Objective To investigate the method and effectiveness of wide local excision combined with Mohs micrographic surgery for dermatofibrosarcoma protuberans (DFSP). Methods Between January 2007 and January 2010, 17 patients with DFSP were treated. There were 9 males and 8 females with an average age of 33.2 years (range, 16-55 years). Thelesions were located at head and neck (2 cases), trunk (12 cases), extremity (2 cases), and perineal region (1 case). There were 6 cases of primary DFSP and 11 cases of relapsed DFSP. The lesions presented as single or multitude nodules or fusion nodules with skin withering, scar, en plaque in the center and with ill-defined margins. The diameter of lesions ranged from 0.8 to 9.7 cm (mean, 4.3 cm). No distant metastasis or lymphatic metastasis occurred in all cases. After tumors resection by wide local excision combined with Mohs micrographic surgery, the wounds were repaired by direct suture in 3 patients, skin grafting in 9 patients, and local skin flap in 5 patients. Results Wide local excision and Mohs micrographic surgery were carried out once in 13 patients, twice in 3 patients, and three times in 1 patient with an average operation time of 98.6 minutes (range, 56-219 minutes). Primary heal ing of wound and donor site were achieved with no necrosis of skin grafting and skin flap. All patients were followed up 8-34 months (mean, 21.7 months) with no recurrence. Conclusion Wide local excision combined with Mohs micrographic surgery could treat DFSP, which has the advantages of shorter operation time, radical resection, and less injury.
【Abstract】 Objective To explore the optimal dosage, timing and cytotoxicity of bromodeoxyuridine (BrdU) labelling for rabbit adipose-derived stromal stem cells (ADSCs) in vitro so as to confirm its feasibil ity for stem cells labell ing and tracer means. Methods Six rabbits were used in this experiment, aged 8-12 weeks, weighing 1.5-2.0 kg and neglecting their gender. 1-2 mL fat was removed, the ADSCs were isolated and cultured using the adherence method in vitro . The 3rd passage of ADSCs was incubated with BrdU at 5, 10, 15 and 20 μg/mL (groups A, B, C and D）for 12, 24, 48 and 72 hours to identify the optimal BrdU concentration and incubating time for cell labell ing. Immunohistochemistry and trypanblau strain were performed respectively to calculate the labell ing index (positive rate) and the cells’ activity for different time after BrdU labell ing. The ADSCs without BrdU labell ing were used as control (Group E）. Results The main appearance of primary ADSCs was short fusiform shape, and of the 3rd passage ADSCs long fusiform shape. The 3rd passage of ADSCs could differentiate into osteoblastsand adipocytes under corresponding inductive medium. The ADSCs’ nucleus show green fluor under fluorescence microscope after labeled by the BrdU. The labell ing ratio increased in groups A, B, C and D after incubating 12 hours, the mean labell ing ratio were 30.6％ ±2.3％，32.4％ ±1.9％，45.8％ ±1.8％，50.8％ ±3.1％ , respectively, and the labell ing ratio of Group E was 0. There were significant differences between groups C, D and Group A (P lt; 0.01). The labell ing ratio of groups A, B, C and D were 45.9％ ±2.0％，87.9％ ±3.3％，90.6％ ±2.9％，91.7％ ±3.2％，respectively after 24 hours and the labell ing ratio of Group E was 0. There were significant differences between groups B, C, D and Group A (P lt; 0.01). The results of all groups after incubating48 hours and 72 h ours were similar to that after incubating 24 hours. The cell counting of groups A, B, C and D were better than that of Group E, but showing no siginificant differences（P gt; 0.05）. Conclusion The most appropriate time for BrdU labell ing ADSCs is 48 hours, the most appropriate concentration is 10 μg/mL. The labell ing rate is high and cytotoxicity is l ittle.
Objective To evaluate hepatic functional reserve and investigate the clinical value through measuring hepatic functional blood flow by D-sorbitol clearance rate and liver volume changes with CT. Methods Ninety-two patients with portal hypertension due to posthepatic cirrhosis were investigated (cirrhosis group). Twenty healthy volunteers were used as control group. D-sorbitol was infused intravenously at a steady rate. Blood and urine were collected and recorded once before infusion and at 120, 150 and 180 min after infusion, and their concentrations of D-sorbitol were examined by enzyme spectrophotometry. From pharmacokinetic equations, hepatic clearance rate of D-sorbitol (CLH) was calculated. Total hepatic blood flow (QTOTAL) was measured by Doppler sonography, intrahepatic shunt rate (RINS) was obtained. The liver volume change rate was obtained in patients with cirrhosis through the abdominal CT scan. The relations among the indicators, Child classification and postoperative complications were studied. Results After D-sorbitol was infused intravenously for 120 min, the plasma concentration was at the steady state. The plasma concentration was (0.189±0.05) mmol/L in control group and (0.358±0.06) mmol/L in cirrhosis group (Plt;0.01). CLH was (1 248.3±210.5) ml/min in control group and (812.7±112.4) ml/min in cirrhosis group (Plt;0.01). Although QTOTAL in cirrhosis group was declined, compared with the control group 〔(1 280.6±131.4) ml/min vs. (1 362.4±126.9) ml/min〕, Pgt;0.05, while RINS increased markedly 〔(36.54±10.65)% vs. (8.37±3.32)%, Plt;0.01〕. In cirrhosis group, the mean liver volume of Child A, B and C patients were (1 057±249) cm3, (851±148) cm3 and (663±77) cm3 respectively. There were significant differences among the mean liver volume (Plt;0.05). The liver volume was significantly smaller in Child B and C patients than that in Child A (Plt;0.05, Plt;0.01). When CLH was less than 600 ml/min, and liver volume decreased by more than 40%, postoperative complications increased significantly. CLH and the liver volume change rate were not in absolutely good accordance with Child classification. Conclusion The hepatic clearance of D-sorbitol and the quantitative determination of the liver volume with CT can be an objective evaluation of the liver metabolism of the inherent capacity and the hepatic functional blood flow changes. It contributes to the correct understanding of the hepatic functional reserve and lay the foundation for determining a reasonable treatment plan, surgical methods and time.
As most patients of central serous retinopathy (CSC), the symptoms of acute onset will alleviate by oneself after 4-6 months. About 30%-50% of patients with CSC experience chronic or recurrent cases. Resulting in persistent neurosensory detachments and subretinal fluid, causing significant vision loss. Mineralocorticoid receptor (MR) is a kind of nuclear hormone receptors, plays a role in theregulation of water and electrolyte balance. Excessive MR signaling is associated with many diseases. Study found that MR antagonists decreased the thickness of the retina and improved in vision, there was no serious adverse reactions during the period of treatment for chronic CSC. Initial dose of MR antagonists was 25 mg per day, 1 week later, dosage was increased to 50 mg per day, and treatment for about 3 months. There is no conclusive effective treatment and the dosage are still unknown. MR antagonists may be a safe and effective way to treat chronic CSC, though evidence is scant. Prospective, multicenter, large-scale trials is required.
Objective To formulate an individualized evidence-based treatment for a first-diagnosed patient with coronary artery-pulmonary artery fistula. Methods Aiming at the issue of whether interventional operation was necessary for first-diagnosed coronary artery-pulmonary artery fistula or not, the computer retrieval was conducted in the US National Guideline Clearinghouse, The Cochrane Library, PubMed and MEDLINE from 1990 to 2011, to collect and assess the best evidence of relevant systematic reviews, randomized controlled trials, controlled clinical trials and treatment guidelines, in order to be applied in clinical treatment. Results There were 1 clinical guideline for treating coronary artery fistula and 3 different high-quality evidence studies were retrieved. The results showed percutaneous coronary intervention was the best treatment currently. According to the obtained evidence and patient’s willingness, the relevant examinations were taken, and the preoperative preparation for percutaneous coronary intervention was done actively after the patient was admitted. Three days after hospitalization, the selective coronary angiography showed right coronary artery-pulmonary artery fistula and left coronary circumflexus artery-left atrial multiple fistulae, then the percutaneous coronary intervention spring coil embolization was successfully conducted for right coronary artery-pulmonary artery fistula. After operation, bayasprin enteric-coated tablets 0.1 g/d was taken for anti-platelet aggregation and preventing thrombotic diseases. The observation during operation and postoperative 5-day hospitalization showed no relevant complications. Conclusion Percutaneous coronary intervention is safe and effective for the symptomatic patients with coronary artery-pulmonary artery fistula.
Objective To compare two kinds of myofascial flap encapsulating adi pose-derived stromal cells (ADSCs) in adi pogenic efficacy in vivo, and to provide experimental basis for the efficient transplantation of free adi pose tissue. Methods ADSCs were isolated from the subcutaneous adipose tissue in the neck of 10 New Zealand rabbits (aged 3-4 months old, male and female, weighing 2.0-2.5 kg), and primary culture and subculture of ADSCs were conducted. When the cells at passage 3 covered 70%-80% of the bottom of the culture flask, BrdU (10 μg/mL) was appl ied to label the cells for 48 hours before performing immunofluorescence staining. Oil red O staining observation was conducted to thosecells 2 weeks after being induced towards adi pocyte, al izarin red staining observation was performed 3 weeks after being induced towards osteoblast, and alcian blue staining was conducted 2 weeks after being induced towards chondrocyte. Besides, after being induced towards adipocyte for 2 weeks, 1 × 107 ADSCs/piece at passage 3 labeled by BrdU was seeded into Col I (10 mm × 10 mm × 5 mm/piece) to prepare cell carrier complex. The experiment was divided into two groups: group A in which vascular pedicled dextral latissimus dorsi fascial flap was adopted to encapsulate the complex; group B in which dextral gluteus maximus fascial flap with no specific vessel pedicle was appl ied to encapsulate the complex. Rabbits in each group went through autogenous ADSCs transplant and self control. The implants were dislodged 8 weeks after operation, HE staining and immunohistochemistry staining were performed to testify cambium, the wet weight and micro vessel count of the cambium in each group were tested, immunofluorescence staining was performed to determine the origin of cambium and microvascular endothel ium. Results The nucleus of ADSCs positive for BrdU label ing showed green fluorescence under fluorescence microscope, with the positive label ing ratio of ADSCs above 90%. For ADSCs at passage 3, the formation of red l ipid droplets within cells was observed 2 weeks after being induced towards adipocyte, red calcium nodules were evident 3 weeks after being induced towards osteoblast, and highly congregated cell mass positive for alcian blue staining appeared 2 weeks after being induced towards chondrocyte. Eight weeks after operation, neogenetic blood vessel grew into scaffolds and no obvious fibreencapsulation was observed in group A, while few blood vessel grew into scaffolds in group B. The wet weight of cambium in group A and B was (0.149 5 ± 0.017 3) g and (0.095 3 ± 0.012 7) g, respectively, indicating there was a significant difference between two groups (P lt; 0.01). HE staining showed the formation of neogenetic adipose tissue and the growth of micrangium in the implant, and the degradation and absorption of scaffold. The micro vessel count of group A and B was 31.2 ± 4.5 and 19.3 ± 2.6, respectively, indicating there was a significant difference between two groups (P lt; 0.01). Eight weeks after operation, the immunofluorescence staining of cambium showed that the cell nucleus of regenerated adi pocytes and partial capillary endothel ium in groups A and B presented green fluorescence. Conclusion ADSCs encapsulated by vascular pedicled latissimus dorsi fascial flap and collagen protein scaffold complex has a higher adi pogenic efficacy in vivo than the gluteus maximus fascial flap with no specific vessel pedicle.
Objective To investigate the expression of human leukocyte antigen-G （HLA-G） in hepatocellular carcinoma （HCC） tissues， and to evaluate the prognosis of patients after liver transplantation. Methods The clinical data of 83 patients with HCC who underwent liver transplantation from January 2004 to May 2008 in the Liver Transplantation Center of the Third Affiliated Hospital of Sun Yat-sen University were analyzed retrospectively. The expression of HLA-G in HCC tissues was detected by using immunohistochemical analysis. The Kaplan-Meier method was used to evaluate the cumulative survival rate and tumor-free survival rate. Log-rank test and Cox regression model were used to analyze the single and muti-factor for tumor-free survival rate, respectively. Results Among the 83 patients，there were tumor recurrence in 35 patients （42.2%）. The 1-year，3-year, and 5-year of cumulative survival rate was 97.2%，89.8%, and 43.1%, respectively. The 1-year，3-year， and 5-year of tumor-free survival rate was 93.6%，68.9%, and 38.7%, respectively. The positive rate （68.7%） of HLA-G expression in HCC tissues was significantly higher than that in paracancerous tissues （15.7%）, P＜0.01. A significant association was found between expression of HLA-G and tumor size， vascular invasion, and pathology differentiation （P＜0.05）. Single factor analysis showed that the expression of HLA-G （P＜0.01）， tumor size （P＜0.05）， vascular invasion （P＜0.01）， and pathology differentiation （P＜0.01） effected on tumor-free survival rates of HCC patients after liver transplantation. The tumor-free survival rate in positive expression of HLA-G group was significantly lower than that in negative expression of HLA-G group （P＜0.01）. Cox regression model analysis showed that the expression of HLA-G （P＜0.05）， vascular invasion （P＜0.01）， and pathology differentiation （P＜0.05） were independent risk factors that affected the tumor-free survival rate of HCC patients after liver transplantation. Conclusions There is expression of HLA-G in HCC tissues. The independent risk factors that affecting the tumor-free survival rate of HCC patients after liver transplantation include the expression of HLA-G， vascular invasion, and pathological differentiation. Taking interferential measures for patients with positive expression of HLA-G and strict selection of indication of liver transplantation for HCC can reduce the recurrence rate of tumor.
Objective To construct the expression vector of HLA-G-shRNA and investigate the effect of HLA-GshRNA from NK cell lysis. Methods Four HLA-G shRNA plasmids were constructed and transiently transfected to Bel-7402 cell lines， the levels of mRNA and protein of HLA-G were detected by Real-Time PCR and Western blot. The cytotoxicity of NK-92MI cells against the transfected cells was analyzed by LDH releasing assay. Results The gel electrophoresis and sequencing showed that the inserted sequence was identical to the one which we designed, and no aberrations such as mutation，deletion or insertion occurred. The expressions of HLA-G confirmed by Real Time-PCR and Western blot were significantly down-regulated. Bel-7402 cell lines transfected HLA-G shRNA showed higher lytic activity （P＜0.01）. After KIR2DL4 receptor blocked，lytic activity of NK-92 MI cell were decreased （P＜0.01）. Conclusions HLA-G shRNA plasmids are successfully constructed and HLA-G down-regulated can increase NK cytolysis against Bel-7402 cell. After HLA-G combines with KIR2DL4 receptor at the surface of NK cells， the inhibition effect is transferred.