To screen and evaluate protein biomarkers for the detection of gliomas (Astrocytoma grade I-IV) from healthy individuals and gliomas from brain benign tumors by using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) coupled with an artificial neural network (ANN) algorithm. SELDI-TOF-MS protein fingerprinting of serum from 105 brain tumor patients and healthy individuals, included 28 patients with glioma (Astrocytoma I-IV), 37 patients with brain benign tumor, and 40 age-matched healthy individuals. Two thirds of the total samples of every compared pair as training set were used to set up discriminating patterns, and one third of total samples of every compared pair as test set were used to cross-validate; simultaneously, discriminate-cluster analysis derived SPSS 10.0 software was used to compare Astrocytoma grade I-II with grade III-IV ones. An accuracy of 95.7%, sensitivity of 88.9%, specificity of 100%, positive predictive value of 90% and negative predictive value of 100% were obtained in a blinded test set comparing gliomas patients with healthy individuals; an accuracy of 86.4%, sensitivity of 88.9%, specificity of 84.6%, positive predictive value of 90% and negative predictive value of 85.7% were obtained when patient's gliomas was compared with benign brain tumor. Total accuracy of 85.7%, accuracy of grade I-II Astrocytoma was 86.7%, accuracy of III-IV Astrocytoma was 84.6% were obtained when grade I-II Astrocytoma was compared with grade III-IV ones (discriminant analysis). SELDI-TOF-MS combined with bioinformatics tools, could greatly facilitate the discovery of better biomarkers. The high sensitivity and specificity achieved by the use of selected biomarkers showed great potential application for the discrimination of gliomas patients from healthy individuals and gliomas from brain benign tumors.
Adult ; Aged ; Algorithms ; Artificial Intelligence ; Astrocytoma ; blood ; classification ; diagnosis ; Biomarkers, Tumor ; blood ; Brain Neoplasms ; blood ; classification ; diagnosis ; Diagnosis, Computer-Assisted ; methods ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Proteins ; blood ; Neural Networks (Computer) ; Peptide Mapping ; methods ; Protein Array Analysis ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

OBJECTIVETo explore the efficacy and the reproductive endocrinal mechanism of herbal-partitioned moxibustion in the treatment of primary dysmenorrhea.

METHODSOne hundred and seventy-one cases of primary dysmenorrhea were randomized into an herbal-partitioned moxibustion group (group A), an starch-partitioned moxibustion group (group B) and an acupuncture group (group C), 57 cases in each one. In the group A, moxibustion isolated with herbal medicine was applied to Shenque (CV 8). In the group B, moxibustion isolated with starch was used at Shenque (CV 8). In the group C, acupuncture was given at Sanyinjiao (SP 6). The changes of estradiol (E2), progesterone (P) and prostaglandin levels (PGF2alpha) were observed before and after treatment, and the therapeutic effects were compared among the 3 groups.

RESULTSThe therapeutic effect in the group A was better than those in the other two groups [compared the cured rate: 89.8% (44/49) vs 60.0% (30/50), 60.4% (32/53), both P < 0.05]. In the group A, E2 level [(110.99 +/- 12.90) pg/mL vs (83.94 +/- 8.91) pg/mL, P < 0.05] and PGF2alpha level [(24.58 +/- 3.01) pg/mL vs (14.34 +/- 1.48) pg/mL, P < 0.01] were decreased and P level was increased [(4.65 +/- 0.68) ng/mL vs (6.68 +/- 0.95) pg/mL, P < 0.05]. In the group B and C, PGF2alpha level were reduced. Concerning to the regulating of E2 and PGF2alpha levels, the results in the group A were better than those in the group B and C [(-30.16 +/- 10.20) pg/mL vs (10.79 +/- 15.01) pg/mL, (22.81 +/- 12.22) pg/mL; (-13.10 +/- 2.40) pg/mL vs (-6.52 +/- 1.88) pg/mL, (-3.14 +/- 1.19) pg/mL, (see text) P < 0.05]. Concerning to the regulation of P level, the results in the group A and B were better than that in the group C (all P < 0.05).

CONCLUSIONThe herbal-partitioned moxibustion achieves the significant efficacy on primary dysmenorrhea, which could be related to regulating the reproductive endocrinal level. It decreases E2 and PGF2alpha levels and increases P level.

Acupuncture Points ; Adult ; Dinoprost ; metabolism ; Dysmenorrhea ; metabolism ; therapy ; Estradiol ; metabolism ; Female ; Humans ; Moxibustion ; Progesterone ; metabolism ; Young Adult

OBJECTIVETo observe the therapeutic effect of acupuncture treatment on stroke according to the acupoint selection of "Najia method of Ziwua Liuzhu" and to investigate the mechanism.

METHODSOne hundred and ninety cases were randomly divided into a Ziwu Liuzhu group (n=95) and a routine acupuncture group (n=95). Five shu points on different meridians were selected according to the Tiangan (Heavenly Stems)-tables of "Najia method of Ziwu Liuzhu" in the Ziwu Liuzhu group, and the treatment was carried out within the period from Chen (7:00-9:00 AM) to Si (9:00-11:00 AM). In the routine acupuncture group, Fengchi (GB 20), Shuigou (GV 26), Waiguan (TE 5), etc. were selected to treat the patients immediately on first visit. The therapeutic effects of two groups were assessed by the scores of neural functial cinl deficiency, status of total living ability, blood rheological indexes and clinical comprehensive effectiveness.

RESULTSThe total effective rate of 95.8% in the Ziwu Liuzhu group was significantly better than 80.0% in the routine acupuncture group. The scores of neural functional deficiency, main items of blood rheology and total living ability in the Ziwu Liuzhu group were significantly lower than those in the routine acupuncture group (all P < 0.05).

CONCLUSIONAcupuncture treatment in the period from Chen to Si according to the acupoint selection of "Najia method of Ziwu Liuzhu" has a significant clinical effectiveness which is related with improvement of the indexes of blood rheology.

Activities of Daily Living ; Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Female ; Hemorheology ; Humans ; Male ; Meridians ; Middle Aged ; Stroke ; blood ; physiopathology ; therapy

OBJECTIVETo observe the effect of simvastatin carried by poly (lactide-co-glycolide) (PLGA) on residual ridge resorption following tooth extraction.

METHODSSixty male Wistar rats were divided into experimental groups and control groups (30 rats/group). PLGA was immediately implanted with or without simvastatin into extraction sockets of the mandibular incisors. Soft X-ray photography, bone mineral density (BMD) and histopathologic study were conducted at 7, 14, 28, 56, and 84 days after implantation.

RESULTSThe relative length values of residual alveolar ridge of the experimental groups were greater than those of the controls at 14, 28, 56, and 84 days after implantation, and there was a significant difference between the experimental and control groups (P < 0.05). The BMD of the specific region was higher in the experimental groups [(7.101 +/- 0.025), (7.178 +/- 0.039), and (7.162 +/- 0.052) g/cm(2)] than that in the control groups [(7.074 +/- 0.014), (7.117 +/- 0.012), and (7.059 +/- 0.037) g/cm(2)] (P < 0.05) after 28, 56, and 84 days. Light microscopy showed that bone formation rate and quality of the experimental group were better than those in the control group at the same time.

CONCLUSIONSSimvastatin carried by PLGA could induce bone formation of tooth socket. Local application of simvastatin would be potential to preserve the length and bone volume of alveolar ridge after tooth extraction.

Alveolar Bone Loss ; prevention & control ; Alveolar Process ; pathology ; Animals ; Lactic Acid ; administration & dosage ; therapeutic use ; Male ; Polyglycolic Acid ; administration & dosage ; therapeutic use ; Rats ; Rats, Wistar ; Simvastatin ; administration & dosage ; therapeutic use ; Tooth Extraction

OBJECTIVETo investigate the effect of particulate cancellous bone impaction grafting in combination with total hip arthroplasty (THA) for acetabular reconstruction in patients with posttraumatic arthritis and bone loss after acetabular fractures.

METHODSTotally 15 consecutive cases with unilateral acetabular fracture were treated with bone impaction grafting in combination with THA in our department. There were 10 males and 5 females with mean age of 48.2 years (ranging from 36 to 73 years). Eight cases had the fracture at left hips, 7 at right hips. The average age at injury was 28 years (ranging from 18 to 68 years). The mean follow-up period was 4.3 years (ranging from 2 to 7 years).

RESULTSCompared with mean 42 points (ranging from 10 to 62) of the preoperative Harris score, the survival cases at the final follow-up had mean 84 points (ranging from 58 to 98). One patient had mild pain in the hip. No revision of the acetabular or femoral component was undertaken during the follow-up. Normal rotational centre of most hips was recovered except 2 cases in which it was 0.8 mm higher than that in opposite side. All of them had a stable radiographic appearance. Progressive radiolucent lines were observed in I, III zones in 2 cases. One patient had a nonprogressive radiolucent line in zone III. The cup prosthesis was obviously displaced (6 mm) in one patient, but had not been revised.

CONCLUSIONParticulate cancellous bone impaction grafting in combination with THA as a biological solution is an attractive procedure for acetabular reconstruction in patients with posttraumatic arthritis and bone loss after acetabular fracture, which can not only restore acetabular bone stock but also repair normal hip anatomy and its function.

Acetabulum ; surgery ; Adolescent ; Adult ; Aged ; Arthritis ; surgery ; Arthroplasty, Replacement, Hip ; Bone Substitutes ; Bone Transplantation ; methods ; Female ; Hip Fractures ; surgery ; Humans ; Male ; Middle Aged

OBJECTIVETo investigate the expression of HPA and HIF-1alpha in human esophageal squamous cell carcinoma and their relationship with cancer development, invasion and metastasis.

METHODSThe expression of HPA mRNA and HIF-1alpha protein in 23 mucosa specimens of incisal margin, 26 para-tumorous dysplastic tissues and 70 esophageal squamous cell carcinoma specimens were detected by in situ hybridization assay and immunohistochemical staining, respectively.

RESULTSThe positive expression of HPA mRNA and HIF-1alpha protein were significantly increased as the epithelial cells progressed into carcinoma (P < 0.05). The expression of HPA mRNA and HIF-1alpha protein in the esophageal squamous cell carcinoma were significantly correlated with the invasion depth, lymph node metastasis and clinical staging (P < 0.05), while it was not correlated with the differentiation of tumors (P > 0.05). There was a close correlation between the expression of HPA mRNA and HIF-1alpha protein in the carcinoma tissues (P < 0.01).

CONCLUSIONThe high expression of HPA mRNA and HIF-1alpha protein is correlated with carcinogenesis, progression, invasion and metastasis of esophageal squamous cell carcinoma. There may be a positive cooperation between two of them in the carcinogenesis and development of esophageal carcinoma. The determination of HPA mRNA and HIF-1 alpha will be useful in predicting tumor biological behavior.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Esophageal Neoplasms ; metabolism ; pathology ; Esophagus ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Glucuronidase ; genetics ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Immunohistochemistry ; In Situ Hybridization ; Lymphatic Metastasis ; Male ; Middle Aged ; Mucous Membrane ; metabolism ; Neoplasm Invasiveness ; Neoplasm Staging ; RNA, Messenger ; metabolism

This study was purposed to investigate the effects of siRNA targeting c-myc on apoptosis induction, proliferation in inhibition as well as c-myc protein and mRNA expression in human myeloid leukemia cell line HL-60 cells. C-myc siRNA synthesized in vitro was transfected into HL-60 cells by liposome. Changes of cell morphology were observed. Growth inhibition was detected by MTT assay and colony formation assay, and cell apoptosis was determined by DNA ladder. The expressions of c-myc mRNA and protein were detected by RT-PCR and Western-blot respectively. The results indicated that c-myc siRNA remarkably inhibited the cell proliferation, with an IC(50) value of 150 nmol/L. Data of DNA ladder showed that HL-60 cells apoptosis could be efficiently induced by c-myc siRNA, the apoptosis rate positively correlated with the time duration of treatment with drugs. The c-myc mRNA and protein expressions on HL-60 cells decreased after treatment with c-myc siRNA, which negatively correlated with time duration of treatment. It is concluded that c-myc siRNA can efficiently induce growth inhibition, decrease the expressions of c-myc mRNA and protein, and induce apoptosis in HL-60 cells.
Apoptosis ; genetics ; Cell Proliferation ; Gene Targeting ; HL-60 Cells ; Humans ; Leukemia, Myeloid ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo define the effects of Astragalus membranaceus on the atrial dynamics and ANP secretion in the perfused beating rabbit atria.

METHODThe experiments have been done in isolated perfused beating rabbit atria. ANP was measured by radioimmunoassay in the atrial perfusate in real-time base.

RESULTA. membranaceus (2.0, 2.5, 3.0 g L(-1)) could increase atria stroke volume from (694.70 +/- 0.01) microL g(-1) (P<0.05) to (1,003.00 +/- 8.80) microL g(-1) (P<0.001); (1,120.00 +/- 17.71) microL g(-1) and (1,195.00 +/- 8.21) microL g(-1) (P<0.001), respectively, and its could difference increase atrial pulse pressure from (0.82 +/- 0.01) kPa to (0.86 +/- 0.01) kPa (P<0.01); (0.96 +/- 0.01) kPa (P<0.001) and (1.02 +/- 0.01) kPa (P<0.001), respectively; A. membranaceus obviously increased rabbit atrial dynamics with dose-dependent manner. Simultaneously, A. membranaceus inhibited ANP secretion. Nifedipine (1.0 micromol L(-1)), a L-type Ca2+ channel inhibitor, and KB-R 7943 (10.0 micromol L(-1)), an inhibitor of reversed Na+ -Ca2+ exchanger, blocked the effects of A. membranaceus-induced augmentation of atrial dynamics but failed to modulation the inhibition of A. membranaceus on ANP secretion.

CONCLUSIONA. membranaceus increases the atrial dynamics via Na+ -Ca2+ exchanger and L-type Ca2+ channel and negatively modulates ANP secretion in beating rabbit atria.

Animals ; Astragalus membranaceus ; chemistry ; Atrial Natriuretic Factor ; metabolism ; secretion ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Heart Atria ; drug effects ; metabolism ; secretion ; Male ; Myocardial Contraction ; drug effects ; Nifedipine ; pharmacology ; Rabbits ; Radioimmunoassay ; Thiourea ; analogs & derivatives ; pharmacology

The purpose of this study was to investigate the repair of the osteoarthritis(OA)-induced cartilage injury by transfecting the new TGF-β3 fusion protein (LAP-MMP-mTGF-β3) with targeted therapy function into the bone marrow-derived mesenchymal stem cells (MSCs) in rats. The recombinant of pIRES-EGFP-MMP was constructed by combination of DNA encoding MMP enzyme cutting site and eukaryotic expression vector pIRES-EGFP. LAP and mTGF-β3 fragments were obtained from rat embryos by RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, so as to construct the recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3. pIRES-EGFP-LAP-MMP-mTGF-β3 was transfected into rat MSCs. The genetically modified MSCs were cultured in medium with MMP-1 or not. The transfected MSCs were transplanted in the rat OA models. The OA animal models were surgically induced by anterior cruciate ligament transaction (ACLT). The pathological changes were observed under a microscope by HE staining, Alcian blue, Safranin-fast Green and graded by Mankin's scale. pIRES-EGFP-LAP-MMP-mTGF-β3 was successfully constructed by means of enzyme cutting and sequencing, and the mTGF-β3 fusion protein (39 kD) was certified by Western blotting. Those genetically modified MSCs could differentiate into chondrocytes induced by MMP and secrete the relevant-matrix. The transfected MSCs could promote chondrogenesis and matrix production in rat OA models in vivo. It was concluded that a new fusion protein LAP-MMP-mTGF-β3 was constructed successfully by gene engineering, and could be used to repair the OA-induced cartilage injury.