Genotyping of nucleocapsid protein gene of HCV in HIVHCV co-infected patients in Kunming in 2019 / 中国热带医学

@#Abstract: Objective　To investigate the distribution characteristics of HCV genotypes and subtypes in patients with HIV (human immunodeficiency virus, HIV)/HCV co-infection in Kunming based on the nucleocapsid protein gene sequence of HCV (hepatitis C virus). Methods　Serum was collected from HIV/HCV co-infected patients with household registration in 14 county-level cities, districts and counties under the jurisdiction of Kunming, who admitted to Yunnan Provincial Infectious Disease Hospital from March to August 2019. The viral RNA was extracted from the serum, reverse transcribed to synthesize cDNA, and the HCV nucleocapsid protein gene-specific primers were used for nested PCR amplification. The positive amplification products were sequenced, bioinformatics software such as DNAstar and MEGAX were used for sequence analysis. Results　A total of 64 samples from co-infected patients with clinical diagnosis of suspected HIV/HCV were collected and amplified by HCV nucleocapsid protein gene-specific primers, of which 17 samples were amplified positively. The results of sequence analysis showed that the sequences of 9 cases were located in the same evolutionary branch as the HCV 3b subtype sequence, and the nucleotide homology was 93.3%-95.2%; the sequences of 5 cases were located in the same evolutionary branch as the HCV 1b subtype sequence, and the nucleotide homology was 96.8%-97.6%; the sequence of one case and the subtype sequence of HCV 3a gene were located in the same evolutionary branch, and the nucleotide homology was 95.2%; the sequence of one case and HCV 6n gene subtype sequence were located in the same evolutionary branch, and the nucleotide homology was 97.9%; One case was located in the same evolutionary branch as the HCV 6u gene subtype sequence, and the nucleotide homology was 98.4%. Conclusions　HCV 1b, HCV 3a, HCV 3b, HCV 6n and HCV 6u genotypes or subtypes of HCV are prevalent in Kunming, and HCV 3b is the most prevalent genotype.

Trdmt1 3'-untranslated region functions as a competing endogenous RNA in leukemia HL-60 cell differentiation

Severe blockage in myeloid differentiation is the hallmark of acute myeloid leukemia (AML). Trdmt1 plays an important role in hematopoiesis. However, little is known about the function of Trdmt1 in AML cell differentiation. In the present study, Trdmt1 was up-regulated and miR-181a was down-regulated significantly during human leukemia HL-60 cell differentiation after TAT-CT3 fusion protein treatment. Accordingly, miR-181a overexpression in HL-60 cells inhibited granulocytic maturation. In addition, our "rescue" assay demonstrated that Trdmt1 3′-untranslated region promoted myeloid differentiation of HL-60 cells by sequestering miR-181a and up-regulating C/EBPα (a critical factor for normal myelopoiesis) via its competing endogenous RNA (ceRNA) activity on miR-181a. These findings revealed an unrecognized role of Trdmt1 as a potential ceRNA for therapeutic targets in AML.

Protective Effect of Modified Sinisan on Depression-induced Liver Injury in Rat / 中国实验方剂学杂志

Objective:To study the protective effect of modified Sinisan on liver injury caused by chronic unpredictable mild stress (CUMS) combined with solitary nutrition method based on a depression model. Method:Forty-eight SD male rats were randomly divided into normal group, depression model group, fluoxetine group (1.58 mg·kg-1·d-1), and low, medium and high-dose modified Sinisan (3.67, 7.34, 14.68 g·kg-1·d-1) groups, with 8 rats in each group. After the administration of the model, the levels of alanine aminotransferase（ALT）, aspartate aminotransferase（AST） in serum and the levels of inflammatory factors tumor necrosis factor-α（TNF-α）, interleukin（IL）-1β, and IL-6 in liver tissues were measured by a biochemical kit method. Western blot was used to detect the expressions of cysteine aspastic acid-specific protease-3 (Caspase-3) and cleaved Caspase-3 in liver cells after intervention with modified Sinisan. Result:Compared with the normal group, the biochemical test showed that the levels of ALT and AST in the serum of the model group were significantly increased (P<0.05), and the levels of inflammatory factors TNF-α, IL-1β and IL-6 in liver tissue significantly increased (P<0.05). Compared with the model group, low, medium-dose modified Sinisan groups significantly reduced serum ALT, AST and inflammatory factors TNF-α, IL-1β and IL-6 levels (P<0.05). Western blot showed that compared with the normal group, the protein expression of Caspase-3 in model rat liver cells decreased, while the protein expression of cleaved Caspase-3 increased（P<0.05，P<0.01）. Compared with the model group, low, medium-dose modified Sinisan groups could up-regulate Caspase-3 protein expression, and down-regulated the expression of cleaved Caspase-3 protein（P<0.05，P<0.01）. Conclusion:Chronic stress-induced depression model can cause liver damage. Modified Sinisan can have the hepatoprotective effect by regulating the levels of various physiological, biochemical and inflammatory factor indicators, and inhibiting liver cell apoptosis.

Active substance fractions and effects of Shugan Hewei Tang on hippocampal meurotransmitters in rat model of chronic stress-induced depression / 中国中药杂志

To study the antidepressant effect of Shugan Hewei Tang on chronic stress depression model rats, and select the effective substance fractions. Several male SD rats were randomly divided into 17 groups: blank control group, model group, positive control group(fluoxetine), Shugan Hewei Tang high and low dose groups, 6 high and low dose groups of different substance fractions. After modeling for 3 weeks and administration for 1 week, the effective substance fractions were selected according to the body mass and behavioral performance of SD rats in each group; several neurotransmitters in hippocampus of rats were determined by LC-MS/MS method, including norepinephrine(NE), serotonin(5-HT), 5-indoleacetic acid(5-HIAA), r-aminobutyric acid(GABA), and glutamic acid(Glu). Behavioral results after modeling showed that as compared with the blank group, the body mass growth of the model group was significantly reduced(P<0.01); the sugar water consumption was reduced(P<0.01); the distance between the open field and the number of crossing the central area were also significantly reduced(P<0.01, P<0.01); the resting time was increased significantly(P<0.01); and the forced swimming time was significantly prolonged(P<0.01), indicating that the depression model was effective. After intragastric administration, as compared with the model group, the above detection indicators of volatile oils, total polysaccharides and terpenoid in Fluoxetine, Shugan Hewei Tang groups were all changed(P<0.05 or P<0.01). The levels of NE, 5-HT and GABA in the hippocampus of the model group were significantly lower than those in the blank group(P<0.01), and the levels of 5-HIAA and Glu were significantly increased(P<0.01). As compared with the model group, neurotransmitters of the treatment groups were changed obviously or significantly(P<0.05 or P<0.01). Shugan Hewei Tang showed obvious anti-depressant effects, and the volatile oils, total polysaccharides and terpenoids acted as the main active substances. The mechanism of anti-depression may be related to its regulation of various neurotransmitter levels in the hippocampus.

Effects of Shugan Hewei Decoction and active substance fractions on behavior and neurotransmitter levels in hypothalamus of depression model rats / 中国中药杂志

To investigate the effects of Shugan Hewei Decoction and its active substance fractions on behavior and neurotransmitter levels in hypothalamus of depression model rats,and preliminarily explore its possible mechanism. Male SD rats were randomly divided into blank control group,model group,fluoxetine( positive control) group,Shugan Hewei Decoction high and low dose groups,high and low dose groups of three different substance fractions. After 3 weeks' CUMS and social isolation to induce models,intragastrical administration lasted for 7 d. Behavioral experiments( sucrose consumption test,open-field test,and forced swimming test) were then performed to evaluate the depression status of rats. Several neurotransmitters in hypothalamus of rats were determined by LC-MS/MS method,including dapamine( DA),norepinephrine( NE),serotonin( 5-HT),5-indoleacetic acid( 5-HIAA),γ-aminobutyric acid( GABA),and glutamic acid( Glu). As compared with the blank control group,the sucrose consumption was reduced( P<0. 01); the total distance and the number of crossing the central area were also significantly reduced( P< 0. 01,P< 0. 01),while the resting time increased significantly( P<0. 01); the forced swimming time was significantly prolonged( P<0. 01); DA,5-HT,NE,5-HIAA and GABA levels in hypothalamus were significantly reduced( P < 0. 01),while Glue level was significantly increased( P < 0. 01) in model group. As compared with the model group,all the above indexes had changes in fluoxetine group,Shugan Hewei Decoction whole recipe groups,volatile oils group,polysaccharides group,and terpenoids group( P<0. 01 or P<0. 05). Shugan Hewei Decoction whole recipe and its active substance fractions can improve the behavior of depression model rats and may exert anti-depression effects by regulating the content of neurotransmitters in the hypothalamus.

Endoscopic treatment for ureterovesical junction stenosis in patients with kidney transplantation: 13 cases report / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the safety and efficacy of endoscopic treatment for ureterovesical junction (UVJ) stenosis in patients with kidney transplantation.@*METHODS@#A retrospective study was conducted among the patients with kidney transplantation diagnosed as UVJ stenosis from 2012 March to 2018 July in Urology and Lithotripsy Center, Peking University People's Hospital. Only the patients who received endoscopic treatment were included, with staged or same-session nephrostomy followed by a retrograde ureteroscopy to evaluate the ureteral stenosis. Incisions with laser, mono- or bipolar energy, or balloon dilation were used to manage the stenosis depending on different situations. Demographic characteristics and clinical data were gathered and analyzed, including age, gender, preoperative serum creatinine, hemoglobin, operation time, success rate, postoperative serum creatinine, hemoglobin, postoperative complications rate, and long-term stenosis recurrence rate.@*RESULTS@#In this study, 13 patients were included (9 males and 4 females). All the UVJ stenoses were diagnosed with preoperative ultrasound, CT scan, MRI, or urethrography. The mean age was 45 years (range 34-57 years). The mean preoperative serum creatinine was 243 μmol/L. Four patients developed UVJ stenosis 1 month after kidney transplantation, while the rest developed long-term stenosis. Fifteen operations were performed in all, of which 14 cases were successful while one failed. The first 8 cases received first-stage nephrostomy and second-stage endoscopic management of the stenosis, while the last 7 cases received the same session surgery. The mean operation time was 95.4 min vs. 68.9 min, and the immediate success rate was 87.5% vs. 100.0% in the first 8 cases and last 7 cases, respectively. The mean decrease of postoperative hemoglobin was 0.6 g/L and mean postoperative serum creatinine was 105 μmol/L. No postoperative fever, severe hematuria, and urine leak were observed. The mean postoperative hospital stay was 2.8 days. Three patients were able to remove ureteral stents and no recurrence was found with a follow-up time of 9, 17, and 82 months. The long-term stenosis recurrence rate was 76.9% (10/13).@*CONCLUSION@#Endoscopic approach for the treatment of UVJ stenosis in patients with kidney transplantation was safe and efficient in our study cohort. However, long term stenosis recurrence rate was high and needed to be paid attention to.

Treatment of crossing vessels in laparoscopic pyeloplasty / 北京大学学报(医学版)

OBJECTIVE@#To investigate the management of crossing vessels compression in laparoscopic pyeloplasty.@*METHODS@#From January 2016 to June 2018, a total of 21 patients who were admitted to Peking University People's Hospital with ureteropelvic junction obstruction (UPJO) associated with crossing vascular compression were reviewed. There were 15 males and 6 females who formed this group, with a mean age of (33.9±15.0) years. There were 4 cases of mild hydronephrosis, 12 cases of moderate hydronephrosis and 5 cases of severe hydronephrosis before operation. All the patients underwent laparoscopic pyeloplasty in our hospital, including 13 on the left and 8 on the right. Laparoscopic pyeloplasty (Anderson-Hynes) were performed in all the patients. Hem-o-lok suspension (14 cases in the suspension group) or translocation of the crossing vessels (7 cases in the translocation group) were used for the intraoperative management of the crossing vessels. Double J tubes were removed 8 weeks postoperatively. The patient demographic data were collected (including operation time, treatment time of crossing vessels, intraoperative blood loss, time of drainage tube removal after operation, and average length of hospital stay), postoperative outcomes were evaluated and the patients were followed up regularly.@*RESULTS@#In all the patients, the crossing vessels were successfully reserved, and none of them were ligated intra-operatively. Mean operative times were (202.2±57.0) min. The duration of intraoperative treatment of crossing vessels was (10.5±3.2) min, (6.1±2.0) min in the suspension group, and (13.7±5.2) min in the translocation group, respectively. Intraoperative blood loss was (47.8±25.6) mL, postoperative drainage time was (4.8±2.6) d, and length of hospital stay was (11.5±3.3) d. Postoperative slight urinary leakage occurred in 1 case. Preoperative pyelectasis of the affected side of all the patients was (3.4±1.7) cm, compared with postoperative pyelectasis of (1.9±1.3) cm. The difference was statistically significant (P<0.05). Postoperative follow-up of all the patients was carried out until December 2018. There was no significant difference in kidney size in all the patients before or after the operations, and hydronephrosis was alleviated compared with that before surgery.@*CONCLUSION@#For UPJO patients with crossing vascular compression, according to the location of the crossing vessels, Hem-o-lok suspension or vessel transposition can be adopted to relieve the crossing vascular compression and improve the success rate of the surgery.

Review of upper urinary modified minimal invasive surgical technology / 北京大学学报(医学版)

Upper urinary surgery is an important area of urology surgery. Open surgery used to be the gold standard of upper urinary surgery. With the development of medical techniques, minimal invasive surgeries including laparoscopic and robot assisted-laparoscopic surgery have gradually replaced the open surgery. Because of the complexity and diversity of upper urinary diseases, surgeries sometimes are difficult, and minimal invasive surgeries require higher surgical abilities of urologists than open surgeries. In recent years, depending on our surgical experience and international reports, our team from three Chinese medical centers summarizes techniques of upper urinary minimal invasive surgeries. For malignant diseases, such as renal and ureteral carcinomas, it's important to totally remove the tumor first, and then to avoid the surgical injuries. We summarize surgical experience of retroperitoneal laparoscopic partial nephrectomy for moderately complex renal hilar tumors. Our team modified minimal invasive techniques for some complex tumors, including ring suture technique for renal hilar tumors, internal suspension technique for renal ventral tumors, and combination retroperitoneal laparoscopic surgery with mini-flank incision for complex renal tumors. While for begin diseases, urologists should focus on the resections and surgical injuries at the same time. We have reported the novel technique of laparoscopic aspiration for central renal angiomyolipoma, making the surgery simple and available. For reconstruction surgeries, operations should be based on several principals. We generalize it as "4TB principals", which include "tension-free", "water-tight", "thin suture", "no touch of the key area" and "protecting the blood supply". Depending on the localization, length, and etiology of the strictures, different techniques are required. Our team summarize the pyeloplasty, ureteral reimplantation and ileal ureter replacement based on our surgical experience. For infant upper urinary surgeries, our team has made invasive surgeries that can be used in complex diseases, such as duplex kidney. Based on years of surgical techniques, our modified surgeries achieve a better subjective cosmetic result than the traditional surgeries. In the future, the standardized, practical, simple and individual minimal invasive surgical technique will become the main direction in the future researches.

Preparation and identification of protein chip for detecting components of peach allergen / 基础医学与临床

Objective To prepare a protein chip for the detection of serum antibodies against several peach allergen components, and to provide a rapid and reliable method for clinical diagnosis of peach allergic disease .Methods The coding sequences of six key peach components Pru p 1, Pru p 2, Pru p 3, Pru p 4, Pru p 7 and Pru p G were inserted into pGAPZαA yeast expression vector.Then the vectors were digested by endonuclease Avr Ⅱ and electroporated into yeast SMD1168 for protein expression.After peach proteins were purified, the protein chip was prepared and used to detect the antibodies against peach allergen components in serum samples from 41 suspected patients.The sensitivity and specificity of the protein chip were verified by comparison with ImmunoCAP method .Res ults The protein chip was prepared with these proteins for detecting IgE antibodies in serum samples against these four peach allergen components.The results showed that the sensitivity of the protein chip to Pru p 1, Pru p 3 and Pru p 4 was 40%, 100%and 100%, and the specificity was 100%, 50%and 100%respectively.The total sensitivity and specificity was 86%and 96%respectively.Six serum samples from peach allergy patients were identified Pru p7 antibody positive by the protein chip.Co nclusions The sensitivity and specificity of the protein chip is comparable to ImmunoCAP.It needs less serum and so to be a potential method for the clinical diagnosis of peach allergenic disease after optimization.

Study on the effect of single dose intraosseous injection of simvastatin on tumor vascular normalization in murine breast cancer / 中国比较医学杂志

Objective To investigate the effect of single dose intraosseous injection of simvastatin on tumor vascular structure and function in murine breast cancer. Methods BALB/c mice and 4T1 murine breast cancer cells were used to establish a subcutaneous xenograft model. The mouse model of orthotopic breast cancer received intraosseous injection of a single dose of simvastatin (50 μg) or vehicle only. Frozen tumor tissue sections were prepared for co-immunostained with CD31 andα-SMA. Evans blue dye was injected into the tail vein to observe the vascular permeability. The expression level of HIF-1αwas detected by immunohistochemistry. Results Immunofluorescence dual staining showed that intraosseous injection of simvastatin increased the number of perivascular pericytes in the tumor vessel(P < 0. 05), Evans blue dye content showed that in vivo vessel permeability in the tumor tissue was significantly decreased(P < 0. 05), and the immunohistochemistry results showed that local hypoxic area was significantly improved. Conclusions Single dose intraosseous injection of simvastatin can promote the normalization of tumor vasculature by improving the coverage of pericytes.

Effect of single intraosseous injection of simvastatin on neoangiogenesis in myocardial infarction in rats / 中国实验动物学报

Objective To explore the effect of single local intraosseous injection of small dose simvastatin on the angiogenesis and cardiac function in rats after myocardial infarction. Methods Adult male Wistar rats were divided into sham operation group, myocardial infarction model group and intraosseous injection of simvastatin 0. 5 mg group ( all n=12 per group) . The left anterior descending branch of coronary artery was ligated to establish a rat model of myocardial infarc-tion. The left ventricular function was evaluated by small animal echocardiography at 4 weeks postoperatively. The rest of the rats were sacrificed, the myocardial infarct size was evaluated by TTC staining, and the myocardial neovascularization was detected by immunofluorescence staining. Results We successfully established the rat model of myocardial infarction. The echocardiography showed that the left ventricular systolic function was decreased significantly at 4 weeks after myocardi-al infarction. Intraosseous injection of simvastatin (0. 5 mg) did not improve the left ventricular function after myocardial infarction in the rats. TTC staining showed that intraosseous injection of simvastatin did not reduce myocardial infarct size. Immunofluorescence staining showed that the myocardial capillary density of simvastatin group was slightly higher than that of myocardial infarction model group, but showing no significant difference between them. Conclusions Intraosseous in-jection of simvastatin 0. 5 mg 24 hours after myocardial infarction cannot significantly promote myocardial angiogenesis, which is believed to be beneficial to the revascularization after ischemia, and thus failed to improve the cardiac function.

Study on the effect of single dose intraosseous injection of simvastatin on tumor vascular normalization in murine breast cancer / 中国比较医学杂志

Objective To investigate the effect of single dose intraosseous injection of simvastatin on tumor vascular structure and function in murine breast cancer. Methods BALB/c mice and 4T1 murine breast cancer cells were used to establish a subcutaneous xenograft model. The mouse model of orthotopic breast cancer received intraosseous injection of a single dose of simvastatin (50 μg) or vehicle only. Frozen tumor tissue sections were prepared for co-immunostained with CD31 andα-SMA. Evans blue dye was injected into the tail vein to observe the vascular permeability. The expression level of HIF-1αwas detected by immunohistochemistry. Results Immunofluorescence dual staining showed that intraosseous injection of simvastatin increased the number of perivascular pericytes in the tumor vessel(P < 0. 05), Evans blue dye content showed that in vivo vessel permeability in the tumor tissue was significantly decreased(P < 0. 05), and the immunohistochemistry results showed that local hypoxic area was significantly improved. Conclusions Single dose intraosseous injection of simvastatin can promote the normalization of tumor vasculature by improving the coverage of pericytes.

Effect of single intraosseous injection of simvastatin on neoangiogenesis in myocardial infarction in rats / 中国实验动物学报

Objective To explore the effect of single local intraosseous injection of small dose simvastatin on the angiogenesis and cardiac function in rats after myocardial infarction. Methods Adult male Wistar rats were divided into sham operation group, myocardial infarction model group and intraosseous injection of simvastatin 0. 5 mg group ( all n=12 per group) . The left anterior descending branch of coronary artery was ligated to establish a rat model of myocardial infarc-tion. The left ventricular function was evaluated by small animal echocardiography at 4 weeks postoperatively. The rest of the rats were sacrificed, the myocardial infarct size was evaluated by TTC staining, and the myocardial neovascularization was detected by immunofluorescence staining. Results We successfully established the rat model of myocardial infarction. The echocardiography showed that the left ventricular systolic function was decreased significantly at 4 weeks after myocardi-al infarction. Intraosseous injection of simvastatin (0. 5 mg) did not improve the left ventricular function after myocardial infarction in the rats. TTC staining showed that intraosseous injection of simvastatin did not reduce myocardial infarct size. Immunofluorescence staining showed that the myocardial capillary density of simvastatin group was slightly higher than that of myocardial infarction model group, but showing no significant difference between them. Conclusions Intraosseous in-jection of simvastatin 0. 5 mg 24 hours after myocardial infarction cannot significantly promote myocardial angiogenesis, which is believed to be beneficial to the revascularization after ischemia, and thus failed to improve the cardiac function.

Look into Hepatic Progenitor Cell Associated Trait: Histological Heterogeneity of Hepatitis B-Related Combined Hepatocellular-Cholangiocarcinoma / 华中科技大学学报（医学）（英德文版）

Combined hepatocellular-cholangiocarcinoma (CHC) is a mixed tumor containing elements of both hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC).Its remarkable histological heterogeneity has been linked to putative hepatic progenitor cell (HPC) origin.However,detailed histological or phenotypic description is rarely documented.In the present study,we reassessed 68 cases previously diagnosed as hepatitis B-related CHCs by immunohistochemistry and double-fluorescence immunostaining,focusing on HPC associated phenotypic observation of intermediate area of the tumor.It was found that tumor cells showed remarkable heterogeneity in intermediate area.Tumor cells with intermediate morphology between hepatocytes and cholangiocytes were oval-shaped and small with scant cytoplasm and hyperchromatic nuclei,arranging in solid nests mostly.By Keratin 7 (K7) staining,it appeared that the nests of tumor cells represented a maturation process from the undifferentiated small cells to mature hepatocytes through the "transitional" cells.Then,these small cells were further confirmed with intermediate phenotype as HPC by exploring immature hepatocellular marker and HPC/biliary markers co-localization.In conclusion,the HPC associated trait in CHC can be interpreted by HPC origin or gain of"stemness" by dedifferentiation.It is still too soon to give a final word that it is innate or acquired signature of HPC associated trait in CHC.

Analysis and clinical implication of upper urinary tract stone's bacterial spectrum / 中华外科杂志

<p><b>OBJECTIVES</b>To analyze the pathogenic bacterial's distribution and the drug resistance in the upper urinary tract stones, and to provide the information for choosing suitable antibiotics.</p><p><b>METHODS</b>Stone samples were taken for culture and for drug sensitivity test in 146 patients undergoing percutaneous nephrolithotomy between April 2007 and October 2008, and the results were analyzed.</p><p><b>RESULTS</b>Pathogens presented in 72 (49.3%) patients. There were 70 (86.4%) Gram-negative bacteria strains. Escherichia coli, Pseudomonas aeruginosa and Enterobacter cloacae were the predominant bacteria, accounted for 30.9%(25 strains), 23.5% (19 strains) and 12.3% (10 strains), respectively. There were 10 (12.3%) Gram-positive bacteria strains, the predominant bacteria was Staphylococcus epidermidis (6 strains), accounting for 7.4%. And there was 1 fungi strain (1.2%). Resistance to ampicillin/sulbactam (88.7%), ceftriaxone (81.3%) and ciprofloxacin (67.5%) was most commonly found in pathogen, and the rate of resistance to amikacin, imipenem and piperacillin/tazobactam were 8.6%, 10.0%, 10.0%, respectively. Erythromycylamine, teicoplanin, SMZ-TMP, nitrofurantoin were sensitive to Gram-positive bacteria.</p><p><b>CONCLUSIONS</b>Bacterial's distribution of upper urinary tract stones are multiple, and the majority pathogen is Gram-negative bacteria. A big variant resistance is found among different bacterium. The suitable antibiotics should be chosen according to the different bacterium in the patients who underwent percutaneous nephrolithotomy.</p>

Construction and screening of suppression subtractive hybridization library of renal cell carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To construct and screen the suppression subtractive hybridization (SSH) library of human renal cell carcinoma (RCC).</p><p><b>METHODS</b>Poly A(+) RNA was isolated from RCC lines 786-O (tester) and renal cell (RC) lines HK-2 (driver), respectively. SSH procedure was performed according to the protocol of the PCR-Select cDNA Subtraction Kit (Clontech), and PCR products were cloned into pT-Adv vector and transformed E. coli TOP10F'. All positive clones picked out were digested and some of which were sequenced.</p><p><b>RESULTS</b>The SSH library contained 362 clones with SSH cDNA fragments distributed mainly from 0.3 to 0.9 kb. Among 50 clones sequenced randomly, 2 represented unknown genes and the other 48 derived from 36 known genes.</p><p><b>CONCLUSION</b>The quality of the SSH library of human RCC is reliable and its construction is the basis for further screening differentially expressed genes of RCC.</p>

Clinical application of endovascniar stent-graft in the treatment of portal stenosis of cancerous thrombus / 中华放射学杂志

Objective To explore the approach and early effects of endovascular stent-graft deployment in the treatment of portal stenosis of cancerous thrombus.Methods Six cases with portal vein stenosis of cancerous thrombus,which caused by primary hepatic carcinoma(5 cases)and eholangiocarcinoma(1 case)and the severity of stenosis showed on contrast enhanced CT were more than 75% or occluded,were performed percutaneous transhepatie or transsplenic portography.FLUENCY~(TM) endovascular stent-graft(10 mm diameter)was placed at the position of stenosis after gastroesophageal varices embolization.Portal pressure was measured pre-and post-deployment.Results Stents were successfully placed in all patients.The average portal pressure decreased from 50.7 cm H_2O(1 cm H_2O = 0.098 kPa)to 41.3 cm H_2O after endovascular stent-graft deployment.The restenosis were found in 2 cases after one month.Haematemesis and refractory aseites appeared in one case respectively,the other 4 cases showed no significant symptoms above caused by portal hypertension.Conclusion It is safe and feasible for endovaseular stent-graft deployment in the treatment of portal stenosis of cancerous thrombus.Selecting the suitable indications,the symptoms of portal hypertension can be controlled effectively.

Effects of dominant-negative truncation mutant ?NTCF4 on biological characteristics of renal cancer cell line GRC-I by down-regulation Wnt signaling pathway target genes / 中华泌尿外科杂志

Objective To investigate the effects of dominant-negative truncation mutant?NTCF4, lacking the N-terminal form of TCF4 gene,on biological characteristics of renal cancer cell line GRC-I and explore the molecular mechanisms.Methods GRC-I cell was transfected with pCDNA3-?NTCF4 eukary- otie expression plasmid,pCDNA3 empty vector to construct the stable cell line GRC-I/?NTCF4 and GRC-I/ Mock respectively.The morphological changes of stable cells were observed and the cells growth curve was detected through light microscope.The cellular proliferation activities were determined using the MTT assay. The protein expression of Wnt pathway downstream target gene C-Myc and Cox-2 was evaluated by immuno- cytoehemieal method and Western Blot analysis.Results After the dominant-negative?NTCF4 gene was permanently expressed,the GRC-I/?NTCF4 stable cells morphologically showed that appearance changed from circular to long-spindle shape,growth rate decreased with less karyosehisis found,malignant pheno- types reversed to normal renal tubular cells.MTT assay revealed that the proliferation activities of GRC-1/?NTCF4 cells were inhibited by 11.2%-35.5% compared with GRC-I cells (P＜0.05),while the GRC- I/Mock cells have no difference with the control cells.Immunocytochemical analysis and Western Blot showed that the C-Myc and Cox-2 protein expression level of GRC-I/?ANTCF4 cells were significantly sup- pressed in comparison with that of GRC-I/Mock and GRC-I cells.Conclusions The dominant-negative truncation mutant?NTCF4 could partially inhibit the growth of renal cancer cells and down-regulate the pro- tein expression of Wnt pathway target gene C-Myc and Cox-2.These findings provide a experimental founda- tion for applying cell signal therapy to renal cell cancer by blocking the Wnt signaling pathway.