Arthroplasty, Replacement, Knee ; methods ; Bone Transplantation ; Bone Wires ; Female ; Humans ; Middle Aged ; Tibia ; surgery ; Transplantation, Homologous

Objective To express the beta hemolysin ( Hlb), an important toxin secreted by Staphylococcus aureus ( S.aureus) and the mutant protein Hlb H-149-N , to detect the hemolytic activity of Hlb and Hlb H-149-N on sheep erythrocytes , and to prepare the specific antibodies against Hlb which can inhibit the hemolytic activity of Hlb .Methods Hlb gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as template.The expression vector pET-28a-hlb was constructed and transformed into E.coli BL21(DE3).The expression vector pET28a-hlbH-149-Nwas constructed through point mutation.The recombinant Hlb and Hlb H-149-N protein were expressed and purified by Ni 2+affinity chromatography .The hemolytic activity of Hlb and Hlb H-149-N was measured by sheep erythrocyte lysis assay .Results Recombinant Hlb protein and the mutant were obtained .Further investigations showed that Hlb could significantly induce the lysis of SRBC while HlbH-149-N could not.The specific polyclonal antibodies against Hlb (anti-Hlb) were prepared.It was found that anti-Hlb recognized Hlb and Hlb H-149-N .Moreover , it was found that anti-Hlb blocked the hemolytic activity of Hlb .Conclusion The recombinant Hlb protein with high hemolytic activity and Hlb H-149-N without hemolytic activity are obtained while its neutralized antibody is pepared .Hlb from S.aureus has different hemolytic effects on erythrocytes from various species .Our findings will facilitate the investigation on the role of Hlb in the pathogenesis of S.aureus.

Objective To clone and express alpha hemolysin ( Hla ) , one important virulence factor secreted by Staphylococcus aureus in Escherichia coli to tdetect the hemolytic activity of Hla on erythrocytes from diffrerent species ,and to prepare specific antibodies against Hla that can inhibit the hemolytic activity of Hla .Methods Hla gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as a template.The expression vector pET-28a-Hla was constructed and transformed into E.coli BL21(DE3).The recombinant Hla protein was expressed by IPTG induction ,and then purified by Ni2+affinity chromatography .The hemolytic activity of Hla was measured by erythrocyte lysis assays .Results Soluble recombinant Hla protein was expressed and purified .Further investigations showed that Hla could significantly induce the lysis of rabbit erythrocytes .However , erythrocytes from humans or sheep were more resistant to the lysis mediated by Hla . The specific polyclonal antibodies against Hla ( anti-Hla) were prepared and detected by ELISA .Moreover, it was found that anti-Hla could inhibit the hemolytic activity of Hla .Conclusion We found that Hla from S.aureus has different hemolytic effects on erythrocytes from various species .The prepared Hla-antibodies can specifically block the hemolytic activity of Hla.

The prescriptions including Polygoni Multiflori Caulis that built by Pro. Yan were collected to build a database based on traditional Chinese medicine (TCM) inheritance assist system. The method of association rules with apriori algorithm was used to achieve frequency of single medicine, frequency of drug combinations, association rules between drugs and core drug combinations. The datamining results indicated that in the prescriptions that including Polygoni Multiflori Caulis, the highest frequency used drugs were parched Ziziphi Spinosae Semen, Ostreae Concha, Ossis Mastodi Fossilia, Salviae Miltiorrhizae Radix Et Rhizoma, Paeoniae Rubra Radix, and so on. The most frequent drug combinations were "Polygoni Multiflori Caulis-parched Ziziphi Spinosae Semen", "Ostreae Concha-Polygoni Multiflori Caulis", and "Polygoni Multiflori Caulis-Ossis Mastodi Fossilia". The drug association rules of confidence coefficient 1 were "Ostreae Concha-->Polygoni Multiflori Caulis", "Poria-->Polygoni Multiflori Caulis", "parched Ziziphi Spinosae Semen-->Polygoni Multiflori Caulis", and "Paeoniae Alba Radix-->Polygoni Multiflori Caulis". The core drug combinations in the treatment of insomnia were Ossis Mastodi Fossilia, Polygoni Multiflori Caulis, Salviae Miltiorrhizae Radix et Rhizoma, Ostreae Concha, Polygalae Radix, Margaritifera Concha, Poria, and parched Ziziphi Spinosae Semen. And the core drug combinations in the treatment of obstruction of Qi in chest were Salviae Miltiorrhizae Radix Et Rhizoma, Polygoni Multiflori Caulis, parched Ziziphi Spinosae Semen, Trichosanthis Fructus, Allii Macrostemonis Bulbus, and Paeoniae Rubra Radix.
Data Mining ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Medicine, Chinese Traditional ; methods ; Plant Stems ; chemistry ; Polygonaceae ; chemistry ; Prescriptions

To study the chemical constituents of the inflorescences of Coreopsis tinctoria from Xinjiang, isolation and purification of constituents were carried out by column chromatography on macroporous resin (D101) , MCI gel, MDS gel, silica gel, Sephadex LH-20 and semi-preparative HPLC. The structures of the compounds were identified by physicchemical properties and spectral data analysis. Fourteen compounds were isolated and identified as coretinterpenoid A (1), coretinphenol (2), quercetin (3), quercetin-3-O-β-glucopyranoside (4), luteolin (5), taxifolin (6), 7, 3', 5'-trihydroxyflavanone (7), isookanin (8), isookanin-7-O-β-D-glucopyranoside (9), 5, 7, 3', 5'-tetrahydroxyflavanone-7-O-β-D-glucopyranoside (10), butein (11), okanin (12), sulfuretin (13), and linocinnamarin (14). Compound 1 was a new isabolane-type sesquiterpenoid and compounds 4, 10 and 13 were isolated from this plant for the first time.
Coreopsis ; chemistry ; Sesquiterpenes ; chemistry ; isolation & purification

BACKGROUNDTissue inhibitor of matrix metalloproteinase-1 (TIMP-1) is related to the aging of many organs, but few data are available on the change of TIMP-1 in liver aging. The purpose of this study was to investigate the expression and role of TIMP-1, matrix metalloproteinase-2 (MMP-2) and MMP-9 in the process of natural aging in the livers of normal and transgenic mice, and to detect the effects of TIMP-1 on oxidative level and anti-oxidative ability of the livers of transgenic young mice.

METHODSNormal and transgenic mice were divided into 3 groups according to their age: 3-month-old group (n = 5), 12-month-old group (n = 5) and 24-month-old group (n = 5). Histopathological changes of the liver were observed after HE and Masson staining. The messenger RNA (mRNA) levels of TIMP-1, MMP-2 and MMP-9 were determined by semi-quantitative reverse transcriptional polymerase chain reaction; protein expression was measured by Western blot in the livers of normal and transgenic mice of various ages. Changes in levels of superoxide dismutase (SOD), monoamine oxidase (MAO), malondialdehyde (MDA) as well as oxidative and anti-oxidative ability were measured.

RESULTSHistologically, more fatty degeneration and collagen deposition were found in the aging livers of transgenic mice than in those of the normal mice as their age of months increased. The mRNA and protein expressions of TIMP-1 were significantly high in the oldest animals. The histopathological changes, mRNA and protein expressions of TIMP-1 increased significantly in the liver of transgenic mice as compared with normal mice. The expression of MMP-2 and MMP-9 showed a minor change in the process of aging. Liver change and collagen deposition were not observed in young mice, but the activity of SOD decreased (P < 0.05), and the activity of MAO (P < 0.01) and the content of MDA increased in the liver of transgenic mice (P < 0.01).

CONCLUSIONSThe expression of TIMP-1 is significantly high in the liver of transgenic mouse in the process of aging, indicating that the oxidative level increases and the anti-oxidative ability decreases in the liver of transgenic mouse. TIMP-1 plays an important role in the process of liver aging.

Aging ; metabolism ; Animals ; Female ; Liver ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; analysis ; genetics ; Matrix Metalloproteinase 9 ; analysis ; genetics ; Mice ; Mice, Transgenic ; Monoamine Oxidase ; analysis ; RNA, Messenger ; analysis ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; analysis ; genetics

OBJECTIVETo evaluate the therapeutic efficacy of combined treatment with surgical decompression and drug for traumatic optic neuropathy (TON) and analyze the influential factors.

METHODSA retrospective study on 69 patients (70 eyes) with TON treated with optic canal decompression through transnasal endoscopic approach and drug was conducted. The visual acuity was divided into 5 grades: no light perception (NLP), light perception (LP), hand move, count finger, > 0.02, marked as I-V respectively. Of 40 eyes with grade I, 18 eyes received emergency operation for severe optic canal fracture confirmed by CT; 22 eyes received corticosteroid therapy firstly and then operation. Of 30 eyes above grade I, 16 eyes with optic canal fracture confirmed by CT received emergency operation; 14 eyes received corticosteroid therapy firstly and 3 days later received operation. Postoperative follow-up lasted 3-12 months to observe the recovery of visual acuity.

RESULTSThe therapeutic efficacy of patients with the visual acuity of LP and above LP was better than that of NLP (90.0% to 27.5%), the difference had statistical significance (chi(2) = 26.98, P < 0.001). In operated group, the therapeutic efficacy in patients whose visual acuity was improved from NLP after glucocorticoid therapy (80.0%) was better than that of the patients with no improvement (5.9%), the difference had statistical significance (chi(2) = 12.09, P < 0.001).

CONCLUSIONThe imaging findings of optic canal fracture can not be used as determinants for operation. The patients with NLP whose visual acuity had no improvement after corticosteroid therapy are poor candidates for surgical decompression. The visual acuity before treatment is the main factor affecting the therapeutic efficacy.

Adolescent ; Adult ; Child ; Combined Modality Therapy ; Decompression, Surgical ; Female ; Humans ; Male ; Middle Aged ; Optic Nerve Injuries ; drug therapy ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the effect of small interfering RNA (siRNA) targeting human vascular endothelial growth factor (hVEGF) on A549 cell growth in nude mice and angiogenesis on chorioallantoic membrane (CAM) assay.

METHODSThree pairs of hVEGF siRNA-plasmid and non-silencing-plasmid were constructed, and transfected into A549 cells through lipofectamine 2000, respectively. The most effective pair of hVEGF siRNA-plasmid was selected by ELISA and real-time RT-PCR. A549 cells transfected with selected hVEGF siRNA- plasmid, A549 cells transfected with non-silencing-plasmid and A549 cells without transfection were inoculated into nude mice, respectively. Chick embryos were randomly divided into four groups and CAM was treated by different solutions for 48 h: culture media DMEM as negative control group,un-transfected A549 cell culture supernatants as positive control group, hVEGF siRNA A549 cell culture supernatants as hVEGF siRNA group and nonsilencing siRNA A549 cell culture supernatants as non-silencing siRNA group. The CAMs were harvested on d12 for microscopic assays.

RESULTCompared with control group, hVEGF siRNA-plasmid induced 48% reduction in hVEGF secretion by A549 cells accompanied by 70% reduction in hVEGF mRNA. Compared with non-silencing siRNA group, the mean tumor volume of murine xenograft was reduced by 58% in hVEGF siRNA group; time for xenografts growing to 50 mm(3)was delayed by 5.4 d. hVEGF contents in xenograft were reduced by 54%; but mean doubling time of tumors and the growth rate of tumors were not significantly reduced. In CAM assays, hVEGF content was zero in negative group, and in hVEGF siRNA group that was 40%-44% of non-silencing siRNA group or positive group; vessels branch points of CAM in hVEGF siRNA group or non-silencing siRNA group or positive group were increased by 45%-55% compared with negative group; total vessel length of CAM in hVEGF siRNA group was increased by 53% compared with negative group, while in non-silencing siRNA group or positive group that was increased by 97% or 99%. Compared with negative control group, the proliferation of microvessels was increased when cell culture supernatant with hVEGF was added in hVEGF siRNA group, significant proliferated vessels were observed in non-silencing siRNA group or positive group.

CONCLUSIONA plasmid-mediated hVEGF siRNA has been constructed and verified, which can effectively downregulate the expression of hVEGF in human A549 cells, resulting in the inhibition of angiogenesis. hVEGF siRNA can delay initial growth of A549 tumor xenograft but not reduce the growth rate.

Adenocarcinoma ; pathology ; Animals ; Cell Line, Tumor ; Cell Proliferation ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; metabolism ; Female ; Humans ; Lung Neoplasms ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Physiologic ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Vascular Endothelial Growth Factor A ; antagonists & inhibitors ; genetics

OBJECTIVETo assess the clinical effect and safety of the Chinese medicine Longbishu Capsule combined with mesylate doxazosin in the treatment of benign prostatic hyperplasia (BPH) of the kidney deficiency and blood stagnation type.

METHODSThis was a randomized, double-blind, double-simulation control study. We equally assigned 60 men diagnosed with BPH of the kidney deficiency and blood stagnation type to an experimental and a control group, the former treated with mesylate doxazosin plus Longbishu Capsule and the latter with mesylate doxazosin plus placebo. We compared the International Prostate Symptom Score (IPSS), quality of life (QOL), Chinese symptom score (CSS), maximal urinary flow rate (Qmax), and prostate volume between the two groups of patients before and after 6 months of medication.

RESULTSAfter treatment, there were 5 cured cases, 13 markedly effective cases, 9 effective cases, 1 ineffective case, and 2 eliminated cases in the experimental group, as compared with 2 cured cases, 8 markedly effective cases, 10 effective cases, 7 ineffective cases, and 3 eliminated cases in the control group. The total effectiveness rate was obviously higher in the former (96.4%) than in the latter (74.1%). IPSS, Qmax, and CSS were improved in both of the groups after medication, even more significantly in the experimental than in the control group (IPSS: 15.22 ± 2.98 vs 18.15 ± 5.88, P <0.05; Qmax: [13.56 ± 2.26] ml/s vs [11.78 ± 2.97] ml/s, P <0.05; CSS: 6.18 ± 2.13 vs 9.52 ± 3.15, P <0.05). Because of the difference in the QOL score between the two groups at the baseline (P = 0.038 <0.05), no more comparison was made in this aspect after treatment.

CONCLUSIONThe combination of Longbishu Capsule with mesylate doxazosin is safe and effective for the treatment of BPH.

Adrenergic alpha-Antagonists ; therapeutic use ; Capsules ; Double-Blind Method ; Doxazosin ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Prostatic Hyperplasia ; drug therapy ; physiopathology ; Quality of Life ; Treatment Outcome ; Urination

OBJECTIVETo explore the characteristics of closed abdominal injury in pregnancy women and its treatment.

METHODSThe clinical data of 37 pregnancy patients with closed abdominal injury treated in our hospital from June 1993 to June 2003 were collected and analyzed.

RESULTSAll the 37 patients were treated with operation. Among them 2 early pregnancy patients with intestinal rupture and 1 patient with retroperitoneal hematoma were treated under laparoscope; in other 34 pregnancy patients laparotomy was performed. Of the 34 patients 8 used cesarean section because premature separation of placenta and enlarged womb interrupted the management of intra-abdominal organ injury. In the 37 patients 33 (89.1%) were cured, 4 (10.8%) die, postoperative complication rate was 16.2% (6/37). Two patients (5.4%) suffered from abdominal cavity infection, 3 (8.1%) from pulmonary infection, and 1 (2.7%) had multi-organ failure.

CONCLUSIONSFor pregnancy patients with closed abdominal injury, besides obsteric diseases intra-abdominal injury should be given much attention. Accurate diagnosis and timely treatment can gain the time to save the life of both mother and fetus.

Abdominal Injuries ; diagnosis ; surgery ; Adult ; Female ; Humans ; Laparoscopy ; Pregnancy ; Pregnancy Complications ; diagnosis ; surgery