OBJECTIVE: To explore the role of the natural compound hesperidin in glycolysis, the key ratelimiting enzyme, in colorectal cancer (CRC) cell lines. METHODS: In vitro, HCT116 and SW620 were treated with different doses of hesperidin (0-500 µmol/L), cell counting kit-8 and colone formation assays were utilized to detected inhibition effect of hesperidin on CRC cell lines. Transwell and wound healing assays were performed to detect the ability of hesperidin (0, 25, 50 and 75 µmol/L) to migrate CRC cells. To confirm the apoptotic-inducing effect of hesperidin, apoptosis and cycle assays were employed. Western blot, glucose uptake, and lactate production determination measurements were applied to determine inhibitory effects of hesperidin (0, 25 and 50 µmol/L) on glycolysis. In vivo, according to the random number table method, nude mice with successful tumor loading were randomly divided into vehicle, low-dose hesperidin (20 mg/kg) and high-dose hesperidin (60 mg/kg) groups, with 6 mice in each group. The body weights and tumor volumes of mice were recorded during 4-week treatment. The expression of key glycolysis rate-limiting enzymes was determined using Western blot, and glucose uptake and lactate production were assessed. Finally, protein interactions were probed with DirectDIA Quantitative Proteomics, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. RESULTS: Hesperidin could inhibit CRC cell line growth (P<0.05 or P<0.01). Moreover, hesperidin presented an inhibitory effect on the migrating abilities of CRC cells. Hesperidin also promoted apoptosis and cell cycle alterations (P<0.05). The immunoblotting results manifested that hesperidin decreased the levels of hexokinase 2, glucose transporter protein 1 (GLUT1), GLUT3, L-lactate dehydrogenase A, 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 2 (PFKFB2), PFKFB3, and pyruvate kinase isozymes M2 (P<0.01). It remarkably suppressed tumor xenograft growth in nude mice. GO and KEGG analyses showed that hesperidin treatment altered metabolic function. CONCLUSION Hesperidin inhibits glycolysis and is a potential therapeutic choice for CRC treatment.
Hesperidin/therapeutic use* ; Colorectal Neoplasms/metabolism* ; Glycolysis/drug effects* ; Animals ; Humans ; Apoptosis/drug effects* ; Mice, Nude ; Cell Movement/drug effects* ; Cell Line, Tumor ; Cell Proliferation/drug effects* ; Glucose/metabolism* ; Cell Cycle/drug effects* ; Mice, Inbred BALB C ; Mice ; HCT116 Cells ; Lactic Acid

Objective To explore the clinical significance of serum cytokine expression in the hepatitis B surface antigen(HBsAg)sero-clearance in patients with severe hepatitis B.Methods A nested case-control trial was conducted on 14 inpatients with severe hepatitis B admitted in our hospital from 2006 to 2020.Of them,7 patients(aged 36.57±3.15 years)achieved HBsAg sero-clearance within 1 year after the onset of hepatitis B flares(with abrupt rise of ALT level to＞5 times the upper limit of normal during HBV infection)and were assigned into HBsAg clearance group,while the other 7 patients(aged 34.14±2.97 years)only obtained HBsAg decreased less than 1 g within 1 year after the onset(HBsAg non-clearance group).Then,multiplex liquid-chip assay based on Luminex xMAP was used to detect the expression levels of 48 cytokines such as IFN-γ and IL-2 in serum samples of these 14 patients.Results The serum levels IFN-γ,IL-2Ra and SDF-1α were significantly lower in the HBsAg clearance group than the HBsAg non-clearance group(P＜0.05),but no statistical differences were observed in other 39 cytokines between the 2 groups.And there were 5 cytokines having no mutual expression in both groups.The copy number of HBV DNA was positively correlated with serum HGF(r=0.675,P=0.008)and SDF-1α levels(r=0.587,P=0.027),while negatively with IP-10(r=-0.600,P=0.023)and MIG level(r=-0.640,P=0.014).Meanwhile,a positive correlation was found between HBsAg titer and IL12-p70 level(r=0.593,P=0.025),and a negative correlation between HBsAg titer and TNF-α level(r=-0.609,P=0.021).In addition,the serum total bilirubin level was positively correlated with the expression of SCGF-β(r=0.543,P=0.045).Conclusion Three differentially expressed cytokines and some cytokines related to HBV DNA level and HBsAg titer are found,which may provide new insights into the underlying immunological mechanism of HBV virus clearance caused by hepatitis flares.Meanwhile,it also provides potential biomarkers for HBsAg serological clearance in patients with severe hepatitis B.

Objective: To investigate surgical strategies and the corresponding benefits for patients with perihilar cholangiocarcinoma(pCCA). Methods: A total of 81 patients with pCCA who underwent radical excision in the Department of Biliary and Pancreatic Surgery of Sun Yat-Sen Memorial Hospital between January 2014 and December 2021 were retrospectively collected.The cohort consisted of 50 male and 31 female patients,with an age of (62.5±11.5)years(range:26 to 83 years).Seventy-five cases were diagnosed with jaundice,60 of whom received preoperative biliary drainage,while 20 patients received portal vein embolization.Their serum bilirubin level within one week before the operation(M(IQR)) was 44.3 (41.9) μmol/L(range:8.0 to 344.2 μmol/L).Preoperative imaging examinations were performed to evaluate the Bismuth-Corlette type of pCCA,showing 3,6,21,27,and 24 cases of Bismuth-Corlette type Ⅰ,Ⅱ,Ⅲa,Ⅲb,and Ⅳ,respectively.The primary outcome was overall survival (OS),and the secondary outcomes were relapse-free survival (RFS),90-day postoperative morbidity and 90-day postoperative mortality.OS and RFS were estimated using the Kaplan-Meier method and compared by the Log-rank test.Significant prognostic factors were determined using univariate and multivariable Cox proportional hazard regression analyses. Results: In the cohort of 81 pCCA patients,67 cases(82.7%) underwent major hepatectomy while 3 cases received major hepatectomy combined with pancreaticoduodenectomy.Thirty-four patients underwent hepatectomy combined with vascular resection and reconstruction(18 cases of portal vein resection and reconstruction alone;9 cases of hepatic artery resection and reconstruction alone;7 cases of combination of portal vein and hepatic artery resection and reconstruction).Margin negative(R0 excision) were achieved in 53.1%(43/81) of these patients.The operation duration was (627±136)minutes(range:565 to 940 minutes),and the intraoperative blood loss was 400(455)ml(range:200 to 2 800 ml).The 90-day postoperative mortality was 3.7%(3/81).Grade 3-4 postoperative morbidity was 23.4% (19/81) according to the Clavien-Dindo classification of surgical complications.Up to the last follow-up at September 2022,the follow-up time was 34.0(24.2)months (range:0.4 to 103.6 months).Three patients who died within 90 days after surgery were excluded from the survival analysis.The median OS was 36.10 months (95%CI:18.23 to 42.97 months) and the 1-,3-and 5-year OS rates were 85.3%,46.8% and 27.3%,respectively.The median OS of 41 patients with negative margins was 47.83 months(95%CI:36.90 to 58.80 months) and that of 37 patients with positive margins was 20.47 months(95%CI:10.52 to 30.58 months).The median RFS of 70 patients with R0 and R1 resection was 24.50 months(95%CI:12.15 to 31.85 months)and the 1-,3-and 5-year RFS rates were 65.2%,45.7% and 29.9%,respectively.The median RFS of 41 patients with R0 resection was 38.57 months(95%CI:21.50 to 55.63 months) and that of 29 patients with R1 resection was 10.83 months(95%CI:2.82 to 19.86 months). Conclusions: The primary therapy for pCCA is radical surgical resection.A precise preoperative evaluation and sufficient preparation can reduce postoperative morbidity.Surgical treatment can achieve a better survival outcome by increasing the radical resection rate.

There has been increasing interest in the psycho-socio-relational and sexual disorders of infertility, as the risk of psychological burden among infertile men with sexual dysfunctions is significant. The purpose of this study was to develop and to validate a predictive model to estimate individual psychological burden among infertile men with sexual dysfunction and study the association between them. Comprehensive data were collected for infertile men (n = 480) who sought treatment for infertility in a reproductive medicine center between June 2012 and December 2013. Using independent predictors of psychological burden from the least absolute shrinkage and selection operator, univariable and multivariable analyses were developed into two models. Predictive accuracy was compared between the models. We explored the association between sexual dysfunction and psychological burden. A total of 480 patients were analyzed using 10-fold cross-validation. Independent predictors of psychological burden were incorporated into a model to measure anxiety (corrected-area under curve (AUC): 77.3%) and a model to measure depression (corrected-AUC: 70.2%). Anxiety and depression were both associated with erectile dysfunction (P < 0.05), with anxiety demonstrating the strongest association. Only anxiety was associated with premature ejaculation (P < 0.05). Premature ejaculation was not found to be associated with depression (P > 0.05). Predictive models for psychological burden among infertile men with sexual dysfunction are presented, and we found that there is an association between psychological burden and sexual dysfunction. According to the models, proper counseling and treatment of sexual dysfunction in infertile men may reduce the psychological burden, help attain natural pregnancy, and improve the quality of life.
Adolescent ; Adult ; Anxiety/psychology* ; Cross-Sectional Studies ; Depression/psychology* ; Erectile Dysfunction/psychology* ; Humans ; Infertility, Male/psychology* ; Male ; Quality of Life/psychology* ; Surveys and Questionnaires ; Young Adult

ObjectiveTo investigate the protective effect of human urine-derived stem cells (USCs) on erectile function and cavernous structure in rats with cavernous nerve injury (CNI).

METHODSSixty adult male SD rats with normal sexual function were randomly divided into four groups of equal number: sham operation, bilateral CNI (BCNI) model control, phosphate buffered saline (PBS), and USC. The BCNI model was established in the latter three groups of rats by clamping the bilateral cavernous nerves. After modeling, the rats in the PBS and USC groups were treated by intracavernous injection of PBS at 200 μl and USCs at 1×106/200 μl PBS respectively for 28 days. Then, the maximum intracavernous pressure (mICP) and the ratio of mICP to mean arterial pressure (mICP/MAP) of the rats were calculated by electrical stimulation of the major pelvic ganglions, the proportion of nNOS- or NF200-positive nerve fibers in the total area of penile dorsal nerves determined by immunohistochemical staining, the levels of endothelial cell marker eNOS, smooth muscle marker α-SMA and collagen I detected by Western blot, and the smooth muscle to collagen ratio and the cell apoptosis rate in the corpus cavernosum measured by Masson staining and TUNEL, respectively.

RESULTSAfter 28 days of treatment, the rats in the USC group, as compared with those in the PBS and BCNI model control groups, showed significant increases in the mICP (［81 ± 9.9］ vs ［31 ± 8.3］ and ［33 ± 4.2］ mmHg, P <0.05), mICP/MAP ratio (0.72 ± 0.05 vs 0.36 ± 0.03 and 0.35 ± 0.04, P <0.05), the proportions of nNOS-positive nerve fibers (［11.31 ± 4.22］% vs ［6.86 ± 3.08］% and ［7.29 ± 4.84］% , P <0.05) and NF200-positive nerve fibers in the total area of penile dorsal nerves (［27.31 ± 3.12］% vs ［17.38 ± 2.87］% and ［19.49 ± 4.92］%, P <0.05), the eNOS/GAPDH ratio (0.52 ± 0.08 vs 0.31 ± 0.06 and 0.33 ± 0.07, P <0.05), and the α-SMA/GAPDH ratio (1.01 ± 0.09 vs 0.36 ± 0.05 and 0.38 ± 0.04, P <0.05), but a remarkable decrease in the collagen I/GAPDH ratio (0.28 ± 0.06 vs 0.68 ± 0.04 and 0.70 ± 0.10, P <0.05). The ratio of smooth muscle to collagen in the corpus cavernosum was significantly higher in the USC than in the PBS and BCNI model control groups (17.91 ± 2.86 vs 7.70 ± 3.12 and 8.21 ± 3.83, P <0.05) while the rate of cell apoptosis markedly lower in the former than in the latter two (3.31 ± 0.83 vs 9.82 ± 0.76, P <0.01; 3.31 ± 0.83 vs 9.75 ± 0.91, P <0.05).

CONCLUSIONSIntracavernous injection of USCs can protect the erectile function of the rat with cavernous nerve injury by protecting the nerves, improving the endothelial function, alleviating fibrosis and inhibiting cell apoptosis in the cavernous tissue.

Actins ; analysis ; Animals ; Arterial Pressure ; Collagen ; analysis ; Disease Models, Animal ; Erectile Dysfunction ; prevention & control ; Male ; Nitric Oxide Synthase Type I ; analysis ; Nitric Oxide Synthase Type III ; analysis ; Penile Erection ; physiology ; Penis ; innervation ; Pudendal Nerve ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Saline Solution ; administration & dosage ; Stem Cell Transplantation ; methods ; Stem Cells ; Urine ; cytology

Objective: To explore the clinical efficacy and safety of unrelated umbilical cord blood transplantation (UCBT) in the treatment of refractory and relapsed acute leukemia (AL) patients. Methods: The clinical data of 22 refractory and relapsed AL patients who were treated with UCBT as salvage therapy from November 2009 to May 2017 were retrospectively analyzed. All patients received a myeloablative conditioning regimen for prevention of graft-versus-host disease (GVHD) with cyclosporine A (CSA)/short course of mycophenolate mofetil (MMF). Results: ①Of 22 patients, 9 cases were male and 13 female. The median age was 23 (15-44) years and median weight of 52.5 (43-82) kg. All patients were transplanted with a median umbilical cord blood nucleated cells of 3.07 (1.71-5.30)×10⁷/kg (by weight), the median CD34⁺ cells was 1.60 (0.63-3.04)×10⁵/kg (by weight). ②The myeloid cumulative implantation rate was 95.5% (95%CI 45.2-99.7%) after transplantation of 42 d, with the median implantation time of 19 (13-27) d. The platelet cumulative implantation rate after transplantation of 120 d was 81.8% (95%CI 54.2-93.6%), the median implantation time of 42 (20-164) d. ③The incidence of Ⅱ-Ⅳ, Ⅲ-Ⅳ aGVHD and the 2 year cumulative incidence of cGVHD were 36.4%, 13.6% and 40.3% respectively. ④ The transplant related mortality (TRM) after transplantation of 180d was 22.7%, 2 year cumulative rate of relapse was 18.7% (95%CI 3.6-42.5%), 2 year disease-free survival rate (DFS) and overall survival rate (OS) were 53.7% and 58.1%, respectively. Conclusion: The preliminary results show that the use of UCBT is safe and effective for refractory and relapsed AL patients who fail to respond to conventional chemotherapy.
Acute Disease ; Adolescent ; Adult ; Cord Blood Stem Cell Transplantation ; Female ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia/therapy* ; Male ; Peripheral Blood Stem Cell Transplantation ; Retrospective Studies ; Transplantation Conditioning ; Young Adult

Objective To assess the peripheral arterial structural and elastic remodeling in patients with lower extremity atherosclerotic disease (LEAD).Methods Thirty-one patients with LEAD and 34 age-,sex-matched healthy subjects (control group) were enrolled in this study.The intima-media thickness (IMT),diameter (D) and parameters of arterial stiffness (β,pulse wave velocity (PWVβ)) were measured by acquiring the longitudinal view of left and right common femoral artery (LCFA,RCFA) and common carotid artery (LCCA,RCCA) by quality intima-media thickness (QIMT) and quality arterial stiffness (QAS) techniques.The factors correlated with arterial stiffness in LEAD patients were analyzed.Results ① The systolic blood pressure(SBP),pulse pressure(PP),smoking pack-year and smoking extent (nonsmoker,smoker with ＜40 pack-year,or smoker with ≥40 pack-year) were significantly higher in LEAD group than those in the control group (P ＜0.05).②IMT of the LCCA and the diameters (D) of LCCA and RCCA were significantly increased in the LEAD group (P ＜0.05-0.01).As to the both sides CFA,IMT and IMT/D values were significantly increased (P ＜ 0.01).The mean IMT (mIMT) of both sides of CCA and CFA were increased (P ＜0.05).The values of β,PWVβ of LCCA and PWVβ of RCCA were significantly increased,and β and PWVβ values of both sides of CFA were significantly increased in the LEAD group (P ＜ 0.05).There was a positive correlation between the stiffness indexes of the carotid artery and that of the femoral artery (P ＜0.01).③The femoral mβ was correlated with femoral mIMT,SBP,PP,smoking amount and smoking extent,and the femoral mPWVβ was also correlated with mIMT,age,SBP,PP,smoking pack-year and smoking extent (P ＜0.05-0.01).In multivariable stepwise regression analysis,mIMT and PP were factors independently correlated with femoral mβ.And SBP,smoking extent,and mIMT were independently correlated with femoral mPWVβ.Conclusions Peripheral arteries in patients with LEAD experience vascular remodeling,as well as increased carotid and femoral stiffness.Femoral stiffness is correlated with IMT,arterial blood pressure and smoking extent.

OBJECTIVETo observe the efficacy of the combination of Qilin Pills and levofloxacin in the treatment of asthenospermia accompanied with accessory sex gland infection.

METHODSWe randomly assigned 80 asthenospermia patients with accessory sex gland infection to groups 1 and 2 of equal number, the former treated with Qilin Pills + levofloxacin, and the latter with levofloxacin only. Qilin Pills were administered at the dose of 6 g tid for 30 days, and levofloxacin at the dose of 0.5 g qd for 20 days. We obtained semen parameters, including the percentage of progressively motile sperm and peroxidase-positive white blood cell (WBC) count, before and after medication, and compared the clinical effects between the two groups.

RESULTSAll the patients accomplished the clinical trial. The therapeutic effectiveness rates in improving progressive sperm motility were 60% in group 1 and 17.5% in group 2, with statistically significant differences between the two groups (P < 0.05). As for the peroxidase-positive WBC count in semen, the effectiveness rates were 87.5% and 82.5%, respectively, with no significant differences between the two groups (P > 0.05).

CONCLUSIONFor the treatment of asthenospermia accompanied with accessory sex gland infection, Qilin Pills combined with levofloxacin is evidently better than levofloxacin alone in improving sperm motility, and it has no obvious adverse effects.

Adult ; Asthenozoospermia ; complications ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Infertility, Male ; drug therapy ; Levofloxacin ; therapeutic use ; Male ; Phytotherapy ; Reproductive Tract Infections ; drug therapy ; Treatment Outcome ; Young Adult

Objective To evaluate the correlation between left ventricular function and arterial stiffness of left femoral artery in patients with lower extremity atherosclerotic disease (LEAD).Methods Thirty-three patients with LEAD and 37 healthy subjects (control group) were enrolled in this study.The intima-media thickness (IMT),diameter and parameters of arterial stiffness [dispensability coefficient (DC),compliance coefficient (CC),stiffness α,stiffness β,pulse wave velocity (PWVβ) ]were measured by ultrasonography with the technology of QIMT and QAS.The thickness of the interventricular septum (IVSd),end-diastolic left ventricular diameter (LVDd) and left ventricular mass (LVM),and parameters of the left ventriculsr function (EF,E/A,E'/A',E/E' and Tei index) were measured by echocardiography.These parameters were compared between two groups.Correlations between the parameters of the arterial stiffness and those of the cardiac function were evaluated by Pearson correlative analysis.Results ①The IVSd,LVM and E/E' ratio were significantly higher in LEAD group than those in control group ( P ＜0.05).There were no significant differences in EF,E/A,E'/A',and Tei index between two groups ( P ＞0.05).②The IMT,α,β,PWVβ of left femoral artery were significantly higher in LEAD group than those in control group,while DC and CC were significantly lower in LEAD group than those in control group ( P ＜0.05).③The E/E' ratio,one of the parameters representing the left ventricular diastolic function,was correlated negatively with CC and positively with α,β,and PWVβ ( P ＜0.05 or P ＜0.01 ).The E'/A' ratio was correlated positively with DC and CC,and negatively with α,β,and PWVβ ( P ＜0.05 or P ＜0.01 ).Both EF and Tei index were not significantly correlated with the above parameters of arterial stiffness ( P ＞0.05).Conclusions Patients with LEAD have thickened femoral IMT,higher arterial stiffness of left femoral artery,as well as impaired left ventricular function.There is a close correlation between the atherosclerosis of the femoral artery and the early left vcntricular dysfunction.

Objective To evaluate the association between the left carotid arterial stiffness and left ventricular diastolic function in patients with lower extremity atherosclerosis (AS).Methods ①A total of 32 patients with AS and 34 control objects were enrolled.The carotid arterial stiffness parameters:compliance coefficient (CC),distensibility coefficient (DC),stiffness parameter (α,β),pulse wave velocity β (PWVβ) were measured by using quality arterial stiffness(QAS) technology.And the values were compared between the two groups.②The parameters of left ventricular (LV) structure and function:LV end-diastolic interventricular septal thickness (IVSd),LV end-diastolic diameter (LVDd),LV end-diastolic wall thickness (PWd),LV ejection fraction (EF),systolic velocity (s'),early-diastolic velocity (e'),Tei index and E/e' ratio were measured by using two-dimensional echocardiography and tissue Doppler.These parameters were compared between the two groups.The association between the carotid arterial stiffness parameters and LV function parameters were analyzed by correlative analysis.Results ①Compared with the control group,the DC and CC were lower,and α,β,PWVβ,IMT were higer than the control group,with statistically significant differences(P ＜0.05).②The IVSd,Tei index and E/e'was significantly higher in the AS group than those in the control group.And the PWd,s',e' were lower than those in the control group (P ＜ 0.05).There was no significant difference in EF between the two groups (P ＞0.05).③The e' was correlated positively with DC and CC (r =0.39,0.36,P ＜0.01),and negatively with α,β,and PWVβ (r =-0.42,-0.42,-0.49,P ＜0.01).Tei index was correlated negatively with DC and CC (r =-0.50,-0.52,P ＜0.01),and positively with α,β,and PWVβ (r =0.58,0.58,0.62,P ＜0.01).The E/e' was correlated regatively with CC (r =-0.27,P ＜0.05),and positively with PWVβ (r =0.28,P ＜0.05).There were no significant correlation between s',EF and the stiffness parameters of carotid artery (P＞0.05).Conclusions In patients with AS,the left carotid artery stiffness increases and left ventricular systolic and diastolic function are impaired.The carotid artery stiffness and left ventricular diastolic function is correlated.Changes in carotid artery stiffness reflect the change in left ventricular diastolic function.