Objective To lay the foundation for analyzing the mechanism of liver cell injury caused by mtDNA deletion and mutation in patients with obstructive jaundice. Methods 30 patients were randomly selected as obstructive jaundice group (case group) and 10 patients as control group according to the strict condition. Author makes use of the methods of PCR amplification of the entire human mitochondrial genome in 17 mismatch-specific overlapping fragments and gene sequencing results to Preliminary estimate the localizathion of hepatocyte mtDNA damage in patients with obstructive jaundice. Result Deletions and length of partial liver cells were 8429-9591 of about 1. 1 kb, 16024-60 of about 0. 6 kb, 1889-3031 of about 1. 1 kb and 4977bps common deletion and the high mutation rate of some bases in D-loop region. Conclusion There are multiple mtDNA deletions and multiple point mutations in patients with obstructive jaundice

Adult ; China ; epidemiology ; Extraction and Processing Industry ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Occupational Exposure ; Pneumoconiosis ; epidemiology ; Young Adult

Objective To evaluate the role of prostaglandin E2 (EP) receptors in H9c2 cardiomyocyte hypertrophy induced by prostaglandin E2 (PGE2).Methods Primary cultured H9c2 cardiomyocytes were seeded in culture flasks (3 ml/flask) or in 24-well plate (1 ml/hole) or 6-well plate (2 ml/hole) with density of 4 × 104/ml.The cells were randomly divided into 4 groups (n=24 each): control group (group C),PGE2 group,AH6809 (EP1 and EP2 receptor antagonist) group (group A) and GW627368X (EP4 receptor antagonist) group (group G).The cells were continuously cultured for 48 h.PGE2 (final concentration 1 μmol/L) was added to the culture medium in PGE2 group.PGE2 (final concentration 1 μmol/L) and A H6809 (final concentration 10 μmol/L) were added to the culture medium in group A.PGE2 (final concentration 1 μmol/L) and GW627368X (final concentration 10 μmol/L) were added to the culture medium.The cells were then cultured for 48 h in groups PGE2,A and G.Then the cell morphology was observed by using fluorescent microscope.The cell diameter was measured by using the Image J medical image analysis system.Total protein content in the cells was measured with BCA method.The expression of atrial natriuretic peptide (ANP) mRNA and brain natriuretic peptide (BNP) mRNA in the cytoplasm was determined using RT-PCR.Results Compared with group C,the total protein in the cells and cell diameter were significantly increased,and the expression of ANP mRNA and BNP mRNA in the cytoplasm was up-regulated in groups PGE2,A and G (P ＜ 0.05).Compared with group PGE2,the total protein in the cells and cell diameter were significantly decreased,and the expression of ANP mRNA and BNP mRNA in the cytoplasm was downregulated in group G (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group A (P ＞ 0.05).Conclusion EP4 receptor mediates H9c2 cardiomyocyte hypertrophy induced by PGE2 and the effect is not related to EP1 and EP2.

Objective:To observe the effects of lentivirus-mediated shRNA silencing of PRL-3 gene on the proliferation, migration and invasion of lung cancer A549 cells and regulation of epithelial-mesenchymal transition (EMT) signaling pathway.Methods:Lung cancer A549 cells were transfected with lentiviral interference vector carrying PRL-3 shRNA to build a stable PRL-3-silencing cell line. The cells were divided into blank control group, NC shRNA group (negative control group) and PRL-3 shRNA group (PRL-3 inhibiting RNAi lentivirus group). CCK-8 method, colony formation assay, Transwell and invasion chamber assay were performed to detect the proliferation, migration and invasion ability of A549 cells respectively. The expressions of E-cadherin and Snail mRNA were detected by real-time fluorescent quantitative PCR.Results:The stable PRL-3-silencing cell line was successfully constructed. The knockdown efficiency of PRL-3 gene in the PRL-3 shRNA group reached 83.5%. CCK-8 method detected the proliferation ability of A549 cells, and the results showed the 24 h absorbance ( A) values of A549 cells in the blank control group, NC shRNA group and PRL-3 shRNA group were 0.296±0.008, 0.342±0.007 and 0.292±0.004, with a statistically significant diffe-rence ( F=106.300, P<0.001), and the PRL-3 shRNA group was significantly lower than the NC shRNA group ( P<0.001); at 48, 72, 96 h after transfection, the cell proliferation abilities of the PRL-3 shRNA group were also significantly inhibited. Colony formation assay showed that the numbers of colony formation in the blank control group, NC shRNA group and PRL-3 shRNA group were 166.7± 6.7, 158.0±6.1 and 119.7±1.5 ( F=67.290, P<0.001). The ability of colony formation of the PRL-3 shRNA group was significantly lower than that of the NC shRNA group ( P<0.001). The numbers of migrated cells in the blank control group, NC shRNA group and PRL-3 shRNA group were 100.0±1.9, 98.8±1.9 and 44.6±7.6 ( F=430.300, P<0.001). The migration ability of the PRL-3 shRNA group was significantly lower than that of the NC shRNA group ( P<0.001). The numbers of invaded cells in the three groups were 117.7±4.1, 113.1±6.6 and 55.6±8.4 ( F=247.200, P<0.001). The invasion ability of the PRL-3shRNA group was significantly lower than that of the NC shRNA group ( P<0.001). Real-time fluorescent quantitative PCR detection results showed that after silencing the expression of PRL-3, the relative expression level of E-cadherin mRNA in A549 cells was significantly up-regulated, and the level of Snail mRNA was significantly down-regulated. Conclusion:PRL-3 silencing can inhibit the proliferation, migration and invasion of A549 cells effectively. PRL-3 may affect the invasion of lung cancer cells through the EMT pathway.

Objective: To explore the mechanism of An-pressing manipulation in improving post-stroke muscle spasticity, by observing the changes of γ-aminobutyric acid (GABA) and glycine (Gly) in plasma and gray matter of L1-L3 spinal cord anterior horn in post-stroke rats with muscle spasticity after An-pressing manipulation intervention. Methods: Ten of 80 adult male Sprague-Dawley (SD) rats were randomly selected as the blank group, and the remaining 70 were used for modeling. The middle cerebral artery occlusion (MCAO) rat model was established by insertion suture occlusion method in the left external carotid artery. Thirty rats with a Longa neurological score of 2-3 points and a modified Ashworth spasticity scale score of 1-, 1+, or 2 were included in the experiment. Using the random number table method, the 30 successfully modeled rats were randomly divided into a model group, an An-pressing tendon group and an An-pressing muscle belly group. Two days after modeling, rats in the An-pressing tendon group and An-pressing muscle belly group received An-pressing manipulation on the tendon and belly of quadriceps femoris muscle respectively, with the pressure of (350±50) g and the frequency of 5 s/time, 15 min per session, once a day for 5 continuous days. After the 5th treatment, the tension of the rat quadriceps femoris muscle was evaluated using the modified Ashworth spasticity scale. The Gly levels in rat plasma and L1-L3 segments of spinal cord were determined by enzyme-linked immunosorbent assay (ELISA). The GABA levels in rat plasma and L1-L3 segments of spinal cord were measured by high performance liquid chromatography (HPLC). Results: The decrease in rat muscle tension scored by the modified Ashworth spasticity scale in the An-pressing tendon group was more significant than that in the An-pressing muscle belly group (P<0.01); the increases in Gly and GABA levels in the rat plasma and L1-L3 segments of spinal cord were more significant in the An-pressing tendon group than those in the An-pressing muscle belly group (all P<0.01). Conclusion: Based on the theory of 'anti-stretch reflex' of tendon organs, the use of An-pressing manipulation to induce the 'anti-stretch reflex' by stimulating the tendon organs can improve the muscle spasticity of rats, which is better than An-pressing the muscle belly. Increased levels of Gly and GABA in rat plasma and L1-L3 segments of spinalcord may be one mechanism of An-pressing manipulation to improve muscle spasticity by stimulating tendon organs.

Adenocarcinoma ; diagnosis ; pathology ; Adult ; Aged ; Aged, 80 and over ; Ascitic Fluid ; chemistry ; Cadherins ; analysis ; Calbindin 2 ; Carcinoembryonic Antigen ; analysis ; Diagnosis, Differential ; Epithelial Cells ; chemistry ; Female ; Humans ; Immunohistochemistry ; Keratin-5 ; analysis ; Male ; Middle Aged ; Pericardial Effusion ; chemistry ; diagnosis ; Pleural Effusion ; chemistry ; diagnosis ; Pleural Effusion, Malignant ; chemistry ; diagnosis ; S100 Calcium Binding Protein G ; analysis

Objective To investigate the effects of different modes of one-lung ventilation (OLV) on hemodynamies in the patients undergoing thoracic operation.Methods Forty-five adult patients undergoing thoracic surgery,were randomly allocated into 3 groups based on the modes of OLV used ( n =15 each):intermittent positive pressure ventilation ( IPPV,VT 6-8 ml/kg,RR 10-14 bpm,I:E 1:2) group,IPPV + positive end-expiratory pressure (PEEP) group and IPPV + continuous positive airway pressure (CPAP) group.Double-lumen tube was inserted.Conrrect positioning was verified by fiberoptic bronchoscopy.The patients were mechanically ventilated.PETCO2 was maintained at 35-45 mm Hg.In group IPPV + PEEP,OLV was performed for 30 min with PEEP of 5 cm H2O and then for another 30 min with PEEP of 10 cm H2O.In group IPPV + CPAP,OLV was performed with IPPV in the lung on the ventilated side and with CPAP of 5 cm H2O in the lung on the operated side (for 1 h).MAP,HR,cardiac output (CO),cardiac index ( CI),stroke volume (SV),and stnoke volume index (SVI) were recorded before induction of anesthesia,at 10 min after intubation,at 30 min of two-lung ventilation,at 30 min and 1 h of OLV,and at the end of operation ( T1-6 ).Arterial blood samples were taken at T1,2,4-6 for blood gas analysis.The levels of blood glucose and lactate were measured.Oxygen delivery ( DO2 ) and DO2 index ( DO2I) were calculated.Results Compared with IPPV group,SV,SVI,CO,CI,DO2 and DO2I were significantly decreased at T4,5 ( P ＜ 0.05),and no significant change was found in the levels of blood glucose and lactate in group IPPV + PEEP,and no significant change was found in the parameters mentioned above in group IPPV + CPAP ( P ＞ 0.05).Compared with IPPV + PEEP group,SV,SVI,CO,CI,DO2 and DO2I were significantly increased at T4,5 ( P ＜ 0.05),and no significant change was found in the levels of blood glucose and lactate in group IPPV + CPAP ( P ＞ 0.05).Conclusion It exerts no influence on hemodynamics using OLV with IPPV in the lung on the ventilated side and with CPAP of 5 cm H2O in the lung on the operated side,however,OLV with IPPV + PEEP can result in hemodynamic fluctuation,but the degree of fluctuation is lesser and DO2 can be maintained in the patients undergoing thoracic operation.

OBJECTIVETo study the effects of Chinese medicine tetramethylpyrazine (TMP) on intracellular free calcium ([Ca2+]i) in cultured penis corpus cavernosum smooth muscle cell (PCSMC) in rabbits.

METHODSBy using laser scanning confocal microscope (LSCM), the [Ca2+]i fluorescence signal changes was investigated in cultured PCSMC loaded with Ca2+ indicator Fluo-3/AM and divided into potassium chloride(KCl) group and norepinephrine (NE) group. Compared with verapamil (Ver), the effects of TMP was observed in different concentrations on [Ca2+]i increase induced by high potassium and NE.

RESULTSTMP had no obvious effect on resting PCSMC [Ca2+]i. It was found that 1, 10, 100 mumol/L TMP significantly inhibited [Ca2+]i increase induced by high potassium-depolarization. The peak inhibition rates were (38.6 +/- 3.0)%, (44.1 +/- 2.4)% and (53.7 +/- 4.1)% respectively. TMP could also inhibit cytosolic calcium pool release induced by 1 mumol/L NE. The peak inhibition rates were (13.9 +/- 2.7)%, (21.2 +/- 1.9)% and (29.5 +/- 3.6)% respectively.

CONCLUSIONSTMP can inhibit rabbit PCSMC [Ca2+]i significantly by suppressing voltage-dependent calcium channel and cytosolic calcium pool release. The effect, similar to Ver, signifies the important mechanism of erectile dysfunction (ED) therapy.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Dose-Response Relationship, Drug ; Male ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Penis ; blood supply ; metabolism ; Pyrazines ; pharmacology ; Rabbits ; Vasodilator Agents ; pharmacology

OBJECTIVETo study clinical outcomes of Chinese medidine fumigation and massage therapy for the treatment of knee stability and functional recovery after anterior cruciate ligament reconstruction operation,and to explore the effect on tendon-bone healing.

METHODSTotal 50 patients were divided into two groups: the control group (normal rehabilitation therapy group),the treatment group (Chinese medicine fumigation and manipulation group). There were 25 patients in the control group, including 16 males and 9 females, who were treated with isometric muscle training, with the gradually enlarging amplitude of flexion and progressive loading of bearing training for knee recovery. There were 25 patients in the treatment group, including 15 males and 10 females,who were treated with the conventional rehabilitation therapy combined with Chinese medicine fumigation and massage therapy. The Chinese herbs named as Haitongpi decoction was steamed by a special equipment to fumigate the knee after operation; Based on the biomechanical parameters of the ligament reconstruction, the massage therapy was designed to control the degree of the knee flexion and release the adhesion for early recovery of knee functions. The Lysholm knee function evaluation system was used, and MRI examination was performed to measure the change in width of ligament tunnel in femur and tibia to evaluate the safety and stability of the treatment.

RESULTSLysholm system showed that two groups both had improving results from the 1st month after operation to the 3rd month (treatment group, F=36.54, P<0.05; the control group, F=28.12, P<0.05), and the results of the treatment group was better than that of the control group at the observation point (the 1st month, t=0.105, P<0.05; the 3rd month, t=5.361, P<0.01). There was no difference between the two groups when evaluating the bone and tendon healing 3 and 12 months after operation (P>0.05), indicating that Chinese rehabilitation therapy was a safety treatment without the influence on the loosing of tendon.

CONCLUSIONChinese medicine fumigation and massage therapy can early improve the knee function after the anterior cruciate ligament reconstruction operation without the disturbance of the knee stability.

Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Injuries ; Anterior Cruciate Ligament Reconstruction ; Case-Control Studies ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Fumigation ; Humans ; Knee Injuries ; drug therapy ; physiopathology ; surgery ; therapy ; Knee Joint ; drug effects ; physiopathology ; surgery ; Male ; Massage ; Range of Motion, Articular ; Recovery of Function

OBJECTIVETo discuss the feasibility of vascular bundle implantation combined with allogeneic bone marrow stromal cells (BMSCs) transplantation in treating rabbit femoral head osteonecrosis and bone defect, in order to explore a new method for the treatment of femoral head necrosis.

METHODSThirty-six New Zealand rabbits were randomly divided into three groups,with 12 rabbits in each group. Bilateral femoral heads of the rabbits were studied in the experiment. The models were made by liquid nitrogen frozen, and the femoral heads were drilled to cause bone defect. Group A was the control group,group B was stem cells transplantaion group of allograft marrow stromal,and group C was stem cells transplantation group of allograft marrow stromal combined with vascular bundle implantation. Three rabbits of each group were sacrificed respectively at 2, 4, 8, 12 weeks after operation. All specimens of the femoral heads were sliced for HE staining. Furthermore ,vascular density and the percentage of new bone trabecula of femoral head coronary section in defect area were measured and analyzed statistically.

RESULTSIn group C,new bone trabecula and original micrangium formed at the 2nd week after operation; new bone trabecula was lamellar and interlaced with abundant micrangium at the 8th week;at the 12th week,the broadened,coarsened bone trabecula lined up regularly,and the mature bone trabecula and new marrow were visible. At the 2nd week after operation,there was no statistical significance in the percentage of new bone trabecula of femoral head coronary section in defect area between group B and C. While at 4, 8, 12 week after operation, vascular density and the percentage of new bone trabecula of femoral head coronary section in defect area of group C was higher than that of group B.

CONCLUSIONAllogeneic bone marrow stromal cells cultured in vivo can form new bone trabecula, and can be applied to allotransplant. Vascular bundle implanted into the bone defect area of femoral head necrosis could improve blood supply, and promote the formation of bone trabecula.

Animals ; Blood Vessels ; transplantation ; Combined Modality Therapy ; Female ; Femur Head Necrosis ; pathology ; surgery ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; Rabbits ; Transplantation, Homologous