The effect of amygdalin joint hydroxysafflor yellow A (HSYA) on the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and the possible mechanism were studied and explored. Chondrocytes were obtained from endplate of one-month SD rat intervertebral discs and cultured primary endplate chondrocytes. After identification, they were divided into normal group, induced group, amygdalin group, HSYA group and combined group. CCK-8 kit was adopted to detect the proliferation of the endplate chondrocytes. FCM was measured to detect the apoptosis. Real-time PCR method was adopted to observe the mRNA expression of Aggrecan, Col 2 alpha1, Col 10 alpha1, MMP-13 and the inflammatory cytokines IL-1beta. The protein expression of Col II, Col X was tested through immunofluorescence. Compared with the normal group, the proliferation of the endplate chondrocytes decreased while the apoptosis increased (P < 0.05). With down regulation of the mRNA expressions of Aggrecan, Col 2 alpha1 and up regulation of the mRNA expressions of Col 10 alpha1, MMP-13, IL-1beta (P < 0.05), the protein expression of Col II decreased while the protein expression of Col X increased. Compared with the induced group, amygdalin group, HSYA group, the combined group could inhibit the apoptosis and promote the proliferation (P < 0.05). They could increase the mRNA expressions of Aggrecan and Col 2 alpha1 while decrease the mRNA expressions of Col 10 alpha1, MMP-13 and IL-1beta (P < 0.05). They could also enhance the protein expression of Col II while reduce the protein expression of Col X. The effect of the combined group was significantly better than that of amygdalin and HSYA. Amygdalin joint HSYA could inhibit the degeneration of the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and better than the single use of amygdalin or HSYA.
Amygdalin ; pharmacology ; Animals ; Apoptosis ; Cells, Cultured ; Chalcone ; analogs & derivatives ; pharmacology ; Chondrocytes ; drug effects ; Collagen ; metabolism ; Drug Synergism ; Interleukin-1beta ; Intervertebral Disc ; cytology ; Quinones ; pharmacology ; Rats

Hepatorenal Syndrome ; diagnosis ; therapy ; Humans ; Prognosis

Previous studies demonstrated that interleukin-12 (IL-12) enhances the non-MHC-restricted cytotoxic activity of NK cells and facilitate specific allogeneic human cytotoxic T lymphocyte responses against fresh leukemia cells and cell lines. The Wilms' tumor gene, WT1 mRNA, has been used as a marker of minimal residual disease (MRD) for evaluating therapeutic efficacy of patients with leukemia or myelodysplastic syndrome (MDS). This study was aimed to investigate whether in vitro IL-12 can lower WT1 gene expression in peripheral blood monuclear cells (PBMNC) from patients with leukemia or MDS. PBMNC from these 30 patients and 5 healthy volunteers were cultured at 5 x 10(5) cells/ml alone with or without 100 units/ml of IL-12 for 3 days. WT1 mRNA was measured by competitive reverse transcription polymerase chain reaction (RT-PCR) since WT1 mRNA is considered as a marker of minimal residual disease (MRD) in leukemia and MDS. The results demonstrated that WT1 mRNA in PBMNC of 5 healthy volunteers was less than 10(3) copies/microg of total RNA. Following the 3-day IL-12 treatment, mean WT1 mRNA of PBMNC was reduced from 10(4.8) to 10(4.2) copies/microg of total RNA in 6 CML patients, from 10(5.4) to 10(4.8) copies/microg in 12 MDS patients and from 10(5.0) to 10(4.2) copies/microg in 5 AML patients in CR, but not reduced in 5 of 7 AML in non-CR. It is concluded that IL-12 significantly decrease the quantity of leukemia cells in PBMNC of most patients with MDS, CML and AML in CR. IL-12 may be of considerable benefit in the elimination of MRD in patients with hematological malignancies.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Interleukin-12 ; pharmacology ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; Neoplasm, Residual ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; WT1 Proteins ; biosynthesis ; genetics

OBJECTIVETo observe effects of removing arms and ovarian on lumbar intervertebral disc and vertebral bone mineral density (BMD) by establishing rat model of lumbar intervetebral disc degeneration (IDD) with kidney deficiency, and to explore internal mechanism of disc degeneration, relationship between disc degeneration and osteoporosis.

METHODSThirty Sprague-Dawley female rats aged one month were randomly divided into control group, lumbar IDD group and lumbar IDD with kidney deficiency group (combined group), 10 rats in each group. Lumbar IDD group removed double arms, lumbar IDD with kidney deficiency group removed double arms after 3 months, both ovaries were removed. Vertebral bone mineral density were observed by Micro-CT scan; morphological changes were tested by safranine O-fast green staining; II, X collagen protein expression in the intervertebral disc were obsevered by immunohistochemistry; extracellular matrix gene expression were obsevered by real-time polymerase chain reaction (RT-PCR), in order to evaluate the effects of removed of forelimbs and double ovarian on degeneration and vertebral bone mineral density of intervertebral disc.

RESULTSMicro-CT scan showed osteoporosis in kidney deficiency group was obviously worse than other two groups; safranine O-fast green staining showed that intervertebral space became narrowed, intervertebral disc tissue degenerated obviously, chondral palte was underdeveloped in kidney deficiency group; immunohistochemistry showed that X collagen expression increased, type II collagen expression decreased in kidney deficiency group; RT-PCR showed that type II collagen expression in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P=0.000); Age 1 in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P= 0.000); while type X collagen expression was higher than control group, but no significant meaning; MMP-13 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P= 0.000, P=0.000); aggrecanase-2 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P=0.006, P=0.008).

CONCLUSIONRats model of lumbar disc degeneration established by removed forelimbs and ovariectomized can occure "bone like"--osteoporosis, which is similar with clinical kidney lumbar disc degeneration in tissue morphology, molecular cell biology expression.

Animals ; Collagen ; genetics ; metabolism ; Extracellular Matrix ; genetics ; metabolism ; Female ; Humans ; Intervertebral Disc Degeneration ; etiology ; metabolism ; physiopathology ; surgery ; Kidney ; physiopathology ; Osteoporosis ; complications ; genetics ; metabolism ; Ovariectomy ; adverse effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the approach of open reduction and internal fixation for the treatment of anteromedial coronoid facet fractures and associated injury.

METHODSFrom July 2009 to August 2011, 6 coronoid anteromedial facet fractures were treated (4 males and 2 females,the average age was 32.6 years old,ranged from 19 to 49 years old) in our hospital. Three patients had fractures in the left side and 3 in the right(4 dominant sides and 2 non dominant sides). All the patients had close fractures. All the fractures were subtype 2 or subtype 3 of coronoid anteromedial facet fracture according to O'Driscoll classification. The anteromedial incision of elbow and the approach of splitting flexor digitorum superficialis between flexor carpi radialis and palmaris longus were used to apply internal fixation with mini-plate. After fixation of coronoid fracture, widening of radiohumeral joint interspace under varus stress occurred in 3 cases,elbow varus posteromedial rotational instability was showed,and the exploration and repair of radial collateral ligament was not carried out. Plaster was applied for 2 weeks before rehabilitation and the time of plaster application extended to 4 weeks in the patients showed varus posteromedial rotational instability. The motion degree,pain and stability, strength of elbow and hand were recorded,the elbows were evaluated with modified An and Morrey functional rating index.

RESULTSAll the patients were followed up,and the average duration was 9.3 months (ranged from 7.5 to 13 months). The strength of elbow and hand were equal to that in the contralateral side. The average flexion was (129.0+/-6.5) degree (ranged from 120 to 135 degree); the average extention was (4.0+/-4.2) degree (ranged from 0 to 10 degree); the average forearm pronation was (84.0+/-6.5) degree (ranged from 75 to 90 degree); average supination was (89.0+/-7.1) degree (ranged from 80 to 100 degree). One patient had ulnar sensory neuropathy after operation and healed in half-year, no postoperative complications as pain and instability accured. All the patients obtained excellent results according to modified An and Morrey functional rating index.

CONCLUSIONThe approach of splitting flexor digitorum superficialis between flexor carpi radialis and palmaris longus could be used in open reduction and internal fixation of anteromedial coronoid facet farcture, rehabilitation should be postponed in the patient showed elbow varus posteromedial rotational instability.

Adult ; Casts, Surgical ; Female ; Fracture Fixation, Internal ; adverse effects ; methods ; Humans ; Male ; Middle Aged ; Ulna Fractures ; surgery

OBJECTIVETo investigate the clinical effects of modified Nirschl surgical techniique in treating refractory lateral epicondylitis.

METHODSFrom March 2009 to January 2011,21 patients (21 elbows) with refractory lateral epicondylitis were treated in our hospital. There were 8 males and 13 females,ranged in age from 25 to 59 years with an average of (48.3+/-13.4) years and the duration time from 8 to 33 months with an average of (17.1+/-7.7) months;affected position in dominant sides of 16 cases and non-dominant sides of 5 cases. The patients had already received multiple non-operative treatments. Modified Nirschl surgical technique was performed,and operative origination from origin of musculus extensor carpi radialis brevis to discard process with small incision, the process place of extensor digitorum communis would be removed. The incisions were nursed by ice compress for 2 days after operation. Range of motion (ROM) and strengthening exercise of elbow joints started at the 1 week after plaster slab fixation; ROM and strengthening exercise of wrist joints also started at the 2 week after fixation. The pain, power of gripping and patient staisfaction were recorded after operation. Verhaar scaling were used to estimate the recovery.

RESULTSSeventeen patients were followed up from 13 to 22 months with an average of 16.3 months. According to Verhaar standard, 15 cases obtained excellent results and 2 good. No postoperative complication such as instability was found.

CONCLUSIONModified Nirschl surgical technique is an effective method in treating refractory lateral epicondylitis but correct to diagnosis and exclusion the coexisting diseases,accurate removal the process are important guarantee.

Adult ; Diagnosis, Differential ; Elbow Joint ; surgery ; Female ; Humans ; Male ; Middle Aged ; Tennis Elbow ; diagnosis ; etiology ; surgery

OBJECTIVETo investigate the effects of artesunate (ART) on proliferation, cell cycle and apoptosis of SKM-1 cells in vitro and to explore the underlying mechanisms.

METHODSAfter SKM-1 cells were treated with different concentrations of ART, the cell proliferation was determined by CCK-8 method. Apoptosis and distribution of cell cycle were detected by flow cytometry. Both DCFH-DA fluorescent probe and Fluo-3-Am fluorescent probe were used to detect the changes of intracellular reactive oxygen species (ROS) and calcium ion concentration. Western blot was used to measure the protein levels of BCL-2, BAX, BAD, P-BAD, survivin and XIAP.

RESULTSART obviously inhibited the growth of SKM-1 cells in time and dose-dependent manners (r = -0.841; r = 0.-786). The antioxidant trolox-pretreatment significantly decreased the growth inhibition effect of ART on SKM-1 cells. Caspase inhibitor Ac-DEVD-CHO partially reduced the growth inhibition effect of ART on SKM-1 cells. After treatment with ART for 24 hours, the apoptosis of SKM-1 cells was found, the cell cycle of SKM-1 was arrested in G0/G1 phase, ART could elevate the levels of calciumion and reactive orygen. ART could significantly down-regulate the protein expression levels of P-BAD and survivin in SKM-1 cells, and showed a highly negative correlation with ART dose (r = -0.909; r = -0.849). On the contrary, ART had no significant effect on expression levels of BAD and XIAP in SKM-1 cells, and after ART treatment, although BCL-2 protein expression was not significantly different when compared with control group, but the BCL-2/BAX ratio significantly decreased and highly negatively correlated with ART dose (r = -0.866).

CONCLUSIONThe ART significantly suppresses the cell proliferation, induces the apoptosis and promoted cell cycle arrest at G0/G1 phase in SKM-1 cells. The mechanisms of ART anti-MDS is associated with the increase of intracellular calciumion concentration and ROS levels. In addition, the pro-apoptotic activity of ART may be involved in the regulation of BCL-2 /BAX ratio and the expressions of P-bad and survivin.

Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Calcium ; metabolism ; Cell Cycle ; drug effects ; Cell Cycle Checkpoints ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Oligopeptides ; pharmacology ; Reactive Oxygen Species ; metabolism

Objective In the legal proceedings of murder cases,we often encounter the problem of proof standard in traditional forensic pathology in deep research,which is lack of systemic discussion in domestic academic field.This paper aimed to discuss this problem.Methods We compared the relevant characteristics of different legal systems on the standard of proof system,analyzed the elements of the forensic pathology evidence related to murder cases,and discussed the problem with the author's judicial practice experience.Results We believe that the cause of death,death time,injury and injury tools are the three most important forensic pathology evidences in the murder of the standard system as the "three pillars".Conclusions Division of standards in traditional forensic pathology evidence in the murder is of great significance to raise the sense of evidence of forensic medicine,and if it exists major flaws in the evidence of forensic pathology,the evidence can be corrected or the proceedings can be terminated at the very beginning of the litigation phase,which is of great value to the savings of judicial resources.

OBJECTIVETo investigate the changes and clinical value of circulating endothelial cells (CEC) in the peripheral blood of advanced NSCLC patient.

METHODSSixty-seven advanced NSCLC patients were randomly divided into either the treatment group with NP plus endostatin or control group with NP alone. Level of CEC and cytokeratin (CK) in the peripheral blood were measured by flow cytometry.

RESULTSThe response rate and benefit rate was 44.4%, 80.0% in the treatment group, and 27.3%, 50.0% in the control group, respectively (P = 0.176 and P = 0.012). Time to tumor progression (TTP) was 146.7 days in the treatment group and 91.1 days in the control group (P = 0.061). However, when the cut-off of TTP was defined as > 170 days, there was a significant difference between two groups (cut-off = 170, P = 0.034; cut-off = 180, P = 0.009). The number of CEC decreased by 0.29 +/- 0.47 in the treatment group and by 0.01 +/- 0.43 in the control group (P = 0.033). The correlation between CEC and CK was found to be positive either before (r = 0.381, P = 0.013) or after the treatment (r = 0.450, P = 0.004).

CONCLUSIONChemotherapy combined with endostatin is superior to chemotherapy alone in the treatment of NSCLC. CEC, as a biomarker, may be useful in predicting the efficacy of the combined treatment.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; blood ; drug therapy ; pathology ; Cell Count ; Cisplatin ; administration & dosage ; Endostatins ; administration & dosage ; Endothelial Cells ; pathology ; Endothelium, Vascular ; pathology ; Female ; Flow Cytometry ; Follow-Up Studies ; Humans ; Keratins ; blood ; Lung Neoplasms ; blood ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; pathology ; Remission Induction ; Treatment Outcome ; Vinblastine ; administration & dosage ; analogs & derivatives

XueBiJing is an intravenous five-herb injection used to treat sepsis in China.The study aimed to develop a liquid chromatography-tandem mass spectrometry(LC-MS/MS)-or liquid chromatography-ultraviolet(LC-UV)-based assay for quality evaluation of XueBiJing.Assay development involved identifying marker constituents to make the assay therapeutically relevant and building a reliable one-point cali-brator for monitoring the various analytes in parallel.Nine marker constituents from the five herbs were selected based on XueBiJing's chemical composition,pharmacokinetics,and pharmacodynamics.A selectivity test(for"similarity of response")was developed to identify and minimize interference by non-target constituents.Then,an intercept test was developed to fulfill"linearity through zero"for each analyte(absolute ratio of intercept to C response,＜2％).Using the newly developed assays,we analyzed samples from 33 batches of XueBiJing,manufactured over three years,and found small batch-to-batch variability in contents of the marker constituents(4.1％-14.8％),except for senkyunolide I(26.5％).