Objective To evaluate the effect of Wnt5A gene overexpression on cytoskeletal proteins of melanocytes after the plasmid containing the Wnt5A gene is transfected into primary melanocytes.Methods In vitro cultured primary human melanocytes were divided into three groups:blank control group receiving no treatment,negative control group transfected with endotoxin-free pcDNA3.1 (+)empty vector by Lipo3000 in Opti-MEM medium,Wnt5A plasnid group transfected with endotoxin-free pcDNA3.1 (+) vector containing the Wnt5A gene by Lipo3000 in Opti-MEM medium.After the transfection,quantitative PCR (qPCR) was performed to measure the mRNA expression of Wnt5A,ras-related C3 botulinum toxin substrate 1 (Rac1),filamentous actin (F-actin) and β-tubulin,Western blot analysis to determine the protein expression of Wnt5A,receptor tyrosine kinase like orphan receptor 2 (ROR2),Rac1,F-actin and β-tubulin,and an immunofluorescence assay (IFA) to observe the expression of cytoskeletal proteins.Results qPCR showed significant differences in the mRNA expression of the Wnt5A gene and its downstream genes Rac1 and F-actin among the Wnt5A plasmid group,negative control group and blank control group (F =1374.179,112.576,66.458,respectively,all P ＜ 0.01),but there was no significant difference in the mRNA expression of β-tubulin among the three groups (P ＞ 0.05).Additionally,the Wnt5A plasmid group showed significantly higher mRNA expression of Wnt5A,Rac1 and F-actin compared with the blank control group and negative control group (all P ＜ 0.05).As Western blot analysis revealed,compared with the blank control group and negative control group,the Wnt5A plasmid group showed significantly higher Wnt5A protein expression (both P ＜ 0.05),but significantly lower protein expression of Rac 1,ROR2 and F-actin (all P ＜ 0.05).However,no significant difference in β-tubulin protein expression was observed among the three groups (P ＞ 0.05).IFA showed no obvious difference in the fluorescence intensity of β-tubulin or F-actin between the Wnt5A group and the two control groups,but melanocytes showed larger size and increased number of dendrites,and the cytoskeleton changed dramatically with varying fluorescence intensity of F-actin,fuzzy texture,fractured or locally clustered tonofilaments in the Wnt5A group.Conclusion The overexpression of the Wnt5A gene in melanocytes can regulate the mRNA and protein expression of cytoskeletal proteins,nake melanocytes larger and more dendritic,and cause changes in the cytoskeleton,which may facilitate the transportation of melanosomes,and participate in the occurrence of hyperpigmented diseases.

Objective To discuss the cause and management of delayed hemorrhage after minimally invasive percutaneous nephrolithotomy (PCNL). Methods From 2004 to 2009,there were 13 cases (10 men and 3 women, mean age 47 years) developed severe bleeding following PCNL. The cause of hemorrhage and treatments were retrospectively analyzed and summarized. Results The time of hemorrhage was 5-40 d. In 7 of 13 cases, the bleeding were was controlled by complete bed rest,hemostatic,balloon compression and clamped nephrostomy tube. Six of 13 underwent selective renal arteriography after ineffective with conservative treatment. The DSA showed 4 were false aneurysm and 2 were arteriovenous fistula. All the 6 cases were treated by the super-selective arteriolar embolization. The hematuria was disappeared 1- 3 d later. The intravenous pyelogram revealed the renal function kept well during follow-up visit. Conclusions Delayed hemorrhage is one of the severe complications, which may be caused when the renal puncture passage established as forming false aneurysm and arteriovenous fistula. The renal arteriography and super-selective arteriolar embolization could be a safe and effective treatment for the severe hemorrhage after PCNL.

Objective:To observe the natural course of cerebral contusion and laceration combined with hematoma formation and analyze the risk factors for its progression.Methods:Patients with cerebral contusion and laceration combined with hematoma formation admitted to our hospital from September 2017 to March 2020 were prospectively selected; and they were divided into progressive and non-progressive groups according to progression of cerebral contusion and laceration combined with hematoma formation. The clinical data of the two groups of patients were compared, and multivariate Logistic regression was used to analyze the independent influencing factors for progressive cerebral contusion and laceration combined with hematoma formation.Results:A total of 197 patients with cerebral contusion and laceration combined with hematoma formation were included in this study, of which, 61 were treated with craniotomy and 136 were treated conservatively; 85 patients had progressive cerebral contusion and laceration combined with hematoma formation and 112 patients had non-progressive cerebral contusion and laceration combined with hematoma formation. As compared with those in the non-progressive group, the baseline Glasgow Coma Scale (GCS) scores of the progressive group were lower, hematoma volume by second CT scan was larger, distance from the center of cerebral contusion and laceration or hematoma to the nearest cortex was shorter, platelet count and thrombin time increased, fibrinogen (FIB) content decreased, and proportion of patients with multiple lesions in the first CT scan was higher in the progressive group, with significant differences ( P<0.05). Multivariate Logistic regression analysis showed that the distance from the center of cerebral contusion and laceration or hematoma to the nearest cortex<1 cm, plasma FIB<2 g/L, multiple lesions of cerebral contusion and laceration or hematoma on first CT scan were risk factors for progression in patients with cerebral contusion and laceration combined with hematoma formation ( OR=6.654, 95%CI: 1.391-35.089, P=0.025; OR=5.617, 95%CI: 1.136-28.022, P=0.034; OR=4.629, 95%CI: 1.178-20.071, P=0.031). Conclusion:The patients with short distance from the center of cerebral contusion and laceration or hematoma to the nearest cortex, low plasma FIB, and multiple lesions of cerebral contusion and laceration or hematoma on first CT scan are prone to have progressive cerebral contusion and hematoma formation.