Adult ; Aged ; Aged, 80 and over ; Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism

Antibodies, Neutralizing ; Apoptosis ; Hep G2 Cells ; Humans ; Receptors, Tumor Necrosis Factor, Member 6b ; immunology ; metabolism

To investigate the effect of microRNA-221 (miR-221) on cell viability and apoptosis of hepatocellular carcinoma HepG2 cells. MiR-221 inhibitors and mimics were transfected into HepG2 cells. The expression of miR-221 was detected by real time quantitative RT-PCR. CellTiter-blue cell viability kit, Hoechst 33342/propidium iodide (PI) double staining assay, flow cytometry and Apo-ONE homogeneous caspase-3/7 kit were used to measure cell viability and apoptosis. MiR-221 inhibitors significantly inhibited the cell growth and miR-221 mimics increased cell viability 48 hours post-transfection measured by both CellTiter-blue cell viability kit and Hoechst 33342/PI assay (P is less than to 0.05). There was a positive correlation between these two assays (r = 0.993, P is less than to 0.01). With miR-221 mimics, the number of G1 stage cells (47.67%+/-1.53%) was significantly reduced as compared to the blank control (59.00%+/-1.00%) and the negative control (58.00%+/-1.00%, F = 81.77, P is less than to 0.01), and it was significantly raised in S stage (20.33%+/-1.15%) than in blank control (11.00%+/-1.00%) and negative control (12.00%+/-1.00%, F = 70.9, P is less than to 0.01) with flow cytometry analysis. More cell apoptosis and necrosis were significantly induced by miR-221 inhibitors 48 hours post-transfection detected by both Hoechst 33342/PI assay and flow cytometry PE Annexin V kit (P is less than to 0.05). The result from Apo-ONE homogeneous caspase-3/7 kit was consistent with the above two apoptotic assays, which showed that with miR-221 inhibitors, the activity of caspase-3/7 was significantly enhanced (P is less than to 0.05). MiR-221 contributes to the growth of hepatocellular carcinoma cells and miR-221 inhibition can induce cell apoptosis. miR-221 has the potential to become one of the new molecular targets for liver cancer therapy.
Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; Cell Proliferation ; drug effects ; Hep G2 Cells ; Humans ; Liver Neoplasms ; MicroRNAs ; metabolism

OBJECTIVETo study the expression of decoy receptor 3 (DcR3) and its relationship with apoptosis and prognosis in hepatocellular carcinoma (HCC).

METHODSThe expression of DcR3 protein in 43 cases of HCC and 16 cases of non-cancerous liver (including cirrhotic liver tissue and normal liver tissue adjacent to cavernous hemangioma) was studied by immunohistochemistry (using EnVision method). The status of apoptosis was evaluated by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) technique. Statistic analysis was carried out to assess the correlation between DcR3 expression, apoptotic index (AI) and clinicopathologic parameters.

RESULTSDcR3 protein was expressed in the cytoplasm of HCC cells. The positivity rate of DcR3 in HCC was 74.42% (32/43), which was significantly higher than that in the non-cancerous group (43.75%, P < 0.05). The positivity rate of DcR3 in HCC with metastasis detected within 20 months of diagnosis was 100% (22/22). This was significantly higher than that in HCC without metastasis (52.94%, P < 0.01). The DcR3 expression in HCC also correlated with serum alpha-fetoprotein level (r = 0.444, P < 0.01) and presence of tumor embolus in portal vein (r = 0.414, P < 0.01). However it had no relationship with the patient's age, sex, cirrhotic status, liver capsule invasion, number of tumor nodules and histologic differentiation (P > 0.05). The AI in HCC (0.78 +/- 0.64)% was significantly lower than that in the non-cancerous group [(3.32 +/- 1.81)%, P < 0.01]. The AI in clinical TNM stage I and II tumors (1.03 +/- 0.69)% was significantly higher than that in stage III and IV tumors [(0.52 +/- 0.48)%, P < 0.01]. The AI in HCC without metastasis (1.10 +/- 0.72)% was significantly higher than that in HCC without metastasis [(0.44 +/- 0.27)%, P < 0.05]. The AI correlated with serum alpha-fetoprotein level (r = -0.468, P < 0.01), presence of tumor embolus in portal vein (r = -0.434, P < 0.01) and liver capsule invasion (r = -0.331, P < 0.05). On the other hand, it had no relationship with patient's age, sex, cirrhotic status, number of tumor nodules and histologic differentiation (P > 0.05). The AI in DcR3-positive group (including both HCC and non-cancerous tissues) was significantly lower than that in DcR3-negative group (P < 0.01).

CONCLUSIONSThe expression of DcR3 in HCC correlates with apoptosis of tumor cells and may play a crucial role in tumor pathogenesis and progression. DcR3 protein expression and AI may also serve as important biologic indicators in predicting prognosis of HCC.

Adult ; Aged ; Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism ; physiology ; Retrospective Studies ; Young Adult

OBJECTIVETo observe the leg length discrepancy and accompanied knee varus/valgus deformity in matured patients with unilateral dislocation of the hip.

METHODSFrom March 2011 to December 2012, 28 patients who had unilateral dislocation of hip (Hartofilakidis classification II 17 cases and III 11 cases) were involved in this study.There were 6 male patients and 22 female patients, the age of the patients were 13.4-66.2 years, with mean age of 29.8 years. The standing anteroposterior full leg length X-ray films were obtained. Leg length discrepancy, the length of the femur, the length of the tibia and identified the varus/valgus knee deformities were measured. Statistical analysis was performed. A student's t test for paired samples was done for comparison of the parameters in the same patient between dislocated and undislocated leg, and the χ(2) test were used to assess valgus and varus knees, leg length discrepancy in high dislocation and low dislocation groups.

RESULTSSeventeen (60.7%) cases had longer femur length on the dislocated side than that on the undislocated side (t = 1.328, P = 0.197), with the maximum lengthening of 32.7 mm and a mean lengthening of 9.5 mm. Twenty-one (75.0%) cases had longer tibia length on the dislocated side (t = 3.039, P = 0.006), with a maximum lengthening of 10.9 mm and a mean lengthening of 4.5 mm. Twenty (71.4%) cases had longer relative leg length on the dislocated side (t = 2.451, P = 0.022), with a maximum lengthening of 25.0 mm and a mean lengthening of 9.4 mm. On the dislocated side of the leg, the degree of valgus angle was 3° ± 4°,while on the undislocated side, that was -3° ± 4°(t = 5.642, P = 0.000). On the dislocated side, 12 cases (42.9%) were of valgus deformities and 1 case was of varus deformity. On the contralateral side, 15 cases of varus deformities (53.6%) and 1 case of valgus deformity were observed(χ(2) = 18.139,P = 0.000).

CONCLUSIONSMost dislocated legs are longer in length than the contralateral side, both femur and tibia have also lengthened accordingly. Many knees on the dislocated side present valgus deformity, half of the knees on the contralateral side present varus deformity.

Adolescent ; Adult ; Aged ; Female ; Femur ; abnormalities ; diagnostic imaging ; Hip Dislocation, Congenital ; complications ; radiotherapy ; Humans ; Knee Joint ; abnormalities ; diagnostic imaging ; Leg Length Inequality ; diagnostic imaging ; etiology ; Male ; Middle Aged ; Radiography ; Tibia ; abnormalities ; diagnostic imaging ; Young Adult

OBJECTIVETo discuss the minimal invasive arthroscopic surgery technique and clinical results of both the medial and lateral meniscal transplantation following the anterior cruciate ligament reconstruction with double bundles and bone tunnels.

METHODSIn August 2008 a minimal invasive surgery of both the medial and lateral meniscal allograft transplantation following anterior cruciate ligament reconstruction was preformed for 1 case with both the medial and lateral meniscectomy by arthroscopic surgery. The method of two bone plugs attached on tibial plateau was employed for medial meniscal allograft transplantation and the technique the bridge in slot for lateral meniscal allograft transplantation. The VAS, Lysholm score and IKDC rating were recorded before and after operation. The stability of knee was assessed by Lachman test, drawer sign and pivot shift test.

RESULTSThe patient was followed up 26 month after the operations. The degrees of knee flexion, extension and function of walk were normal. The Lachman test, drawer sign and pivot shift test were nearly normal. The VAS after operation was 2 points lower than that before operation. The Lysholm score post-operation was 20 points higher than pre-operation. The IKDC became B degree in late following-up from C degree before the operation. MRI revealed anterior cruciate ligament graft was continuous and the meniscal allograft was normal shape on year 1 after the operation. The posterior horn of medial meniscal allograft and anterior corner of lateral meniscal allograft showed slightly shrunk. The second-look arthroscopy showed that the healing occurring between meniscal allograft and the capsule and meniscal allograft was normal shape on month 18 after the operation. The anterior horn of medial and lateral meniscus was slightly worn.

CONCLUSIONSBoth the medial and lateral meniscal transplantation following the anterior cruciate ligament reconstruction in appropriately selected patients with the medial and lateral meniscus-deficient knee may recover the knee mechanic balance and stability, which is a option of treatment for that young and activity patients. It is proposed that the medial and lateral meniscal grafts harvested from a single donator. Attention should be paid to the direction of the bone tunnels fixing the horns of the meniscus in order to avoid communication with the tunnels of anterior cruciate ligament reconstruction.

Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Reconstruction ; methods ; Arthroscopy ; Humans ; Knee Injuries ; surgery ; Male ; Menisci, Tibial ; transplantation ; Transplantation, Homologous ; Treatment Outcome ; Young Adult

BACKGROUNDRenal hypertrophy has been regarded as the early feature of diabetic nephropathy (DN), which may eventually lead to proteinuria and renal fibrosis. However, the exact mechanism of renal hypertrophy is still unclear. The aim of this study was to investigate the possible association of connective tissue growth factor (CTGF) with renal hypertrophy in uninephrectomized diabetic rats.

METHODSSeventy-two Sprague-Dawley (SD) rats were randomly divided into two groups: control group (group C, n = 32) and diabetic nephropathy (group DN, n = 40). Each group was re-divided into 4 subgroups according to the experimental period. The rats were sacrificed at 1, 2, 4, and 8 weeks respectively after induction of diabetes. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ) after rats had received uninephrectomy. Blood glucose (BG), body weight (BW), 24-h urinary albumin excretion (24hUalb), kidney weight (KW), KW/BW, glomerular tuft area (AG), glomerular tuft volume (VG), proximal tubular area (AT) at each time point, the width of glomerular basement membrane (GBM) and tubular basement membrane (TBM) at week 8 were measured when the rats were sacrificed. Renal expression of CTGF and p27kip1 were detected by immunohistochemical staining. The relationship between CTGF expression and increasing of VG and AT was analyzed.

RESULTSThere was a significant increase of 24hUalb, KW, and KW/BW from week 1 onward in diabetic rats compared to those in group C (P < 0.05, respectively), diabetic rats also had a significant increase of AG, VG, and AT from week 1 onward. It was also shown that diabetic rats had a thickening of GBM [(245.7 +/- 103.0) nm vs (121.8 +/- 19.1) nm, P < 0.01] and TBM [(767.7 +/- 331.1) nm vs (293.0 +/- 110.5) nm, P < 0.01] at week 8. There was a weak expression for CTGF and p27kip1 in normal glomeruli and tubuli, while a significant increasing expression of CTGF and p27kip1 was found in glomeruli and tubuli in diabetic kidney from week 1 onward (P < 0.05, respectively), and the extent of CTGF expression was positively correlated with AG (r = 0.92, P < 0.05), VG (r = 0.86, P < 0.05), AT (r = 0.94, P < 0.01) and positively correlated with the expression of p27kip1 (r = 0.96, P < 0.01).

CONCLUSIONThe expression of CTGF increases in diabetic rat kidney at the early stage, which might be an important mediator of renal hypertrophy through arresting cell cycling.

Albuminuria ; etiology ; Animals ; Connective Tissue Growth Factor ; Cyclin-Dependent Kinase Inhibitor p27 ; analysis ; Diabetes Mellitus, Experimental ; pathology ; Hypertrophy ; Immediate-Early Proteins ; analysis ; physiology ; Intercellular Signaling Peptides and Proteins ; analysis ; physiology ; Kidney ; pathology ; Male ; Nephrectomy ; Rats ; Rats, Sprague-Dawley ; Streptozocin

Since the research of molecular identification of Chinese Materia Medica (CMM) using DNA barcode is rapidly developing and popularizing, the principle of this method is approved to be listed in the Supplement of the Pharmacopoeia of the People's Republic of China. Based on the study on comprehensive samples, the DNA barcoding systems have been established to identify CMM, i.e. ITS2 as a core barcode and psbA-trnH as a complementary locus for identification of planta medica, and COI as a core barcode and ITS2 as a complementary locus for identification of animal medica. This article introduced the principle of molecular identification of CMM using DNA barcoding and its drafting instructions. Furthermore, its application perspective was discussed.
Animals ; China ; DNA ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; classification ; isolation & purification ; Electron Transport Complex IV ; genetics ; Materia Medica ; classification ; isolation & purification ; Medicine, Chinese Traditional ; Plant Proteins ; genetics ; Plants, Medicinal

OBJECTIVETo discuss the clinical features and surgical treatments of giant coronary artery aneurysm (CAA).

METHODSFrom July 1996 to October 2004, 6 giant CAA patients were underwent surgery at Fuwai hospital. Three cases were underwent CAA resection, 2 concomitant coronary bypass, 3 reconstruction. The giant CAA was often combined with other cardiac diseases. Four cases underwent additional procedures of fistula closure, 3 aortic valve replacements, 2 aortoplasty and 1 thrombus cleaning at the same time.

RESULTSAll patients recovered uneventfully. The mean of cardiopulmonary bypass time was (144 +/- 26) min (range 67 to 207 min). Aortic cross clamping time was (104 +/- 21) min (range 56 to 172 min). Patients follow-up time occurred from 8 to 87 months (mean of 48 months). All patients were free of symptoms during follow-up. None of the patients died during the follow-up period and none of the CAA recurred.

CONCLUSIONSThe giant CAA is a serious cardiovascular disease, early diagnosis and surgical treatment are mandatory.

Adult ; Coronary Aneurysm ; pathology ; surgery ; Coronary Artery Bypass ; Coronary Vessels ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Surgical Procedures, Operative ; methods ; Treatment Outcome

Fingerprints of lipophilic components in the roots of Salvia miltiorrhiza and S.yunnanensis were analyzed by UPLC-DADand UPLC coupled with mass spectroscopy to evaluate the differences and similarities of the lipophilic components in the two kinds of herbs.The UPLC analysis of 18 batches of S.miltiorrhiza and 16 batches of S.yunnanensis was performed on a 25℃Thermo Accucore C_(18)column(2.1 mm×100 mm,2.6μm)by Shimadzu LC-20AD;mobile phase was 0.026%phosphoric acid(A)-acetonitrile(B)with gradient elution;flow rate was 0.4 m L·min~(-1);detection wavelength was set at 270 nm;injection volume was 2μL.The molecular structures of the lipophilic components were analyzed on a 25℃Thermo Accucore C_(18)column(2.1 mm×100 mm,2.6μm)by Thermo U3000 UPLC Q Exactive Orbitrap LC-MS/MS with a mobile phaseconsisting of 0.1%formic acid water(A)and 0.1%formic acidacetonitrile(B).The mass spectrometry was acquired in positive modes using ESI.There are 10 common peaks in the lipophilic components of S.miltiorrhiza.The similarity between the 16 batches of S.miltiorrhiza and their own reference spectra was greater than 0.942,and the average similarity was 0.973.There are 12 common peaks in the lipophilic components of S.yunnanensis.The similarity between the 18 batches of S.yunnanensis and their own reference spectra was greater than 0.937,and the average similarity was 0.976.The similarity between the reference chromatograms of S.miltiorrhiza and S.yunnanensis was only 0.900.There are three lipophilic components in S.yunnanensis,which are not found in S.miltiorrhiza,and one of which isα-lapachone.There is a lipophilic component in S.miltiorrhiza not found in S.yunnanensis,which may be miltirone.The two herbs contain 8 common lipophilic components including dihydrotanshinoneⅠ,cryptotanshinone,tanshinoneⅠ,tanshinoneⅡ_A,nortanshinone in which the content of tanshinoneⅡ_A,dihydrotanshinoneⅠand cryptotanshinone of S.yunnanensisis significantly lower than that of S.miltiorrhiza(P<0.01),and the contents of tanshinoneⅠand nortanshinone are significantly lower than that of S.miltiorrhiza too(P<0.05).There are significant differences in the types and contents of lipophilic components between the roots of S.miltiorrhiza and S.yunnanensis,and the similarity between the fingerprints of interspecies is much lower than that between the same species.Therefore,the roots of S.miltiorrhiza and S.yunnanensis are two kinds of herbs which are quite different in chemical compounds and compositions.
Chromatography, Liquid ; Diterpenes, Abietane ; Molecular Structure ; Plant Roots ; Salvia miltiorrhiza ; Tandem Mass Spectrometry