Objective To explore the effect of wild type or mutant parkin gene expression on the growth of human hepatocellular carcinoma cell line Huh-7. Methods The parkin (wild type or mutant) expression vector and empty vector were transferred into Huh-7 cell lines with LipofectAMINE 2000 reagents. The positive clones that expressed parkin gene stably were chosen by G418 and checked by reverse transcription-polymerase chain reaction (RT-PCR) to check the DNA sequences. The cytobiological behaviors of those positive clones were analyzed by cell proliferation assay and tumorigenesis in nude mice. Results Huh-7 cell lines that expressed wild type or mutant parkin gene stably were successfully established. The growth of wild type parkin-expressed cells was obviously inhibited compared with the control cells transfected with empty vectors(t= 3. 875, P= 0. 031).The volume of tumor formed by wild type parkin-expressing cells in nude mice was also significantly reduced (t=8. 228,P=-0. 003). Mutant parkin gene expression had a slight effect on the growth of Huh-7 cells in vitro and in vivo (P＞0.05). Conclusion The re-expression of wild type parkin gene can favor the malignant phenotype revision of Huh-7 cells. Therefore, it might be a good candidate for tumor suppressor gene associated with HCC.

OBJECTIVE: To investigate the position change of the fibular bone after maxillary reconstruction by free fibular flap and to analyze the factors affecting the position change. METHODS: Patients who underwent maxillary reconstruction by free fibular flap in the Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology from November 2012 to November 2016 were enrolled in this study. CT scans 1 week and 1 year postoperatively were collected and stored in DICOM format. The ProPlan CMF software was used to reconstruct the CT scans and separate the maxilla and each segment of the fibular flap. The Geomagic Control software was used to measure the long axis direction vector of each fibular segment. And the position change direction was recorded. The patients were divided into groups according to the use of the fibula or titanium plate to reconstruct the zygomaticmaxillary buttress. RESULTS: A total of 32 patients were enrolled. Among them, 21 were in the titanium plate group and 11 in the fibula bone group. The angle between the long axis of the fibular segment and the X axis in the X-Y plane was 95.65°±53.49° and 95.53°±52.77°, 1 week and 1 year postoperatively, and there was no statistical difference (P>0.05). The angle between the long axis of the fibular segment and the X axis in the X-Z plane was 96.88°±69.76° and 95.33°±67.42°, respectively, with statistical difference (P=0.0497). The angular changes of the long axis of the fibular segment in the titanium plate group and the fibular bone group were 3.23°±3.93° and 1.94°±1.78°, respectively, and the angular changes in the X-Z plane were 6.02°±9.89° and 3.27°±2.31°, respectively. There was no significant difference between the groups (P>0.05). The long axis changes of the fibular segment in the X-Y plane for reconstruction of the anterior alveolar, posterior alveolar, and buttress were 3.13°±3.78°, 2.56°±3.17°, and 5.51°±4.39°, respectively. There was a statistical difference (P = 0.023) between the posterior and buttress. In the X-Z plane, theses were 4.94°±4.75°, 5.26°±10.25°, 6.69°±6.52°, respectively. There was no statistical difference among the three groups (P>0.05). The main positional deviation directions of the titanium plate group and the fibular bone group were interior and superior sides, and there was no statistical difference between the two groups (P>0.05). CONCLUSION One year postoperatively, the position of the free fibular flap was changed compared with 1 week postoperatively. The position of the free fibular flap was mainly changed to the interior and superior sides.
Bone Transplantation ; Fibula/diagnostic imaging* ; Free Tissue Flaps ; Humans ; Mandibular Reconstruction ; Maxilla/surgery*

Glomerular hypertrophy is the main pathological characteristic in the early stage of diabetic nephropathy (DN), and its regulatory mechanism is closely related to mammalian target of rapamycin (mTOR) signaling pathway activity. mTOR includes mTOR complex 1 (mTORC1) and mTOR complex 2(mTORC2), in which, the upstream pathway of mTORC1 is phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase(Akt)/adenosine monophosphate activated protein kinase(AMPK), and the representative signaling molecules in the downstream pathway of mTORC1 are 4E-binding proteins(4EBP) and phosphoprotein 70 S6Kinase(p70S6K). Some Chinese herbal extracts could improve cell proliferation via intervening the expressions of the key molecules in the upstream or downstream of PIK/Akt/mTOR signaling pathway in vivo. As for glomerular mesangial cells(MC) and podocyte, mTOR plays an important role in regulating glomerular inherent cells, including adjusting cell cycle, energy metabolism and matrix protein synthesis. Rapamycin, the inhibitor of mTOR, could suppress glomerular inherent cell hypertrophy, cell proliferation, glomerular basement membrane (GBM) thickening and mesangial matrix deposition in model rats with DN. Some Chinese herbal extracts could alleviate glomerular lesions by intervening mTOR signaling pathway activity in renal tissue of DN animal models or in renal inherent cells in vivo and in vitro.
Animals ; Diabetic Nephropathies ; drug therapy ; enzymology ; genetics ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertrophy ; drug therapy ; enzymology ; genetics ; pathology ; Kidney Glomerulus ; drug effects ; metabolism ; pathology ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism

Objective　To evaluate the role of transfected angiotensinⅡ(Ang Ⅱ) receptor AT1 anti-sense nucleotide (AT1A) in the expression of subtypes of AngⅡ receptor mRNA, synthesis of protein and nucleic acid in cardiomyocytes. Methods　AT1 cDNA sequence (476 bp) was cloned with RT-PCR and reversely inserted into PcDNA3.1 (5.4 kb) to construct an intact plasmid containing AT1A (PAT1A). The plasmid was then transfected into the cultured cardiomyocytes and identified with RT-PCR and Western blot. The synthesis of protein and nucleic acid identified by 3H-Leu and 3H-TdR incorporation, and expressions of AT1 and AT2 mRNA by RT-PCR, were compared between transfected and nontransfected cardiomyocytes after being stimulated with 10-7 mol/L AngⅡ for 24 h. Results　The plasmid PAT1A were successfully constructed. The AT1 mRNA and its protein were expressed significantly less in the transfected cardiomyocytes than in the control (P<0.01). In the transfected cardiomyocytes, AT1 mRNA expression was markedly decreased, but that of AT2 mRNA obviously increased (P<0.01) when compared with the nontransfected cardiomyocytes after stimulation for 24 h with AngⅡ 10-7 mol/L; no significant difference was found in 3H-Leu and 3H-TdR incorporation between them. Conclusion　After the cardiomyocytes was tranfected with AT1A, the expression of AT1 mRNA was markedly suppressed，while AT2 mRNA up-regulated at the same time. Our results indicate that AT1A blocking can not effectively interrupt the Ang Ⅱ-induced synthesis of the protein and nucleic acid in cardiomyocytes.

Objective To investigate the effects of Xiaoyu Sanjie Yi'ai Enema on the expression level of phosphatase and tensin homolog deleted on chromosone 10(PTEN)gene of nude mouse model of prostate cancer, and to explore its anticancer mechanism. Methods Nude mouse model of prostate cancer was established by neck subcutaneous injection of suspension of human prostate cancer cells PC-3. Thirty tumor-bearing nude mice were randomly divided into control group and treatment group , 15 mice in each group. Ten days after modeling, the treatment group was treated with enema of Xiaoyu Sanjie Yi'ai Enema, and the control group was treated with enema of the same volume of normal saline,once every other day,for 30 continuous days. At the end of medication, the tumors were extracted completely, and then the tumor mass of the two groups was detected before and after treatment to calculate the tumor-inhibition rate. The mRNA and protein expression levels of PTEN gene were detected by the method of reverse transcription real-time quantitative polymerase chain reaction(RT-qPCR)and western blotting method,respectively. Results Tumor mass in the treatment group was reduced as compared with that in the control group (P < 0.05). The tumor-inhibition rate of the treatment group was 18.71%. RT-qPCR results showed that the relative expression level of the PTEN mRNA in the treatment group was higher than that in the control group, the difference being statistically significant (P < 0.001). Western blotting result showed that the relative protein expression level of PTEN in treatment group was (0.55 ± 0.13)and that in the control group was(0.32 ± 0.09)separately,the difference being statistically significant(P<0.001). Conclusion Xiaoyu Sanjie Yi'ai Enema has therapeutic effects on prostate carcinoma,and the molecule mechanism may be related with the up-regulation of PTEN gene mRNA expression and protein translation,thus to inhibit PC-3 cell proliferation and metastasis.

Objective To observe the effect of splenectomy on mortality and brain water content of rats with brain injury so as to explore novel way for better clinical management of patients with severe traumatic brain injury. Methods Adult male SD rats were randomly divided into three groups, ie, sham operation on brain and spleen (Group A, n = 23), experimental brain trauma & sham operation on spleen (Group B, n =48) and experimental brain injury & splenectomy (Group C, n = 47). Modified Feeney' s method was used to create the animal model of experimental brain trauma, Longa' s scale was applied to evaluate the neurologic defect. Mortality within seven days following brain injury was calculat-ed. In the meantime, the brain water content was detected at days 1 (n = 8), 2 (n = 8), 3 (n = 8) and 7 (n = 7) after brain injury in each group, Results No statistical difference of Longs' s scale was found between Group B and Group C (P > 0.05). The mortalities within seven days after brain injury were 0%, 35.42 and 14.89% in Groups A, B and C respectively, with statistical difference between groups (P<0.05). The brain water content of Groups B and C at days 1, 2, 3 and 7 were (81.98±0.35)% & (81.78±0.41)%, (82.58±0.63)% & (81.81±0.48)% (P<0.05),(82.54±0.54)% & (81.52±0.84)% (P<0.05) and (81.50±0.41)% & (81.21±0.36)% (P>0.05) respectively. Conclusion Splenectomy can effectively reduce brain water content and significantly decrease mortality in rata with brain injury.

OBJECTIVESTo study the etiology of persistent hematospermia and to evaluate the efficacy of transrectal ultrasonography (TRUS)-guided transperineal needle aspiration and irrigation for diagnosis and treatment of persistent hematospermia.

METHODSTwelve patients were included in the study, with a mean age of (36.4 +/- 10.8) years old, and a mean duration of the disease of (13.9 +/- 6.4) months. After the expressed prostatic secretion (EPS) by prostatic massage was cultured, patients with recurrent hematospermia received TRUS-guided transperineal needle aspiration for seminal vesicle fluid (SVF), which was sent for bacteriological and cytological examination. If the EPS culture were positive, certain antibiotics according to the drug sensitivity assay were injected into the abnormal seminal vesicle(s) via TRUS-guided transperineal needle puncture. The treatment would be repeated one month later if the patients still had hematospermia. The patients were followed up every three months.

RESULTSAbnormal images were found in left seminal vesicle (SV) in 4 cases, right in 3 cases, bilateral in 2 cases, and no abnormal findings in 3 cases. The abnormal findings included: 7 cases of SV and/or ejaculatory duct dilation, 3 cases of thickening SV wall, 3 cases of calcification or calculi of SV, and 1 case of Müllerian duct cyst. SVF cultures were positive in 7 cases: methicillin-resistant Staphylococcus aureus (MRSA) 4 cases, methicillin-resistant coagulase-negative Staphylococcus (MRCNS), E. Coli, Proteus mirabilis 1 case, respectively. In five of these 7 cases, bacteriological cultures of SVF and EPS showed the same results. All patients were treated by TRUS-guided transperineal injection of certain antibiotics into SV. Seven cases were injected once, 5 cases twice. The mean follow-up period of 10 patients was (16.7 +/- 5.9) months. Hematospermia disappeared in 6 cases.

CONCLUSIONSSV infection of bacteria, especially infection of the drug resistant strains was one of the main causes of persistent hematospermia. The difficulties in treatment of persistent hematospermia were due to infection of drug resistant bacteria, calcification or calculi of SV, obstruction of ejaculatory duct. TRUS-guided transperineal aspiration of SVF was helpful to the etiologic diagnosis of persistent hematospermia.

Adult ; Drug Resistance, Bacterial ; Genital Diseases, Male ; etiology ; microbiology ; Humans ; Male ; Middle Aged ; Pilot Projects ; Seminal Vesicles ; microbiology ; Ultrasound, High-Intensity Focused, Transrectal

BACKGROUNDThe abnormalities of coronary arteries, though rare and sometimes benign, may first present clinically as myocardial infarction or sudden death. Multi-detector computed tomography (MDCT) is a non-invasive test that is highly suitable for detecting these anomalies. The study aimed to review the 64-MDCT appearance of the coronary artery anomalies in 66 patients and to discuss the clinical importance of these anomalies.

METHODSIn 6014 consecutive patients examined over 12 months by 64-MDCT for the study of coronary artery disease, 66 were diagnosed for coronary artery anomalies. All patients were symptomatic for one or more of the following diseases: chest pain, dyspnoea, palpitations, arrhythmia and myocardial infarction. Nine patients had undergone a coronary angiography. All the CT images were evaluated by two radiologists and one cardiologist. The right coronary artery (RCA) and the conus branch arising separately, myocardial bridging and duplication of arteries were not analysed in our study.

RESULTSThe incidence of coronary artery anomalies found in our study group was 1.097%. In the selected patients, seven different types of coronary anomalies were found by 64-MDCT examination. The high takeoff, origin of the coronary artery from the opposite or noncoronary sinus with an anomalous course, and coronary artery fistula were the three common forms of anomalies (n = 16, 18 and 16, respectively). Compared with the results of the coronary angiography, the number of the drainage sites of two coronary artery fistula was less in MDCT images (3 small sites in total). In all cases, coronary artery computed tomography angiography (CTA) technique was able to recognize the origin of the coronary artery, its three-dimensional course and its spatial relationship with the adjacent structures. Conventional coronary angiography in two cases, however, was unable to provide sufficient information for correct and complete diagnosis.

CONCLUSIONSIn conclusion, the study showed that 64-MDCT, especially the volume rendering technique (VRT), may be useful for the assessment of complex variations, even if the conventional angiography may not be sufficient. It may be considered as the first-choice imaging modality when an anomalous coronary artery is suspected.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Coronary Vessel Anomalies ; diagnostic imaging ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Young Adult

Objective To study antimicrobial resistance and genotyping of methicillin-resistant Staphylococcus au-reus(MRSA). Methods A total of 967 no-repetitive strains of Staphylococcus aureus(S.aureus)isolated from a hospital between January 2014 and November 2015 were collected,antimicrobial susceptibility testing,mecA gene,and Panton-Valentine leukocidin gene(PVL gene)were detected;staphylococcal cassette chromosome mec(SCCmec)typing,multilocus sequence typing(MLST),S.aureus protein A(spa)gene typing,and S.aureus ac-cessory gene regulator(agr)typing were performed with multiplex polymerase chain reaction. Results Of 967 strains of S.aureus,210(21.72%)were MRSA;detection rate of MRSA from sputum specimen was higher than that of skin and soft tissue specimen(68.09% vs 1 1.83% ,P<0.05);vancomycin- and linezolid-resistant S.aureus strains were not found,susceptibility rates of MRSA to gentamicin,tetracycline,erythromycin,clindamycin,levo-floxacin,ciprofloxacin,moxifloxacin,nitrofurantoin,and rifampicin were all lower than those of methicillin-sensi-tive Staphylococcus aureus(MSSA),differences were all statistically significant(all P<0.05);antimicrobial sus-ceptibility rate of MRSA to compound sulfamethoxazole was higher than MSSA,difference was significant(P<0.05). Susceptibility rates of MRSA isolated from skin and soft tissue to gentamicin,levofloxacin,ciprofloxacin,moxifloxacin,and rifampicin were 86.90% -95.24%,while MRSA isolated from sputum were only 1.56% -15.63%.Of 967 strains of S.aureus,210 harbored mecA gene,10 harbored PVL gene,8(3.81%)of 210 MRSA strains weren't typed. The main types of MLST,SCCmec,spa,and agr were ST 239(n= 177 strains),type Ⅲ(n= 177 strains),t 030(n= 177 strains),and typeⅠ(n= 196 strains)respectively.Conclusion The main epidemic clone of MRSA strain in this hospital is ST239-MRSA-SCCmec III-t030,antimicrobial resistance is serious,monitoring on drug-resistant strains in hospital should be strengthened.

Background and Objective:Thrombospondin-1(TSP1)is an inhibitor of angiogenesis and its promoter hypermethylation has been found resulting in gene silencing in some primary human carcinomas.This study was to investigate the promoter methylation of TSP1 and its correlation with TGF-β1 level and T cell lmmunity in gastric cardia adenocarcinoma(GCA).Methods:Methylation specific polymerase chain reaction(MSP)approach and immunohistochemistry method were used to examine the methylation status of the 5'CpG island and expression of TSP1 protein, respectively.The Ievel of TGF-β1 was measured by ELISA and T cell immunity of GCA by flow cytometry analysis.Results:TSP1 methylation frequency was significantly higher in tumor specimens than in corresponding normal tissues(35.4% vs.3.1%,P<0.001)and significanty higher in Stages Ⅲ and Ⅳ tumor tissues than in Stages Ⅰ and Ⅱ tumor tissues(P<0.05).TSP1 protein expression was significantly lower in the tumor tissues than in corresponding normal tissues (P<0.05)and statistically correlated with its methylation status(P0.05).The function of T cell immunity was significantly different between the GCA patients with hypermethylation of TSP1 and those without methylation of TSP1(P<0.05).Conclusion:Promoter hypermethylation of TSP1 may play an important role in the development of GCA and reflect the biological behaviours of GCA.