Cancer stands as one of the predominant causes of mortality globally, necessitating ongoing efforts to develop innovative therapeutics. Historically, natural products have been foundational in the quest for anticancer agents. Bulbocodin D (BD) and Bulbocodin C (BC), two bibenzyls derived from Pleione bulbocodioides (Franch.) Rolfe, have demonstrated notable in vitro anticancer activity. In human lung cancer A549 cells, the IC_50s for BD and BC were 11.63 and 11.71 μmol·L^-1, respectively. BD triggered apoptosis, as evidenced by an upsurge in Annexin V-positive cells and elevated protein expression of cleaved-PARP in cancer cells. Furthermore, BD and BC markedly inhibited the migratory and invasive potentials of A549 cells. The altered genes identified through RNA-sequencing analysis were integrated into the CMap dataset, suggesting BD's role as a potential signal transducer and activator of transcription 3 (STAT3) inhibitor. SwissDock and MOE analyses further revealed that both BD and BC exhibited a commendable binding affinity with STAT3. Additionally, a surface plasmon resonance assay confirmed the direct binding affinity between these compounds and STAT3. Notably, treatment with either BD or BC led to a significant reduction in p-STAT3 (Tyr 705) protein levels, regardless of interleukin-6 stimulation in A549 cells. In addition, the extracellular signal-regulated kinase (ERK) was activated after BD or BC treatment. An enhancement in cancer cell mortality was observed upon combined treatment of BD and U0126, the MEK1/2 inhibitor. In conclusion, BD and BC emerge as promising novel STAT3 inhibitors with potential implications in cancer therapy.
Humans ; Lung Neoplasms/metabolism* ; STAT3 Transcription Factor/metabolism* ; Antineoplastic Agents/chemistry* ; A549 Cells ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation

8-hydroxy-2'-deoxyguanosine (8-OHdG) is a sensitive and stable biomarker for evaluating DNA oxidative damage. A rapid and sensitive colloidal gold immunochromatographic strip was developed for 8-OHdG detection by a competitive method. The sample pad (glass cellulose film), bonding pad (glass cellulose film), nitrocellulose film and absorbent pad were pasted on the polyvinyl chloride (PVC) base plate to construct the test strip. Colloidal gold (AuNPs) was prepared by the reduction of chloroauric acid with sodium citrate. 8-OHdG antibody (Ab) was coated on the outer layer of AuNPs to form Ab@AuNPs as a probe. Bovine serum albumin (BSA) and 8-OHdG were conjugated with carbodiimide hydrochloride to prepare an artificial antigen, which was used as the coating antigen of detection line. Goat anti mouse polyclonal antibody IgG was used as the coating antibody of control line. The experimental parameters were optimized including the type of nitrocellulose membrane, the formula of loading solution, and the spraying amount of gold labeled antibody. The results showed that the appropriate nitrocellulose membrane was CN 95. The optimal loading solution included BSA (1%), Tween-20 (3%), sucrose (3%) and NaCl (0.9%). The optimal spraying amount of gold labeled antibody was 4 μL. 8-OHdG can be detected by the strip under visible light, and the level of 8-OHdG in urine can be preliminarily determined by comparing the color intensity of T line and C line. The 8-OHdG concentration in urine was further calculated by the gray value of T line and the threshold of detection was 2.55 μg/L. This colloidal gold immunochromatographic strip is simple, rapid and specific for detecting 8-OHdG in human urine to preliminarily evaluate the human status.
8-Hydroxy-2'-Deoxyguanosine ; Animals ; Antibodies, Monoclonal ; Gold ; Gold Colloid/chemistry* ; Metal Nanoparticles ; Mice ; Sensitivity and Specificity

Objective To explore the value of various diffusion parameters obtained from monoexponential, biexponential and stretched exponential diffusion-weighted imaging models in assessing hepatic fibrosis in chronic hepatitis B(CHB). Methods A total of 52 patients who were diagnosed hepatitis B by the markers of hepatitis and were confirmed by liver biopsy pathology were prospectively recruited between June 2014 and May 2016 in People's Hospital of Henan Province. Concomitantly, 30 healthy volunteers who had no history of hepatitis B and liver dysfunction were enrolled in the control group. All patients underwent multi-b values DWI on a 3.0 T MRI unit. ADC was calculated by using the monoexponential model. True diffusion coefficient(Dt),pseudo diffusion coefficient(Dp)and fraction of perfusion(f)were calculated by using the biexponential model.Distributed diffusion coefficient(DDC)and water molecular diffusion heterogeneity index(α)were calculated by using the stretched exponential model. Liver biopsy specimens were staged according to the degree of hepatic fibrosis (S0 to S4). The Kruskal-Wallis test was employed for the comparison of each parameter among the control group and the fibrosis stage groups. The Mann-Whitney U test was adopted to compare each parameter between fibrosis stage≤S1and≥S2,between≤S2 and≥S3.Spearman rank correlation coefficients were obtained to assess the correlation of the parameters with the fibrosis stages.ROC analysis was used to evaluate the performance of various parameters in predicting stage≥S2 and≥S3.Results The hepatic fibrosis stage distributions were as follows:1 cases with S0,9 cases with S1,22 cases with S2,11 cases with S3,9 cases with S4.ADC,Dt,f and DDC values all showed significant difference among the control group and groups S1,S2,S3,S4(all P<0.05), while Dp and α values showed no significant difference(P>0.05). Dt, DDC and ADC showed a moderate negative correlation with the fibrosis stage(r=-0.630,-0.603 and-0.464,respectively,all P<0.01),and f showed a mild negative correlation with the fibrosis stage(r=-0.379,P<0.05),while Dp and α values showed no correlation with the fibrosis stages(all P>0.05).The ADC, Dt, f and DDC values all showed significant difference between stage≥S2 and≤S1,between stage≥S3 and≤S2(all P<0.05),and the parameter values of the former were greater than those of the latter. While Dp and α values showed no significant difference among both groups(all P>0.05).The AUCs of ADC,Dt,f and DDC values for detecting fibrosis stage ≥S2 were 0.738,0.835,0.740 and 0.831, and the AUCs of ADC, Dt, f and DDC values for detecting fibrosis stage≥S3 were 0.716, 0.811, 0.672 and 0.798. Conclusion The Dt derived from biexponential and DDC derived from stretched exponential DWI could be useful for the staging of hepatic fibrosis in CHB.

Objective To explore the correlation between changes of striatal dopamine D2receptors non-displaceable binding potential (BPND) in 11C-Raclopride PET-CT and brain regional homogeneity (ReHo) derived from resting state functional MRI (fMRI) in patients with first-episode major depressive disorder (MDD) . Methods Patients with first-episode MDD and age and sex matched healthy volunteers accepted brain 11C-Raclopride PET-CT and resting state fMRI. MIAKAT based on MATLAB and Rest 1.8 were used to calculate BPNDof brain dopamine D2receptors and brain ReHo, respectively. Changes of striatal dopamine D2receptors BPNDand brain ReHo values were analyzed by paired-sample t test and two independent sample t-test, and Pearson correlation analysis was used to analyze the correlation between BPNDand ReHo. Results Twenty patients with first-episode MDD were enrolled as MDD group, and 20 healthy volunteers as the control group. The two groups were all right-handed, and there were no statistical differences for age (t =1.33,P=0.19)and gender(χ2=0.10,P=0.75). Compared with the control group, the ReHo in MDD patients increased in the bilateral striatum (caudate nucleus and putamen), bilateral medial prefrontal lobes, and right thalamic (27 to 56 voxels, P＜0.05) and decreased in the left middle frontal gyrus, the left anterior cingulate, the left hippocampal and the right amygdala (21 to 35 voxels, P＜0.05). In addition, compared with the control group, BPNDof bilateral caudate nucleus and putamen dopamine D2receptors in the MDD group were decreased (t=－4.41 to －3.13, P＜0.05). Furthermore, there was a negative correlation between D2 receptors BPNDand ReHo of bilateral caudate nucleus and putamen (r=－0.81 to－0.62, P＜0.05). And there was a negative correlation between ReHo of the bilateral medial prefrontal lobes and BPNDof the same lateral caudate nucleus and putamen D2receptors in the MDD group (r=－0.86 to－0.52, P＜0.05). Besides, ReHo of the left middle frontal gyrus, right thalamic, left anterior cingulate, left hippocampal and right amygdala had no correlation with the D2receptors BPNDof the striatum in the MDD group (－0.27 to 0.39, P＞0.05). Conclusion There were abnormal levels of dopamine neurotransmitter in the cerebral striatum regions and abnormal brain activities in the brain region associated with dopamine reward circuit in the first-episode MDD patients, and there was a correlation between the two abnormalities.

OBJECTIVETo observe the difference in acupuncture for pain threshold at different time points among the groups of 9 TCM constitutions.

METHODSThe cross-sectional survey was adopted to investigate TCM constitutions among 600 subjects and determine 9 TCM constitution types (neutral constitution, qi-deficiency constitution, yang-deficiency constitution, yin-deficiency constitution, phlegm-damp constitution, damp-heat constitution, blood-stagnation constitution, qi-stagnation constitution, special diathesis constitution). The same acupuncture manipulation was applied to Zusanli (ST 36) on the left side in the subjects and the needle was retained for 30 min. The tenderness threshold was detected with 2390 type Von Frey apparatus at different time points, named before acupuncture, at the moment after qi arrival, in 10 min of needle retaining, in 30 min of needle retaining and in 15 min after needle withdrawal in the subjects of 9 TCM constitutions.

RESULTSThe interactive effect happened between the constitution type and time point (P < 0.05). Among the groups of 9 TCM constitutions, the pain threshold values at the moment after qi arrival (except blood-stagnation constitution, qi-stagnation constitution, special diathesis constitution) in 10 min of needle retaining and in 30 min of needle retaining were increased as compared with those before acupuncture separately (P < 0.01), among which, the value increase was the most significant in 30 min of needle retaining. The differences in the pain thresholds were significant in 15 min after needle withdrawal in the groups of neutral constitution and damp-heat constitution as compared with those before acupuncture (both P < 0.01). In 10 min of needle retaining and in 30 min of needle retaining, as compared with the group of neutral constitution, the changes in pain thresholds of the rest abnormal constitutions were apparently lower (all P < 0.05).

CONCLUSIONAcupuncture at Zusanli (ST 36) presents different effects among the groups of different constitution types. The effect maintaining durations are different.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Cross-Sectional Studies ; Female ; Humans ; Male ; Pain Management ; Pain Threshold ; Yang Deficiency ; therapy ; Yin Deficiency ; therapy ; Young Adult

BACKGROUNDThe diagnosis of liver fibrosis is a difficult task at any time using conventional clinical imaging. Intravoxel incoherent motion (IVIM) can be used to investigate both diffusion and perfusion changes in tissues. This study was designed to determine the value of IVIM in the diagnosis and staging of liver fibrosis.

METHODSIVIM examinations were performed on a GE 3.0T MR scanner in 25 patients with liver fibrosis and 25 healthy volunteers as the control group. Patients with liver fibrosis diagnosis were confirmed by pathology and staged on a scale of F0-4. The standard ADC values and the values of a biexponential model (slow ADC (Dslow), fast ADC (Dfast) and fraction of fast ADC (FF)) were measured in three liver regions per person. The mean standard ADC values, Dslow values, Dfast values and FF values from the study group were compared among the right posterior hepatic lobe, right anterior hepatic lobe and medial segment of the left lobe. Receiver Operating Characteristic (ROC) curves and independent-samples t-tests were used to calculate the mean standard ADC values, Dslow values, Dfast values and FF values from the study group and the control group. Spearman rho correlation analysis was used for the stage of liver fibrosis. The liver fibrosis stages between the groups F0-1 and F2-4, the groups F0-2 and F3-4 were compared.

RESULTSAmong the liver fibrosis, there was no significant difference in the mean standard ADC values, Dslow values, Dfast values, and FF values obtained from the right posterior hepatic lobe, right anterior hepatic lobe and medial segment of the left lobe. Using ROC analysis, the Area Under the Curve (AUC) values of standard ADC, Dslow, Dfast, FF were all between 0.7 to 0.9. The mean standard ADC values, Dslow values, Dfast values and FF values of the liver in the study group were significantly lower than the values in the control group (P < 0.05). As the stage of the fibrosis increased, the values decreased by Spearman rho correlation analysis. The mean values (standard ADC, Dslow, Dfast, and FF) of liver fibrosis stages between the groups F0-1 and F2-4, the groups F0-2 and F3-4 showed significant differences (P < 0.05).

CONCLUSIONSIVIM can reflect the conditions of perfusion and diffusion in liver fibrosis and thus distinguish between normal liver and liver fibrosis. The IVIM technique may serve as a valuable tool for detecting and characterizing liver fibrosis, and monitoring its progression in a noninvasive manner.

Adult ; Aged ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Liver ; pathology ; Liver Cirrhosis ; diagnosis ; Male ; Middle Aged

Objective To investigate the outcome of the urethral realignment simultaneous internal fixation in treatment of pelvic fracture associated with posterior urethral disruption. Methods The study included 33 patients with pelvic fracture associated with posterior urethral disruption treated simultaneously by urethral realignment and internal fixation from December 2003 to August 2009. According to AO classification, two patients were with type A2 fracture, six with type B1 ,seven with type B2,five with type B3 ,six with type C1 ,four with type C2 and three with type C3. All the patients were found with complete posterior urethral disruption. Primary urethral realignment combined with emergency open reduction and internal fixation of pelvic fracture were performed in 12 patients. Due to unstable condition, 21 patients underwent primary suprapubic cystostomy and external fixation, sequentially delayed urethral realignment and internal fixation.Results The mean follow-up time of all patients was 37 months ( range, 3-63 months). Of all the patients, 25 patients (76%) regained good without urethral dialation or needed only short term urethral dilatation, and eight patients (24%) suffered from urethral stricture and needed further complex surgery. The incidences of urinary incontinence and erectile dysfunction were 6%(2/33) and 18% (6/33). The erectile dysfunction of two patients was ascribed to sacral nerve injury.At the final follow-up, the mean score was 90.3 points (range, 66-100) according to the Majeed' s scoring system, which showed that the result was excellent in 24 patients, good in eight and fair in one.Conclusions Urethral realignment and simultaneous internal fixation can attain good clinical results for pelvic fracture associated with posterior urethral disruption. It takes advantages of minor surgical trauma,short treatment cycle, good outcome and low complication rate.

ObjectiveTo investigate the results of open reduction with internal fixation for high anterior column fracture of acetabulum with intact true pelvic brim.MethodsFrom January 2006 to January 2010,12 patients suffered high anterior column fracture of acetabulum without involvement of true pelvic brim were identified.There were 9 males and 3 females,with the average of 35.6 years(range,29-46).The injury was caused by crush in 7 cases,smash of heavy object in 3 cases,and fall-down from height in 2 cases.These fractures were classified into three types:isolated high anterior column fracture of acetabulum without involvement of true pelvic brim,high anterior column fracture of acetabulum without involvement of true pelvic brim with posterior wall fracture of acetabulum and commminuted high anterior column fracture of acetabulum without involvement of true pelvic brim according to present and unpresent of posterior wall fracture.Five cases suffered isolated fracture and 2 cases associated smaller and nondisplacement fracture fragment of posterior wall were reduced and fixed by buttress plate of iliac crest and lag screw of anterior border of ilium through an iliofemoral approach; 4 cases with displacement posterior wall fracture of acetabulum were reduced and reconstructed by buttress plate and lag screws of through combinations of anterior and posterior approaches.One case suffered comminuted fracture were reduced and reconstructed by plates and screws of through extended iliofemoral approach.ResultsThe mean follow-up time of all patients was 26.7 months (range,14-37months).The quality of reduction was grade as anatomical in 8 patients,imperfect in 3,poor in 1 by Matta's score system.The fracture union was uneventful.There was nonunion and loss of internal fixation.At the final follow-up,the mean score was 16.8 (range,11-18),7 cases were graded as excellent,4 good,1 fair,according to modified Merle d'Aubigne and Postel score system.Heterotopic ossification and traumatic osteoarthritis were recorded in 1 patient.ConclusionGood clinical results can be achieved by anatomical reduction and rigid fixation through optimal approach.

Bone turnover is regulated by local concentrations of cytokines such as osteoprotegerin (OPG) and receptor activator of nuclear factor kappaB ligand (RANKL). To explore the in vivo biological function of Opg and the mechanism of osteoporosis due to deficiency of Opg, Opg knockout mice have been generated through homologous recombination. Opg-/- mice exhibit a sharply decrease in bone density and strength as expected. The number of osteoclasts in Opg-/- mice significantly increases. Morphologically, osteoclasts appear more cuboidal in shape in Opg-/- mice than those of wt mice, suggesting that active osteoclastogenesis occurs in the absence of Opg. In consistent with this finding, an increase of osteoblast activity was also observed with accelerated mineral accumulation rate by histomorphometric measurement and elevated serum alkaline phosphatase activity (ALP) in Opg-/- mice. Interestingly, more than 50% of 2-month-old Opg-/- mice manifest medial calcification of aorta with comparable serum concentrations of calcium and phosphorus to wt mice. In conclusion, Opg-/- mice have a high-bone-rurnover type osteoporosis. The aortic calcification in Opg-/- mice is not due to abnormality of calcium and phosphorus metabolism. The mechanism underlying aortic calcification in Opg-/- mice needs to be further investigated.

Adiponectin is an adipocyte-derived secretory protein. It was found to be associated with insulin resistance, inflammation and arteriosclerosis. To further study the biological function and expression of adiponection in vivo, adipoenctin gene knock-out and LacZ gene knock-in mouse model was constructed. Gene targeting strategy was designed to replace part of exon 2 and exon 3 of adiponectin gene with full length LacZ gene in frame with remaining upstream ATG and signal peptide sequence of exon 2. The targeting vector (Adipo-LacZ-XpPNT) was constructed and verified by restriction enzyme digestion and sequencing. CJ7 ES cells were transfected with targeting vector linearized by NotⅠ digestion, selected in the medium containing both G418 and ganciclovoir. Resistant clones were screened by PCR and further confirmed by Southern blot for correct homologous recombinants. Chimera mice were obtained by routing microinjection of homologous recombined ES cells into blastocysts. After mating, mice heterozygous and further homozygous for adiponectin knockout and LacZ gene knock-in were established. Expression of both endogenous adiponectin and exogenous LacZ gene in mouse tissues and sera were detected by RT-PCR, Northern-blot, Western blot and ELISA. The results show that adiponectin was disrupted at both mRNA and protein levels. LacZ gene is expressed exclusively in adipose tissue of mutant mice. Its expression profile is identical to endogenous adiponection. Unexpectedly, LacZ activity could not be detected in both adipose tissue and serum although LacZ protein can be detected in adipose tissue but not in serum of mutant mice. In conclusion, mice homozygous for adiponectin knockout and LacZ gene knock-in have been successfully constructed. Mutant mice display LacZ expression profile identical to endogenous adiponectin albeit neither LacZ activity nor protein can be detected in serum of mutant mice.