The p21-activated kinase 1 (PAK1) is a member of the P21-activated protein kinase family that plays an important role in the proliferation and on cogenesis of pancreatic cancer. PAK1 is an important target for the treatment of pancreatic cancer. At present, akinase inhibitor targeting PAK1 is still in the preclinical research stage. Therefore, screening for new PAK1 kinase inhibitors is of great significance. In this study the natural compound celastrol was found to have a significant inhibitory effect on PAK1, with an IC₅₀ value of 3.614 μmol·L^-1. Molecular docking results showed that celastrol had good binding to PAK1. An MTT assay indicated that celastrol inhibited the proliferation of pancreatic cancer cells BxPC-3 and PANC-1. Mechanistic studies revealed that the inhibition of pancreatic cancer cells by celastrol was reversed by PAK1 siRNA. Celastrol inhibited PAK1 and the subsequent activation of downstream signaling pathways, thereby activating apoptosis signaling pathways and triggering apoptosis in pancreatic cancer cells. These findings suggested that celastrol induced apoptosis in pancreatic cancer cells by suppressing the PAK1 kinase signaling pathway and has potential value for the treatment of pancreatic cancer.

OBJECTIVE: To investigate the mutation of RUNX1 gene in patients with myelodysplastic syndrome (MDS) and its correlation with other gene mutations and some clinical parameters. METHODS: The mutations of RUNX1, DNMT3A, TET2, IDH1/2, NPM1, FLT3-ITD and C-KIT in 170 patients with MDS were detected by direct and indirect sequencing of genomic DNA-PCR amplification products. RESULTS: The RUNX1 mutation was found in 23 patients (13.5 %, 23/170). Among the 170 patients, other most frequent mutation was TET2 (11.2%, 19/170), followed by mutations in DNMT3A (9.4%, 16/170), NPM1 (8.2%, 14/170), IDH2 (4.1%, 7/170)、FLT3-ITD (2.9%, 5/170), IDH1 (1.7%, 3/170) and c-KIT (0.58%, 1/170). The most common coexisting mutations were TET2 (5/23). The RUNX1-mutated group showed significantly higher leukocyte levels, higher percentages of blast cells, higher incidences of leukemia transformation and lower platelet counts in comparison with RUNX1 non-mutation group (P＜0.05). whereas there were no statistically significant difference in age, MDS subtype, karyotype and hemoglobin level between 2 groups (P＞0.05). Seventeen patients harboring RUNX1 mutations were followed up and almost 47.05% (8/17) of the patients progressed into acute myeloid leukemia (AML). The rates of transformation into AML in ASXL1-mutation group was significantly higher than that in ASXLL- non-mutation group (47.05% vs 11.7%) (P=0.001). CONCLUSION The incidence of RUNX1 mutation is high in MDS patients. The RUNX1-mutated patients have higher leukocyte level, higher percentages of blast cells, higher incidences of leukemia transformation and lower platelet count.

OBJECTIVE: To discuss the long-term outcome of convex epiphysiodesis in the treatment for congenital scoliosis (CS). METHODS: The clinical data of 22 patients with hemivertebral deformity undergoing convex epiphysiodesis from the October 1998 to Febuary 2008 were respectively analyzed. There were 12 males and 10 females. The whole spine anteroposterior radiographs were taken preoperatively, at 3-month postoperatively and at the final follow-up to measure the main curve and the compensatory curve. The progression rate was calculated for each patient. Observing the correlation between the progression rate and annual progression of the scoliosis and age, gender, hemivertebral number, hemivertebral position, preoperative main curve Cobb angle and compensatory curve Cobb angle, comparing different ages, genders, hemivertebral number and position, and preoperative main curve Cobb angle on the progression of postoperative curve. RESULTS: The mean Cobb angle of main curve changed from (40.5±9.8) ° before surgery to (39.5±11.1) ° at 3 months after surgery, which significantly increased to (46.8±13.9) ° in the final follow-up. Meanwhile the mean Cobb angle of compensatory curve was changed from (20.1±10.8) ° before surgery to (23.0±11.1) °, which significantly increased to (29.9±11.5) ° in the final follow-up. There were no significant differences in the Cobb angle of the main curve and the compensatory curve between postoperative 3 months and before operation (>0.05). The difference between the final follow-up and the preoperative, postoperative 3 months was statistically significant (<0.01). Twenty patients experienced progression of both main curve and compensatory curve, with a mean progression rate of (19.2±17.9)% for main curve and (39.6±37.0)% for compensatory curve. The annual progression volume was (1.5± 1.4) ° for main curve and (1.4±1.3) ° for compensatory curve. Three patients underwent lateral convex orthopedic internal fixation due to postoperative scoliosis progression. The curve progression was significantly correlated with age at the time of surgery and hemivertebral number. There was a significant correlation between the age of the operation, the main curve angle, the preoperative compensatory curve angle and the annual progression volume of the main curve (<0.05). CONCLUSION The convex epiphysiodesis technique cannot effectively prevent curve progression of CS patients in the long-term follow-up. It is not recommended to apply this technique to the treatment of patients with congenital hemivertebrae.

Objective: To evaluate efficacy and safety of retrograde intrarenal stone surgery for treatment of upper urinary calculi. Methods: The clinical data of 640 patients with upper urinary tract calculi treated by retrograde intrarenal stone surgery (RIRS) in Renmin Hospital of Wuhan University from April 2016 to January 2019 were retrospectively analyzed. There were 424 males and 216 females. The awerage age was (46.2±12.8) years old, ranging 18 to 76 years old. The maximum diameter of the stone is (1.4±0.7) cm, ranging 0.6-3.2 cm. There were 126 cases with inferior calculi and 514 cases with non-lurgical calculi. There were 196 cases with unilateral ureteral calculi, 118 unilateral ureteral calculi cases with renal calculi, 236 cases with unilateral renal stones, and 90 cases with double kidney stones. 104 cases were placed with double J tube before operation and 496 cases were not placed before operation. There were 8 cases of horseshoe kidney, 30 cases of isolated kidney with renal insufficiency, 4 cases of pelvic ectopic kidney with dysplasia, 6 cases of congenital ureteral malformation and 2 cases of sponge kidney. Preoperative average hemoglobin was (133.2±5.6)g/L, ranging 126-188 g/L.And average serum creatinine was (84.4±12.2)μmol/L, ranging 74-242μmol/L before operation. All patients were treated with general anesthesia under the lithotomy position. The ureteroscopy combined with holmium laser lithotripsy was performed.The 200μm fiber was used, which the parameters were set as 12-45 W(0.5-1.5 J/10-30 Hz). The stone baskets were used to take stones according to actual conditions. The operation was performed by doctors of the same qualifications. Results: All patients underwent successful operation. The mean operation time was (45.6±14.6)min. The average postoperative hospitalization was (4.8±1.5)d. The postoperative serious complication rate was 0.9%, including(2 cases of sepsis and 1 case of subcapsular hematoma. Of the 640 patients, 596 were admitted to the hospital for a double J tube and 44 were lost of follow-up. 552 patients met the stone removal criteria, 44 patients did not meet the stone removal criteria for other treatments, such as extracorporeal shock wave lithotripsy, ureteroscopy or observed regularly. The stone-free rate (SFR) was 92.6% (552/596) after 1-3 months. On the first postoperative day, serum creatinine was (76.0±10.6)(58-156) μmol/L, and postoperative hemoglobin was (126.4±9. 6)(120-176) g/L. There was no significant difference in preoperative and postoperative hemoglobin (t=2.02, P=0.064). Preoperative and postoperative creatinine (t=64.76, P<0.05) was statistically significant. Meanwhile, the stone size (χ²=29.569, P<0.05) and position (χ²=44.949, P<0.05) versus SFR the impact was statistically significant. Multivariate regression analysis showed that stone size was not an independent risk factor for stone clearance (P=0.639). The stone position was an independent risk factor for stone clearance (P=0.013). Conclusions RIRS is a reliable treatment for small and medium calculi patients of the upper urinary tract. The curative effect of stone removal is clear, the complications are few, the safety is high. However, there are certain limitations to the efficacy in the treatment of large stones and lower calculi. Lower calculi is the independent risk factor for the treatment of efficacy.

Objective: To evaluate diagnostic performance of Todd-Hewitt (T-H) broth culture method, direct culture method, liquid chromogenic culture method, and loop-mediated isothermal amplification (LAMP) method for screening group B streptococcus (GBS) during late pregnancy. Methods: In the retrospective study, the rectal vaginal secretions samples were collected from pregnant women at 35 to 37 weeks at the obstetrics clinic of Guangzhou Women and Children′s Medical Center affiliated to Guangzhou Medical University during October 2016 to April 2018. For the purposes of clinical evaluation, T-H broth culture was used as the standard reference method, and double-blind trials were used to evaluate diagnostic performance of direct culture method, liquid chromogenic culture method, and LAMP method for screening group B streptococcus during late pregnancy in three research stages. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), coincidence rate and Yoden index for each method were calculated. Also, the level of agreement between each method and T-H broth was assessed using the kappa (k) coefficient. Results: A total of 969 specimens were detected by the T-H enrichment culture method, and 90 were positive (9.3%). The sensitivities from high to low were LAMP method [100% (25/25)], direct culture method [81.5% (22/27), 95%CI:65.8%-97.1%], and liquid color culture method [71.1% (27/38), 95%CI:55.9%-86.2%]. Specificities were direct culture method [100% (282/282)], liquid color culture method [98.1% (455/464), 95%CI:96.8%-99.3%], and LAMP method [94.0% (125/133), 95%CI: 89.9%-98.1%]. The coincidence rates were direct culture method [98.4% (22+282)/309], liquid color culture method [96.0% (27+455)/502], and LAMP method [94.9% (25+125)/158]. The Kappa values of the direct culture method (0.889), LAMP method (0.832) and the enrichment culture method were all ≥0.75, and that of the liquid color culture method was 0.708. The false negative rate of direct culture method was 18.5% (5/27), and no false negative case by LAMP method, but its false positive rate was 6.0% (8/133). The false negative rate and false positive rate of liquid color culture method were 28.9% (11/38) and 1.9% (9/464), respectively. Conclusions Of the three screening methods compared in this study, only the LAMP method has the advantages in sensitivity, specificity, and coincidence rate compared with T-H enriched culture method, while the others have a certain degree of false negatives rate. The clinical laboratory can introduce these methods based on laboratory facilities and staffing, or refer to the European and American guidelines and combine the recommended antenatal GBS screening method with intrapartum nucleic acid amplification tests to best meet the clinical demands.

Objective: To investigate the efficacy of alternative donor (AD) in the treatment of aplastic anemia (AA) in children. Methods: The clinical data of AA children who received AD HSCT in our center from Apr. 2010 to Dec. 2016 were retrospectively analyzed. The overall survival (OS) rate, implant success rate, incidence of acute and chronic graft-versus-host disease (GVHD) were statistically analyzed. Results: A total of 109 children with acquired AA, including 64 severe AA (SAA) , 32 very severe AA (VSAA) and 13 transfusion dependent non-severe AA (NSAA) , were recruited in this retrospective AD HSCT study, the median age was 6 (0.8-18) years old. Of them, 44 patients with 10/10 matched unrelated donor (MUD) , 44 patients with mismatched unrelated donor (MMUD) and 21 patients with mismatched related donor (MMRD) . All patients did not receive ATG before HSCT and the active infection was excluded. Except 3 patients suffered from a second graft failure (2 of them rescued by second HSCT) , 106/109 (97.2%) were engrafted with neutrophil and platelet recovery occurring at a median of 13 days (range, 9-19) and 16 days (range, 10-81) post-transplant. Until day 100 post transplantation, the incidence was 74.3% (81/109) for acute GVHD (aGVHD) and 39.4% (43/109) for grade Ⅱ-Ⅳ aGVHD, 30.7% (31/101) and 9.9% (10/101) for overall chronic GVHD (cGVHD) and moderate cGVHD, respectively, and nobody developed an extend cGVHD. After median follow up of 39 (0.7-103) months for all patients, 13 of 109 patients died. The estimated 5-year overall survival (OS) of the entire cohort was 88.1% (95%CI 81.1%-91.4%) with no difference among the MUD, MMUD and MMRD cohort (93.2%, 84.1% and 85.7%, respectively, P=0.361) . Conclusion These excellent outcomes suggest that unmanipulated AD PBSC is a good HSCT source for children with SAA. It’s reasonable to consider AD HSCT as first line therapy for SAA children without matched sibling donor. Better strategies are required to prevent GVHD.

OBJECTIVE Attempt to visualize the hypopharyngeal anatomy and related structures in normal adults with ultrasound. METHODS Sonographic features of laryngopharynx were confirmed by comparable tissue sections of 6 corpses at the same level. Then, the laryngopharynx of 30 men and 30 women were examined by ult rasound and thei r sonographic images were evaluated. RESULTS In male or female volunteers, perfect visualization pattern was obtained in all (100％) pyriform sinuses, 60％ or 93.3％ postcricoid areas,66.7％ or 93.3％ posterior pharyngeal walls, 96.7％ or 100％ thyroid cartilages, 63.3％ or 93.3％ arytenoid cartilages, 70％ or 93.3％ vocal cords,and 83.3％ or 96.7％ ventricular bands. CONCLUSION The ultrasonic visualization of laryngopharynx and its related structures are satisfactory in healthy adult women. For adult men, the pyriform sinus and thyroid cartilage can be best identified while the identification of vocal cords and ventricular bands is less satisfactory. However, postcricoid region, pharyngeal posterior wall, and arytenoid cartilage are the most difficult to detect.

Objective To evaluate the clinical curative effect of multiple interventional techniques with skilled manipulation in treating common bile duct stones. Methods The clinical data of 36 patients with common bile duct stones, who were treated with percutaneous transhepatic puncture of intrahepatic bile duct and multiple interventional techniques, were retrospectively analyzed. Before operation, according to CT or MRI findings the optimal puncture path was selected to perform cholangiography so as to reconfirm the position, number and size of the stones. Depending on the size of the stone, the corresponding sized lithotripsy basket and balloon were selected. The duodenal papilla was dilated by balloon, and the stone was pushed into the duodenum with a balloon. After complete removal of stones, the internal and external biliary drainage tubes were routinely implanted and remained there for two weeks. Results The operation was successfully accomplished in 35 patients, the success rate was 97.2％. Successful removal of stones with single procedure was accomplished in 32 patients, and in 3 patients the complete removal of stones was achieved by secondary procedure. No serious complications were observed during the operation. Early postoperative complications included hyperamylasemia (n=2), bile peritonitis (n=2), small amount of bloody bile (n=3) . No biliary perforation occurred. Conclusion In treating common bile duct stones, the combination use of multiple interventional techniques with skilled manipulation has high success rate, low risk and less complications, therefore, it is an effective therapeutic method

AIM:To observe the expression of hypoxia-inducible factor-1α (HIF-1α) , vascular endothelial growth factor A (VEGFA) and vascular endothelial cadherin (VE-cadherin) in cultured rat renal glomerular endothelial cells (rRGECs) exposed to glucose at different concentrations in vitro, and to verify the hypothesis that salidroside attenuates high glucose (HG) -induced injury of rRGECs by boosting HIF-1αlevel.METHODS:rRGECs were divided into 4group:normal glucose (NG) group, HG groups, hypertonic group and salidroside+HG group.The viability of rRGECs was measured by MTT assay.The mRNA expression of VEGFA, VE-cadherin and HIF-1αwas detected by RT-qPCR.The protein expression of HIF-1αwas determined by Western blot.RESULTS:Compared with NG group, the mRNA and protein expression of HIF-1αwas increased when the rRGECs were treated with glucose at concentration of 20 mmol/L for 24h (P<0.01).Compared with NG group, the mRNA expression of HIF-1αwas decreased in HG groups for 120 h (P<0.05).Compared with NG group, the mRNA expression of VE-cadherin was significantly down-regulated in HG groups for24 h or 120 h (P<0.05).Compared with NG group, the mRNA expression of VEGFA was increased in HG groups at 24h (P<0.05) , while the mRNA expression of VEGFA was decreased at 120 h (P<0.05).Compared with NG group, no statistical difference in the mRNA expression levels of HIF-1α, VE-cadherin and VEGFA in DM group was observed.Compared with HG group, salidroside promoted the viability of rRGECs (P<0.01) , and up-regulated the mRNA expression of HIF-1αand VE-cadherin, and the protein expression of HIF-1α (P<0.05).CONCLUSION:High glucose regulates HIF-1αexpression in rRGECs in connection with cell viability, the concentration of glucose, the culture time and HIF/VEGF signaling.Salidroside promotes rRGEC growth against high glucose-induced cell apoptosis via up-regulating the expression of HIF-1α.

AIM:To investigate whether ellagic acid (EA) attenuates hypoxic-ischemic encephalopathy (HIE) by down-regulating autophagy.METHODS:In vivo, Sprague-Dawley rats (n=17) were randomly divided into 3groups:5 rats for sham group, 6 rats for HIE group and 6 rats for HIE+EA pretreatment group.The rats in HIE+EA pretreatment group were treated with EA (10 mg/kg, 10 m L/kg, suspended in corn oil, ig).After 24 h of operation, the rats from each group were sacrificed and their brains were collected.TTC staining and HE staining were used to define the infarct areas and brain structure.The autophagy-related proteins beclin-1, P62, LC3-II/-I and Atg5 in the cortex in each group were compared by Western blot.In vitro, PC12 cells were divided into 3 groups:control group, Coand CoEA pretreatment group.Co800μmol/L was added to the PC12 cells to induce an anoxic environment.The PC12 cells were pretreated with EA at 8μmol/L and the cell viability was measured by CCK-8 assay.The production of reactive oxidative species (ROS) in the cells was detected by flow cytometry with DCFH-DA staining.MDC staining and TM-RE staining were applied to reflect the extent of autophagy and the state of apoptosis, respectively.The autophagy-related proteins in PC12 cells were also investigated.RESULTS:In HIE group, 7-day-old rats were given the operations and the their large infarct areas in the hemisphere were observed by TTC staining.HE staining displayed the injured hemispheres which contained few neurons, and exhibited edema status and serious structural damage.EA pretreatment decreased the infarct area and alleviated the damage to hemisphere with more visible neurons, compared with HIE group.Compared with sham group, the levels of autophagy-related proteins Atg5, beclin-1 and LC3-II/-I in the cortex were increased (P<0.01) , and P62 protein expression was decreased (P<0.01) in HIE group.Compared with HIE group, the protein expression of Atg5, beclin-1 and LC3-II/-I was decreased (P<0.01) and P62 protein expression was increased in HIE+EA pretreatment group (P<0.01).In vitro, compared with CoPC12 cells in CoEA pretreatment group showed a lower ROS level.Moreover, the cells in CoEA pretreatment group exhibited higher mitochondrial membrane potential than that in CoMDC staining in Coshowed high value of fluorescence and increased number of autophagosomes.EA pretreatment reduced the number of autophagosomes and the extent of autophagy to protect PC12cells.Furthermore, the protein levels of Atg5, beclin-1 and LC3-II/-I in Cowere higher (P<0.01) , and the protein expression of P62 was lower (P<0.01) than those in control group.In CoEA pretreatment group, the protein levels of Atg5, beclin-1 and LC3-II/-I were decreased (P<0.01) and the protein expression of P62 was increased as compared with Co (P<0.01).CONCLUSION:EA pretreatment attenuates autophagy to protect the neurons against HIE injury.