This research is to study the relationship between HPLC fingerprints of Moutan Cortex, Paeoniae Radix Rubra and Paeoniae Radix Alba and their activity on lipopolysaccharide-induced acute lung injury. HPLC fingerprints of each extract of Moutan Cortex,Paeoniae Radix Rubra and Paeoniae Radix Alba were established by an optimized HPLC-MS method. The activities of all samples against protein and tumor necrosis a factor were tested by the model of lipopolysaccharide-induced acute lung injury. The possible relationship between HPLC-MS fingerprints and the activitieswere deduced by the Partial least squares regression analysis method. Samples were analyzed by HPLC-MS/MS to identify the major peaks. The results showed that each sample had some effect on acute lung injury. Four components with a lager contribution rate of efficacy were calculated by the research of spectrum-effect relationship. Moutan Cortex exhibited good activity on acute lung injury, and gallic acid, paeoniflorin, galloylpaeoniflorin and paeonol were the main effective components.
Acetophenones ; chemistry ; pharmacology ; Acute Lung Injury ; chemically induced ; drug therapy ; Animals ; Bridged Bicyclo Compounds, Heterocyclic ; chemistry ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gallic Acid ; chemistry ; pharmacology ; Glucosides ; chemistry ; pharmacology ; Lipopolysaccharides ; pharmacology ; Male ; Monoterpenes ; chemistry ; pharmacology ; Paeonia ; chemistry ; Plant Roots ; chemistry ; Rats ; Rats, Wistar ; Tandem Mass Spectrometry ; methods

OBJECTIVETo study the effect of osteopontin (OPN) expression down-regulated by RNA interference (RNAi) on the invasiveness of hepatocelluar carcinoma cell line HCC-LM3.

METHODSHCC-LM3 cells were transfected with the chemically synthesized small interfering RNA (siRNA) formulated by lipofectamine 2000. Wild type HCC-LM3 and HCC-LM3 cells transfected with non-specific siRNA served as controls. Real-time PCR and Western blotting were used to quantify the mRNA and OPN protein levels. The malignant phenotypes of transfected HCC-LM3 cells including cellular growth rate, colony formation and Matrigel invasion activities were analyzed.

RESULTSSequence-specific siRNAs targeting OPN suppressed OPN RNA expression by 79% and also decreased OPN protein level by 81% in HCC-LM3 cells. The number of formed colonies and migrating numbers in vitro were decreased in HCC-LM3 cells transfected using sequence-specific siRNAs targeting OPN relative to controls (P < 0.05).

CONCLUSIONThis study demonstrated that specific siRNA is able to reduce OPN at both the mRNA and protein levels and significantly diminishes the invasiveness of hepatocellular carcinoma cells.

Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Down-Regulation ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Invasiveness ; Osteopontin ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Transfection

Adolescent ; Adult ; Aged ; Female ; Humans ; Laparoscopy ; Male ; Middle Aged ; Splenectomy ; methods

Our previous studies showed that spinal neurons sensitization was involved in morphine withdrawal response. This study was to investigate the roles of spinal protein kinase C (PKC) alpha, gamma in morphine dependence and naloxone-precipitated withdrawal response. To set up morphine dependence model, rats were subcutaneously injected with morphine (twice a day, for 5 d). The dose of morphine was 10 mg/kg in the first day and was increased by 10 mg/kg each day. On day 6, 4 h after the injection of morphine (50 mg/kg), morphine withdrawal syndrome was precipitated by an injection of naloxone (4 mg/kg, i.p.). Chelerythrine chloride (CHE), a PKC inhibitor, was intrathecally injected 30 min before the administration of naloxone. The scores of morphine withdrawal symptom and morphine withdrawal-induced allodynia were observed. One hour after naloxone-precipitated withdrawal, Fos protein expression was assessed by immunohistochemical analysis and Western blot was used to detect the expression of cytosol and membrane fraction of PKC alpha and gamma in the rat spinal cord. The results showed that intrathecal administration of CHE decreased the scores of morphine withdrawal, attenuated morphine withdrawal-induced allodynia and also inhibited the increase of Fos protein expression in the spinal cord of morphine withdrawal rats. The expression of cytosol and membrane fraction of PKC alpha was significantly increased in the spinal cord of rats with morphine dependence. Naloxone-precipitated withdrawal induced PKC alpha translocation from cytosol to membrane fraction, which was prevented by intrathecal administration of CHE. During morphine dependence, but not naloxone-precipitated withdrawal, PKC gamma in the spinal cord translocated from cytosol to membrane fraction, and intrathecal administration of CHE did not change the expression of PKC gamma in the spinal cord of naloxone-precipitated withdrawal rats. It is suggested that up-regulation and translocation of PKC in the spinal cord contribute to morphine dependence and naloxone-precipitated withdrawal in rats and that PKC alpha and gamma play different roles in the above-mentioned effect.
Animals ; Male ; Morphine Dependence ; physiopathology ; Naloxone ; pharmacology ; Protein Kinase C ; metabolism ; physiology ; Protein Kinase C-alpha ; metabolism ; physiology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; metabolism ; physiopathology ; Substance Withdrawal Syndrome ; enzymology ; physiopathology

OBJECTIVETo summarize the experience of laparoscopic common bile duct exploration.

METHODSThe clinical data of 587 cases who underwent laparoscopic common bile duct exploration from June 1992 to May 2006 were analyzed.

RESULTSThe surgery was successful in 585 cases (99.7%), 2 cases were converted to open common bile duct exploration. The duration of operation was 60 approximately 230 min (averaged 85 min), the complications consisted of biliary fistula (n=13), injury of the duodenum (n=1), abscess of drainage tube orifice (n=1), titanium clip discharging out from T tube (n=3), residual common bile duct stones (n=35). The patients could take food and walk on the second postoperative day and average postoperative hospital stay was 4.6 days.

CONCLUSIONSLaparoscopic common bile duct exploration is a safe and effective procedure in treating the calculus of bile duct.

Adult ; Aged ; Aged, 80 and over ; Biliary Tract Diseases ; surgery ; Biliary Tract Surgical Procedures ; methods ; Common Bile Duct ; surgery ; Female ; Humans ; Laparoscopy ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Retrospective Studies ; Treatment Outcome

Objective To assess the effect of preoperative administration of tranexamic acid (TXA) on the hidden blood loss in the surgery of intertrochanteric fractures with proximal femoral nail anti-rotation (PFNA).Methods Eighty patients with intertrochanteric fracture were treated with PFNA in our hospital from November 2015 to July 2016.They were 15 men and 65 women,with a mean age of 72.6 years.Of them,39 were included into TXA group where TXA was administered preoperatively and 41 were assigned into the control group where no TXA was used preoperatively.Blood routine examinations were carried out on one day before operation,the first and third days after operation.The surgical blood loss,operative blood transfusion,24-hour drainage after operation,and postoperative 3-day blood transfusion were recorded.The total and hidden blood losses were calculated according to the formula.The 2 groups were compared in terms of blood loss and complications.Results In TXA group,the total blood loss (1,632.3 ± 849.2 mL),the hidden blood loss (1,270.9 ± 623.3 mL) and the transfusion rate (28.2％) were significantly lower than those in the control group (2,014.8 ± 924.7 mL,1,549.1 ± 624.9 mL and 56.1％) (P ＜ 0.05).There was no significant difference between the 2 groups in visible blood loss (361.4 ± 154.3 mL for TXA group versus 465.7 ± 191.3 mL for the control group) (P ＞ 0.05).Deep venous thrombosis occurred in 2 patients,limb swelling in one patient and wound dehiscence in one patient in TXA group while deep venous thrombosis occurred in one patient,limb swelling in 3 patients and wound dehiscence in none in the control group,showing no significant differences between the 2 groups (P ＞ 0.05).No infection was observed in either group.Conclusion Preoperative administration of TXA can reduce surgical hidden blood loss and transfusion rate as well but will not increase the risk of deep vein thrombosis in the surgery of intertrochanteric fractures with PFNA.

OBJECTIVETo observe the therapeutic effect of tranquilizing and allaying excitement needling method on insomnia of heart and spleen deficiency and the effect of brain blood flow.

METHODSSixty cases were randomly divided into a tranquilizing and allaying excitement needling method group (observation group) and an eight confluence points selected group (control group), 30 cases in each group. The observation group was treated by acupuncture at Sishencong (EX-HN 1), Shenmen (HT 7), and Sanyinjiao (SP 6) with tranquilizing and allaying excitement needling method. The control group was treated by acupuncture at Shenmai (BL 62) and Zhaohai (KI 6). Their therapeutic effects and changes of brain blood flow were observed.

RESULTSThe total effective rate was 93.3% (28/30) in the observation group which was better than 83.3% (25/30) in the control group (P < 0.05). After treatment, the peak velocity of systolic (Vp) and diastolic blood flow velocity (Vd) of middle cerebral artery, basilar artery and vertebral artery were increased in the both groups (P < 0.01, P < 0.05), with more obvious increase in the observation group (P < 0.01, P < 0.05), except the Vd of left vertebral artery.

CONCLUSIONBoth the tranquilizing and allaying excitement needling method and the eight confluence points selected needling method can improve the clinical signs and symptoms. Meanwhile, the therapeutic effect of acupuncture is related with improvement of brain blood flow. However, the tranquilizing and allaying excitement needling method has better therapeutic effect on insomnia of heart and spleen deficiency.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Brain ; blood supply ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Sleep Initiation and Maintenance Disorders ; physiopathology ; therapy ; Spleen ; physiopathology ; Young Adult

OBJECTIVETo investigate the expression of PTEN (phosphatase and tension homology deletion on chromosome 10, PTEN) and its pseudogene PTENP1 in acute leukemia (AL) and correlation between them, and to explore the role of PTENP1 on the PTEN expression in AL cells.

METHODSPTEN and PTENP1 mRNA expression were evaluated in bone marrow (BM) samples from 138 newly diagnosed AL patients and 15 healthy controls by quantitative real-time RT-PCR (qRT-PCR). pCDH1-PTENP1 3'UTR-GFP lentivirus vectors were constructed. 293T cells were transfected by calcium phosphate precipitation to produce retrovirus. HL-60 cell line was infected with the retroviral vectors expressing pCDH1-GFP and pCDH1-PTENP1 3'UTR-GFP respectively. The flow cell sorter was used to sort the HL-60 with GFP positively expressed. The mRNA expression of PTEN and PTENP1 was detected by qRT-PCR, the expression of PTEN protein by western blot, and the impact of PTENP13'UTR on the proliferation of HL-60 cells by MTT assay.

RESULTSAML patients showed significantly lower PTEN and PTENP1 mRNA expression in BM compared to healthy controls. Correlation analysis showed that the expression of PTEN and PTENP1 mRNA were positively correlated (P < 0.05). The 108 cases of PTENP1(+) AML were classified according to the prognostic classification of 2011 NCCN Clinical Practice Guidelines in AML, there was no difference among different subgroups. HL-60 cell line was infected with the retroviral vectors expressing pCDH1-GFP (control group) and pCDH1-PTENP1 3'UTR-GFP respectively. Compared with the control group, PTENP1 mRNA level of HL-60 infected with the retroviral vectors expressing pCDH1-PTENP1 3'UTR-GFP increased significantly, and PTEN mRNA level also increased. While the PTEN protein level and the cell growth rate of the PTENP1 3'UTR group didn't change significantly.

CONCLUSIONPTEN and PTENP1 mRNA expression level of BM cells from AL patients is significantly lower. There is a positive correlation between expression of PTEN and PTENP1 mRNA. PTENP1 may regulate the expression of PTEN in mRNA level.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Female ; Gene Expression ; HL-60 Cells ; Humans ; Leukemia ; genetics ; Male ; Middle Aged ; PTEN Phosphohydrolase ; genetics ; Pseudogenes ; genetics ; RNA, Messenger ; genetics ; Transfection ; Young Adult

AIMTo study the effect of thyroid hormone on protein kinase C activity and isoprotein expressions in cardiac myocytes and fibroblasts of rats in vitro.

METHODSCardiac myocytes and fibroblasts were cultured according to the method of Simpson. Cells were pretreated with 1% newborn calf serum (NCS) or Angiotensin II (Ang II) for 24 hours, then Triiodothyronine (T3) was added to the culture medium and the culture was kept for another 48 hours. The protein kinase C activation were measured by PepTaga non-radioactive PKC assay, and the expressions of PKC alpha and PKC epsilon were detected by Western blot method.

RESULTSAt the condition of 1% NCS culture medium, T3 could inhibit PKC activity and PKC epsilon expression in cardiac myocytes significantly, but the expression of PKC alpha in cardiac myocytes was not influenced by T3. In cardiac fibroblasts, neither PKC activity nor PKC alpha and PKC epsilon expressions was influenced by T3. When cells were pretreated with Ang II for 24 hours, PKC activities in cardiac myocytes and fibroblasts were increased significantly, and PKC epsilon expressions in cardiac myocytes were also markedly increased. Following a T3 treatment, PKC activity and PKC epsilon expression in cardiac myocytes were markedly decreased, but PKC activity in cardiac fibroblasts was not changed.

CONCLUSIONWhether at the condition of 1% NCS medium or in a pretreatment with Ang II, thyroid hormone could inhibit the PKC activity and PKC epsilon expression in cardiac myocytes. The influence of thyroid hormone on the PKC signal pathway in cardiac myocyte may be involved in many pathophysiological progress of myocardium.

Animals ; Cells, Cultured ; Myoblasts, Cardiac ; drug effects ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; Protein Kinase C ; metabolism ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Thyroid Hormones ; pharmacology

BACKGROUNDEndometrial polyps (EPs) occur in approximately 34.9% of infertile women. Transvaginal sonography (TVS) is a routine, non-invasive component of fertility evaluation. Most ultrasonographic studies of EPs have focused on abnormal uterine bleeding; few have assessed EPs in infertile women. Furthermore, no studies have explored endometrial thickness and its correlation with EPs in infertile women. This study aimed to assess transvaginal sonographic assessment of endometrial thickness and its value in diagnosis and prediction of EPs in infertile women.

METHODSA retrospective study on 314 infertile women was conducted from June to December 2010. After TVS, endometrial biopsies were obtained by hysteroscopy. Pathologically confirmed EPs were taken as the gold standard.

RESULTSBased on recognized criteria, TVS had a sensitivity of 37.04%, specificity of 98.71%, positive predictive value of 90.91%, negative predictive value of 81.85%, and accuracy of 82.80% for diagnosing EPs. Mean endometrial thickness was significantly different in patients with and without EPs (P = 0.0001). In women in the mid and late-proliferative phase, the endometrial thickness was significantly greater in those with EPs than in those without them (P = 0.0001 and 0.024). Receiver operating characteristic analysis showed that endometrial thickness had a sensitivity of 85.2% and specificity of 38% in the diagnosis of EPs, the area under the curve being 0.64. In the mid-proliferative phase, sensitivity was up to 90.9%, the area under the curve being 0.70.

CONCLUSIONSTVS is poor at detecting EPs in infertile women; however, transvaginal sonographic measurement of endometrial thickness is helpful. It is suggested that the diagnostic value of TVS for EPs in infertile women could be improved by adding the measurement of endometrial thickness to the variables that are routinely assessed.

Adult ; Female ; Humans ; Infertility, Female ; diagnostic imaging ; Polyps ; diagnostic imaging ; Retrospective Studies ; Ultrasonography ; Uterine Diseases ; diagnostic imaging