethodsAtomic force microscopy (AFM) and transmission electron microscopy (TEM) were employed to characterize the morphology and size of the assemblies. The secondary structures of the assemblies were analyzed using circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy. Small-angle neutron scattering (SANS) was used to obtain detailed structural information at a short-length scale. Based on these experimental results, the effects of hydrophilic amino acids on the self-assembly behavior of Bola-like short peptides were systematically analyzed, and the underlying formation mechanism was explored. ResultsThe aggregation process primarily involved three steps. First, peptide dimers were formed through hydrogen bonding interactions between their C-terminals. Within these dimers, the hydrophilic amino acids K, R, and H were positioned at both terminals, enabling the peptides to self-assemble in a manner similar to Bola peptides. Next, β-sheets were formed via hydrogen bonding interactions along the peptide backbone. Finally, self-assemblies were generated through the lateral association of β-sheets. The results demonstrated that both Ac-KI3-CONH2 and Ac-RI3-CONH2 could self-assemble into double-layer nanotubes with diameters of approximately 200 nm. These nanotubes were formed by the edge fusion of helical ribbons, which initially emerged from twisted ribbons. Notably, the primary assemblies of these peptides exhibited opposite chirality: nanofibers formed by Ac-KI3-CONH2 displayed left-handed chirality, whereas those formed by Ac-RI3-CONH2 exhibited right-handed chirality. This reversal in torsional direction was primarily attributed to the different abilities of K and R to form hydrogen bonds with water. In contrast, Ac-HI3-CONH2 formed narrower twisted ribbons with a significantly reduced width of approximately 30 nm, which was attributed to the strong steric hindrance caused by the imidazole rings. The multilayer height of these ribbons was mainly due to the unique structure of the imidazole rings, which can function as both hydrogen bond donors and acceptors, thereby promoting aggregate growth in the vertical direction. ConclusionThe final morphology of the self-assemblies resulted from a delicate balance of various non-covalent interactions. By altering the types of hydrophilic amino acid residues in Bola-like short peptides, the relative strength of non-covalent interactions that drive assembly formation can be effectively regulated, allowing precise control over the morphology and chirality of the assemblies. This study provides a simple and effective approach for constructing diverse self-assemblies and lays a theoretical foundation for the development of functional biomaterials.
2.Tanreqing Capsules protect lung and gut of mice infected with influenza virus via "lung-gut axis".
Nai-Fan DUAN ; Yuan-Yuan YU ; Yu-Rong HE ; Feng CHEN ; Lin-Qiong ZHOU ; Ya-Lan LI ; Shi-Qi SUN ; Yan XUE ; Xing ZHANG ; Gui-Hua XU ; Yue-Juan ZHENG ; Wei ZHANG
China Journal of Chinese Materia Medica 2025;50(8):2270-2281
This study aims to explore the mechanism of lung and gut protection by Tanreqing Capsules on the mice infected with influenza virus based on "the lung-gut axis". A total of 110 C57BL/6J mice were randomized into control group, model group, oseltamivir group, and low-and high-dose Tanreqing Capsules groups. Ten mice in each group underwent body weight protection experiments, and the remaining 12 mice underwent experiments for mechanism exploration. Mice were infected with influenza virus A/Puerto Rico/08/1934(PR8) via nasal inhalation for the modeling. The lung tissue was collected on day 3 after gavage, and the lung tissue, colon tissue, and feces were collected on day 7 after gavage for subsequent testing. The results showed that Tanreqing Capsules alleviated the body weight reduction and increased the survival rate caused by PR8 infection. Compared with model group, Tanreqing Capsules can alleviate the lung injury by reducing the lung index, alleviating inflammation and edema in the lung tissue, down-regulating viral gene expression at the late stage of infection, reducing the percentage of neutrophils, and increasing the percentage of T cells. Tanreqing Capsules relieved the gut injury by restoring the colon length, increasing intestinal lumen mucin secretion, alleviating intestinal inflammation, and reducing goblet cell destruction. The gut microbiota analysis showed that Tanreqing Capsules increased species diversity compared with model group. At the phylum level, Tanreqing Capsules significantly increased the abundance of Firmicutes and Actinobacteria, while reducing the abundance of Bacteroidota and Proteobacteria to maintain gut microbiota balance. At the genus level, Tanreqing Capsules significantly increased the abundance of unclassified_f_Lachnospiraceae while reducing the abundance of Bacteroides, Eubacterium, and Phocaeicola to maintain gut microbiota balance. In conclusion, Tanreqing Capsules can alleviate mouse lung and gut injury caused by influenza virus infection and restore the balance of gut microbiota. Treating influenza from the lung and gut can provide new ideas for clinical practice.
Animals
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Drugs, Chinese Herbal/administration & dosage*
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Mice
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Lung/metabolism*
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Mice, Inbred C57BL
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Capsules
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Orthomyxoviridae Infections/virology*
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Gastrointestinal Microbiome/drug effects*
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Male
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Humans
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Female
;
Influenza A virus/physiology*
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Influenza, Human/virology*
3.Clinical efficacy of multi-technique combination in the treatment of ischemic diabetic foot.
Hui-Yan LIU ; Yi YOU ; Wen-Gao WU ; Sheng RONG ; Qing-Hua ZHOU ; Na-Xin ZENG
China Journal of Orthopaedics and Traumatology 2025;38(9):917-923
OBJECTIVE:
To explore clinical efficacy of different technical combinations in treating ischemic diabetic foot (DF).
METHODS:
A retrospective analysis was conducted on 35 patients with DF who were treated with vascular interventional opening technique, periosteal distraction technique and bone cement coverage technique from January 2024 to November 2024. They were divided into comprehensive group and periosteal distraction group according to whether the vascular interventional opening technique was used in combination or not. There were 5 patients in comprehensive group, including 4 males and 1 female, aged from 59 to 73 years old with an average of (64.40±5.46) years old;the duration of diabetes ranged from 0.17 to 30.00 years with an average of (14.63±12.02) years;the courses of DF ranged from 30 to 150 days with an average of (84.00±61.48) days;2 patients were grade 2, 2 patients were grade 3, and 1 patient was grade 4 according to Wagner classification;combined vascular interventional opening, periosteal distraction and bone cement coverage surgery for treatment. There were 30 patients in periosteal stretch group, including 22 males and 8 females, aged from 58 to 86 years old with an average of (72.63±7.84) years old;the duration of diabetes was 10.00 (6.75, 16.75) years;the courses of DF was 30.00 (15.00, 37.50) days;14 patients were grade 2, 11 patients were grade 3, and 5 patients were grade 4 according to Wagner classification; combined periosteal distraction and bone cement coverage surgery for treatment. Changes of C-reactive protein (CRP), interleukin-6 (IL-6), and procalcitonin (PCT), toe skin temperature, peripheral capillary oxygen saturation (SpO2), and visual analogue scale (VAS) for pain were compared between two groups before operation and 1 week after operation. The number of operations, healing period, healing number, toe amputation number, preoperative fever situation and the number of complications were compared between two groups.
RESULTS:
Both groups were followed up for at least 6 months. There were no statistically significant differences in the number of operations, healing period, toe amputation rate, wound healing rate and complications between two groups (P>0.05). Before operation, the toe skin temperature of comprehensive group (26.98±0.88) ℃ was lower than that of periosteal distraction group (28.17±1.45) ℃, and the difference was statistically significant (P<0.05);while there were no statistically significant difference in CRP, IL-6, PCT, toe SpO2 and VAS between two groups (P>0.05). At 1 week after operation, IL-6, toe skin temperature, toe SpO2 and VAS in comprehensive group were 12.29(7.92, 22.15) pg·ml-1, (36.02±0.23) ℃, (95.80±0.84) % and(1.40±0.55) respectively, while those in periosteal distraction group were 5.49(4.36, 7.45) pg·ml-1, (31.36±1.57) ℃, (84.53±6.38) %, (2.20±0.81);and there were statistically significant differences between two groups(P<0.05). CRP, IL-6 and VAS at 1 week after operation in both groups were decreased compared with those before operation, and the differences were statistically significant(P<0.05). The toe skin temperature and SpO2 were increased compared with those before operation, and the differences were statistically significant(P<0.001).
CONCLUSION
The multi-technique combination therapy, including vascular interventional opening technique, periostealdistraction technique and bone cement covering technique, could protect each other, enhance efficacy, effectively promote the wound healing of ischemic diabetic foot ulcer, and reduce the toe amputation rate. For moderate to severe ischemic DF, the combined use of periosteal distraction and bone cement coverage techniques has a satisfactory effect. For extremely severe ischemic DF with inflow tract lesions, vascular interventional opening techniques need to be added.
Humans
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Male
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Female
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Middle Aged
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Aged
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Diabetic Foot/surgery*
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Retrospective Studies
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Aged, 80 and over
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Ischemia/surgery*
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Interleukin-6/metabolism*
4.Body fat distribution and semen quality in 4304 Chinese sperm donors.
Si-Han LIANG ; Qi-Ling WANG ; Dan LI ; Gui-Fang YE ; Ying-Xin LI ; Wei ZHOU ; Rui-Jun XU ; Xin-Yi DENG ; Lu LUO ; Si-Rong WANG ; Xin-Zong ZHANG ; Yue-Wei LIU
Asian Journal of Andrology 2025;27(4):524-530
Extensive studies have identified potential adverse effects on semen quality of obesity, based on body mass index, but the association between body fat distribution, a more relevant indicator for obesity, and semen quality remains less clear. We conducted a longitudinal study of 4304 sperm donors from the Guangdong Provincial Human Sperm Bank (Guangzhou, China) during 2017-2021. A body composition analyzer was used to measure total and local body fat percentage for each participant. Generalized estimating equations were employed to assess the association between body fat percentage and sperm count, motility, and morphology. We estimated that each 10% increase in total body fat percentage (estimated change [95% confidence interval, 95% CI]) was significantly associated with a 0.18 × 10 6 (0.09 × 10 6 -0.27 × 10 6 ) ml and 12.21 × 10 6 (4.52 × 10 6 -19.91 × 10 6 ) reduction in semen volume and total sperm count, respectively. Categorical analyses and exposure-response curves showed that the association of body fat distribution with semen volume and total sperm count was stronger at higher body fat percentages. In addition, the association still held among normal weight and overweight participants. We observed similar associations for upper limb, trunk, and lower limb body fact distributions. In conclusion, we found that a higher body fat distribution was significantly associated with lower semen quality (especially semen volume) even in men with a normal weight. These findings provide useful clues in exploring body fat as a risk factor for semen quality decline and add to evidence for improving semen quality for those who are expected to conceive.
Humans
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Male
;
Adult
;
Semen Analysis
;
China
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Body Fat Distribution
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Longitudinal Studies
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Sperm Count
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Sperm Motility
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Body Mass Index
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Tissue Donors
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Obesity/complications*
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Spermatozoa
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Young Adult
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Middle Aged
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East Asian People
5.Two new isocoumarins from cave-derived Metarhizium anisopliae NHC-M3-2
Rong-xiang WU ; Jie-lan GUO ; Hua HUANG ; Jing-jing LIAO ; Yi HAO ; Fan-dong KONG ; Li-man ZHOU ; Chao-jun ZHANG
Acta Pharmaceutica Sinica 2024;59(9):2588-2593
Extracting extracts of secondary metabolites from the karst cave fungus
6.The value of ultrasonography combined with serum thyroglobulin levels in preoperative N staging and therapeutic effect evaluation in patients with thyroid cancer
Rong ZHOU ; Yi LIU ; Xiuhua YU
Chinese Journal of Primary Medicine and Pharmacy 2024;31(3):366-370
Objective:To investigate the clinical value of ultrasonography combined with serum thyroglobulin (Tg) levels in preoperative N staging and therapeutic effect evaluation in patients with thyroid cancer.Methods:A retrospective analysis was conducted on the clinical data of 125 patients with thyroid cancer who underwent total thyroidectomy at The 903 Hospital of PLA Joint Logistics Support Force between January 2016 and December 2021. Based on the pathological results, the patients were divided into stages N0, N1a, and N1b. Consistency testing was performed to assess the consistency between ultrasonographic and pathological findings. The serological markers were compared between stage N0 and stage N1. A multivariate logistic regression analysis was conducted to investigate the correlation between antithyroid peroxidase antibodies (TPOAb) and Tg and lymph node metastasis. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of serum Tg, ultrasonography, and the combination of serum Tg with ultrasonography for lymph node metastasis.Results:Among the 125 patients, 51 cases were diagnosed with stage N1, including 36 cases of stage N1a and 15 cases of stage N1b. The pathological examination confirmed stage N0 in 74 cases. Ultrasonography results showed stage N0 in 96 cases, stage N1a in 7 cases, and stage N1b in 22 cases. The preoperative N staging of thyroid cancer patients by ultrasonography was moderately consistent with the pathological staging ( Kappa = 0.44, P < 0.001). Serum free triiodothyronine and Tg levels were higher in stage N1 compared with stage N0 ( P < 0.001), while serum TPOAb levels were lower in stage N1 than in stage N0 ( P = 0.017). Multivariate logistic regression analysis was performed with lymph node metastasis as the dependent variable and the factors FT3, TPOAb, and Tg levels as independent variables. The analysis revealed that Tg was associated with lymph node metastasis ( OR = 1.02, P < 0.001). The ROC curve analysis showed that the area under the ROC curve for serum Tg level and ultrasonography for the diagnosis of lymph node metastasis were 0.67 and 0.65, respectively, which were significantly lower than the area under the curve (0.76) for the combined detection ( P < 0.001). Conclusion:The combined utilization of ultrasonography and serum Tg levels in the preoperative staging of thyroid cancer patients holds distinct clinical relevance. Notably, this combined approach offers a more precise diagnosis of lymph node metastasis compared with ultrasonography alone. Consequently, when making clinical judgments regarding preoperative lymph node metastasis, it is advisable to consider the results of both ultrasonography and serum Tg testing to facilitate surgical planning and evaluate surgical outcomes.
7.Efficacy of Fuzheng Hejie Prescription in the Treatment of Respiratory Viral Infection in Children and Its Effect on Immune Function
Xin-Yi LI ; Zong-Kan HU ; Yu XIE ; Wen-Ting MA ; Rong-Fang ZHOU ; Qi LYU ; Jie-Yu ZAN ; Ling-Fang ZHOU ; Ze-Ting YUAN
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(3):631-637
Objective To investigate the efficacy of Fuzheng Hejie Prescription(composed of Scutellariae Radix,Lonicerae Japonicae Flos,Agastachis Herba,Bupleuri Radix,Atractylodis Rhizoma,Glycyrrhizae Radix et Rhizoma,etc.)in the treatment of respiratory viral infections in children and to observe its effect on inflammatory factors and immune function.Methods A total of 203 children with respiratory viral infection of H1N1 virus were randomly divided into 101 cases in the observation group and 102 cases in the control group.Both groups were given the routine treatment for subsiding fever,maintaining water-electrolyte balance,and ensuring enough sleep.And additionally,the control group was given Ribavirin Granules and Ibuprofen Granules,and the observation group was given Fuzheng Hejie Prescription based on the treatment for the control group.The course of treatment covered 7 days.The changes of traditional Chinese medicine(TCM)syndrome scores and the levels of immunological indicators and inflammatory factors in the two groups were observed before and after the treatment.Moreover,the clinical efficacy,symptom resolution time and the incidence of adverse reactions were compared between the two groups of children.Results(1)In the course of the trial,one case fell off in the observation group and 2 cases fell off in the control group,and eventually 100 children in each group were included in the trial.(2)After 7 days of treatment,the total effective rate of the observation group was 93.00%(93/100),and that of the control group was 88.00%(88/100),and the intergroup comparison showed that the therapeutic effect of the observation group was superior to that of the control group,but the difference was not statistically significant(χ2= 1.454,P = 0.228).(3)After treatment,the scores of primary symptoms and secondary symptoms as well as the total TCM syndrome scores in the two groups were decreased compared with those before treatment(P<0.05),and the decrease in the observation group was significantly superior to that in the control group(P<0.01).(4)After treatment,the time for the resolution of clinical symptoms such as fever,cough,expectoration and sore throat in the observation group was significantly shorter than that in the control group(P<0.01).(5)After treatment,the levels of immunological indicators of T lymphocyte subset CD3+ and CD4+ in the two groups were increased compared with those before treatment(P<0.05),and the levels of CD8+ and B cells were decreased compared with those before treatment(P<0.05).The intergroup comparison showed that the increase in the levels of CD3+ and CD4+ as well as the decrease in the levels of CD8+ and B cells of the observation group was significantly superior to that of the control group(P<0.01).(6)After treatment,the levels of inflammatory factors of serum amyloid A(SAA),C-reactive protein(CRP),serum tumor necrosis factor alpha(TNF-α),soluble interleukin 2 receptor(SIL-2R),and interleukin 6(IL-6)in the two groups were significantly decreased compared with those before treatment(P<0.05),and the levels of interleukin 2(IL-2)and interferon γ(IFN-γ)ls were all significantly increased compared with those before treatment(P<0.05).The intergroup comparison showed that the decrease of serum SAA,CRP,TNF-α,SIL-2R,and IL-6 levels and the increase of serum IL-2 and IFN-γ levels in the observation group were significantly superior to those in the control group(P<0.01).(7)The incidence of adverse reactions in the observation group was 2.00%(2/100),which was significantly lower than that of 8.00%(8/100)in the control group,but the difference was not statistically significant(χ2 = 3.789,P = 0.052).Conclusion Fuzheng Hejie Prescription exerts certain effect in treating children with respiratory viral infection of H1N1 virus,which can effectively decrease children's TCM syndrome scores,regulate the inflammatory response,improve the immune function,accelerate the relief of clinical symptoms and shorten the course of the disease.
8.Clinical Observation on the Therapeutic Effect of Lingnan Traditional Vesiculating Moxibustion No.4 Recipe in the Treatment of Mild Depressive Disorder
Yi-Rong ZHOU ; Hui FANG ; Wan-Xin ZHANG ; Xiang JI ; Wen-Bin FU
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(4):959-964
Objective To observe the clinical efficacy of Lingnan Traditional Vesiculating Moxibustion No.4 Recipe(mainly composed of Brassicae Junceae Semen,Euodiae Fructus,and Curcumae Radix)in the treatment of mild depressive disorder(DD),and to provide a novel approach to the treatment of mild DD population.Methods Sixty-one patients with mild DD were randomly divided into 31 cases in the trial group and 30 cases in the control group.The trial group was given medicinal vesiculation treatment with Lingnan Traditional Vesiculating Moxibustion No.4 Recipe,and the control group was given medicinal vesiculation treatment with the placebo of Lingnan Traditional Vesiculating Moxibustion No.4 Recipe.The treatment was performed twice a week and with an interval of 3-4 days between the treatment,and the course of treatment covered 6 weeks.The changes of Hamilton Depression Scale-17(HAMD-17)scores and Patient Health Questionnaire-9(PHQ-9)scores in the two groups were observed before and after the treatment.After treatment,the clinical efficacy and safety of the patients in the two groups were evaluated.Results(1)After 6 weeks of treatment,the total efficacy rate of the trial group was 77.42%(24/31),and that of the control group was 26.67%(8/30).The intergroup comparison(tested by rank sum test)showed that the efficacy of the trial group was significantly superior to that of the control group,and the difference was statistically significant(P<0.01).(2)After treatment,the HAMD-17 scores and PHQ-9 scores of patients in the two groups were lower than those before treatment(P<0.01),and the decrease of HAMD-17 and PHQ-9 scores in the trial group was significantly superior to that of the control group,the difference being statistically significant(P<0.01).(3)During the trial,there were 5 cases of adverse events related to the vesiculating moxibustion treatment,and all 5 cases of adverse events occurred in the trial group,manifested as minor blisters at the acupoint application region.The 5 cases kept on participating in the trial after relevant treatment.Conclusion Lingnan Traditional Vesiculating Moxibustion No.4 Recipe can effectively relieve the clinical symptoms of patients with mild DD,and has high safety.
9.Clinical observation of splenectomy with distal pancreatectomy during cytoreductive surgery in epithelial ovarian cancer
Yi-Xuan LIU ; Qian-Qian YAN ; Yu-Lian CHEN ; Ying ZHOU ; Rong JIANG
Fudan University Journal of Medical Sciences 2024;51(1):50-55
Objective To evaluate the safety and efficacy of splenectomy with distal pancreatectomy during cytoreductive surgery in epithelial ovarian cancer(EOC).Methods A total of 17 patients from Zhongshan Hospital,Fudan University and the First Affiliated Hospital of University of Science and Technology of China(Anhui Provincial Hospital)received splenectomy with distal pancreatectomy during cytoreductive surgery in EOC were recruited.Their clinicopathological characteristics,postoperative complications and survival situation were retrospective analyzed.Results Of the 17 patients,there were 13 primary cases and 4 recurrent cases.Eleven cases(64.7%)had preoperative imaging finding with metastatic lesions in the splenic hilum,among whom 6 cases had distal pancreas metastasis during the operation.The drainage was placed in the splenic fossa for the measurement of amylase levels in drain fluid and was removed after 8(3-12)days.There were 4 patients had postoperative pancreatic fistula(POPF)of grade A,3 patients had POPF of grade B and no POPF of grade C occurred.The 2 patients with POPF of grade B improved after percutaneous drainage,and the rest recovered with somatostatin,antibiotic drugs and medicines without perioperative mortality.The interval between surgery to chemotherapy was 17.5(13-37)days.The median follow-up time was 14(4-64)months and the median progression-free survival was 10(5-32)months.Conclusion Splenectomy with distal pancreatectomy as part of cytoreduction surgery in EOC is needed for optimal resection,and the complication of pancreatic fistula could be managed conservatively.
10.The relationship between activities of daily living and mental health in community elderly people and the mediating role of sleep quality
Heng-Yi ZHOU ; Jing LI ; Dan-Hua DAI ; Yang LI ; Bin ZHANG ; Rong DU ; Rui-Long WU ; Jia-Yan JIANG ; Yuan-Man WEI ; Jing-Rong GAO ; Qi ZHAO
Fudan University Journal of Medical Sciences 2024;51(2):143-150
Objective To explore the relationship and internal path between activities of daily living(ADL),sleep quality and mental health of community elderly people in Shanghai.Methods A questionnaire survey was conducted among community residents aged 60 years and older seeing doctors in community health care center of five streets in Shanghai during Sept to Dec,2021 using convenience sampling.Activities of Daily Living(ADL),Pittsburgh Sleep Quality Index(PSQI)and 10-item Kessler Psychological Distress Scale(K10)were adopted in the survey.Single factor analysis,correlation analysis and multiple linear regression were used to analyze the data.The effect relationship between the variables was tested using Bootstrap's mediated effects test.Results A total of 1 864 participants were included in the study.The average score was 15.53±4.47 for ADL,5.60±3.71 for PSQI and 15.50±6.28 for K10.The rate of ADL impairment,poor sleep quality,poor and very poor mental health of the elderly were 23.6%,27.3%,11.9%and 4.9%,respectively.ADL and sleep quality were all positively correlated with mental health(r=0.321,P<0.001;r=0.466,P<0.001);ADL was positively correlated with sleep quality(r=0.294,P<0.001).Multiple linear results of factors influencing mental health showed that ADL(β= 0.457,95%CI:0.341-0.573),sleep quality(β =0.667,95%CI:0.598-0.737)and mental health were positively correlated(P<0.001).Sleep quality partially mediated the relationship between ADL and mental health(95%CI:0.078-0.124)with an effect size of 33.0%.Conclusion Sleep quality is a mediator between ADL and mental health among community elderly people.Improving ADL and sleep quality may improve mental health in the population.

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