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OBJECTIVE: To review the effectiveness of different physical therapies for acute and sub-acute low back pain supported by evidence, and create clinical recommendations and expert consensus for physiotherapists on clinical prescriptions. DATA SOURCES: A systematic search was conducted in PubMed and the Cochrane Library for studies published within the previous 15 years. REVIEW METHODS: Systematic review and meta-analysis, randomized controlled trials assessing patients with acute and sub-acute low back pain were included. Two reviewers independently screened relevant studies using the same inclusion criteria. The Physiotherapy Evidence Database and the Assessment of Multiple Systematic Reviews tool were used to grade the quality assessment of randomized controlled trials and systematic reviews, respectively. The final recommendation grades were based on the consensus discussion results of the Delphi of 22 international experts. RESULTS: Twenty-one systematic reviews and 21 randomized controlled trials were included. Spinal manipulative therapy and low-level laser therapy are recommended for acute low back pain. Core stability exercise/motor control, spinal manipulative therapy, and massage can be used to treat sub-acute low back pain. CONCLUSIONS: The consensus statements provided medical staff with appliable recommendations of physical therapy for acute and sub-acute low back pain. This consensus statement will require regular updates after 5-10 years.
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Dor Lombar , Modalidades de Fisioterapia , Humanos , Dor Lombar/reabilitação , Dor Lombar/terapia , Consenso , Ensaios Clínicos Controlados Aleatórios como Assunto , Feminino , Dor Aguda/terapia , Dor Aguda/reabilitação , MasculinoRESUMO
Objective: To systematically evaluate the clinical effectiveness of conservative treatments including pharmacological treatments and nonpharmacological treatments on patients with lumbar spinal stenosis. Methods: We searched six electronic databases systematically for randomized clinical trials published between January 2000 and July 2021, including the China National Knowledge Infrastructure, WanFang Data, PubMed, MEDLINE, Embase, and the Cochrane library. The studies focused on the therapeutic effects of pharmacological treatments including calcitonin, antiepileptics, neurotrophic drugs, nonsteroidal anti-inflammatory drugs, Chinese Traditional Medicine, limaprost, and nonpharmacological treatments like physiotherapy for treating lumbar spinal stenosis were included. The outcome was measured using the visual analog scale, Oswestry Dysfunction Index, Japanese Orthopaedic Association Score, and EuroQol Five Dimensions Questionnaire. The quality of eligible studies was assessed by using the Cochrane recommended bias risk assessment tool. Stata was used to conduct the network meta-analysis. Results: A total of 12 randomized control trials with 1,194 patients were included. The network meta-analysis showed that for the visual analog scale, a better therapeutic effect was noted while using Chinese Traditional Medicine and physiotherapy, followed by analgesics drugs and limaprost. Limaprost and calcitonin were better in decreasing the Oswestry Dysfunction Index. In terms of the Japanese Orthopaedic Association Score, the use of traditional Chinese Medicine and limaprost were associated with a better improvement than other treatments. Meanwhile, limaprost combined with analgesics drugs was found to be effective to improve the EuroQol Five Dimensions Questionnaire. Conclusion: Among the commonly used conservative treatments for the treatment of lumbar spinal stenosis, limaprost may have better efficacy in improving the Japanese Orthopaedic Association Score and decreasing the Oswestry Dysfunction Index, with a beneficial effect on decreasing the visual analog scale and improving the EuroQol Five Dimensions Questionnaire. Systematic Review Registration: website, identifier registration number.
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Objective To observe the effects of Guilu Erxian Glue (GEG) containing serum on osteogenin differentiation of bone marrow mesenchymal stem cells (BMMSCs) and Wnt signal pathway related factors. Methods Totally 100 three months old female SD rats had their bilateral ovaries excised peritoneally They were divided into the low, middle, high GEG groups, and the blank control group by random digit table, 25 in each group. The dose of GEG was calculated according to body surface area, and GEG containing serum was administered by gastrogavage for 7 successive days. Blood was collect- ed by abdominal aorta to prepare drug containing serum. F3 passage BMMSCs of 1-month SD rats were i- solated by whole bone marrow adherent method, and cultured in vitro for 3 passages. The cell surface markers (CD45 and CD90) of F3 passage were detected by flow cytometry (FCM). BMMSCs were trea- ted with different concentrations GEG containing serum for 72 h. Then the cell cycle was determined by FCM, and the proliferation index calculated. The optimal intervention concentration was determined. Then F3 passage BMMSCs were divided into four groups, i.e., the fetal bovine serum (FBS) group, the blank control group, the GEG group, the classical induction group. After they were treated with corresponding medium for 21 days, BMMSCs were dyed with alizarin red staining (ARS) to observe their osteogenin dif- ferentiation. mRNA expressions of alkaline phosphatase (ALP) and osteocalcin (OC) of BMMSCs were detected by RT-PCR. mRNA and protein expressions of Wnt5a, ß-catenin, and lymphoid enhancer factor- 1 (Lef-1) were detected by RT-PCR and Western blot. Results The ratio of CD45 positive expression was 1. 46% ?0. 23%, and the ratio of CD90 positive expression was 96. 97% ±3. 21%. Middle EGE contai- ning serum (10%) could significantly stimulate the proliferation of BMMSCs. In ARS citrus red calcium nodules could be seen in the GEG group and the classical induction group. Compared with the FBS group and the blank control group, mRNA expressions of OC and ALP were up-regulated, mRNA and protein expressions of Wnt5a and p-catenin were up-regulated in the GEG group and the classical induction group (P<0. 05). Compared with the FBS group, the blank control group, and the classical induction group, mRNA and protein expressions of Lef-1 were up-regulated in the EGE group (P <0. 05). Compared with the FBS group and the blank control group, protein expressions of Lef-1 increased in the classical induc- tion group (P <0. 05). Conclusions GEG containing serum had the functions of stimulating the prolifera- tion of BMMSCs, and inducing the osteogenic differentiation of BMMSCs. Its mechanism might be possi- bly related with regulating Wnt signal pathway related factors.
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Células-Tronco Mesenquimais , Osteogênese , Via de Sinalização Wnt , Adesivos/farmacologia , Animais , Diferenciação Celular , Células Cultivadas , Feminino , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Via de Sinalização Wnt/efeitos dos fármacosRESUMO
Brachial plexus injury (BPI) often involves the complete or partial avulsion of one or more of the cervical nerve roots, which leads to permanent paralysis of the innervated muscles. Reimplantation surgery has been attempted as a clinical treatment for brachial plexus root avulsion but has failed to achieve complete functional recovery. Lithium is a mood stabilizer drug that is used to treat bipolar disorder; however, its effects on spinal cord or peripheral nerve injuries have also been reported. The purpose of this study was to investigate whether lithium can improve functional motor recovery after ventral root avulsion and reimplantation in a rat model of BPI. The results showed that systemic treatment with a clinical dose of lithium promoted motor neuron outgrowth and increased the efficiency of motor unit regeneration through enhanced remyelination. An analysis of myelin-associated genes showed that the effects of lithium started during the early phase of remyelination and persisted through the late stage of the process. Efficient remyelination of the regenerated axons in the lithium-treated rats led to an earlier functional recovery. Therefore, we demonstrated that lithium might be a potential clinical treatment for BPI in combination with reimplantation surgery.
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Axônios/efeitos dos fármacos , Compostos de Lítio/farmacologia , Bainha de Mielina/efeitos dos fármacos , Regeneração Nervosa/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Raízes Nervosas Espinhais/efeitos dos fármacos , Animais , Axônios/patologia , Axônios/fisiologia , Plexo Braquial/efeitos dos fármacos , Plexo Braquial/lesões , Plexo Braquial/fisiopatologia , Plexo Braquial/cirurgia , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Feminino , Neurônios Motores/efeitos dos fármacos , Neurônios Motores/patologia , Neurônios Motores/fisiologia , Bainha de Mielina/patologia , Bainha de Mielina/fisiologia , Regeneração Nervosa/fisiologia , Distribuição Aleatória , Ratos Sprague-Dawley , Recuperação de Função Fisiológica/efeitos dos fármacos , Recuperação de Função Fisiológica/fisiologia , Reimplante , Raízes Nervosas Espinhais/lesões , Raízes Nervosas Espinhais/fisiopatologia , Raízes Nervosas Espinhais/cirurgiaRESUMO
BACKGROUND: Ethylene diamine tetraacetic acid dependent pseudothrombocytopenia (EDTA-PTCP) is a laboratory artifact that may lead to unnecessary evaluation and treatment of patients. The purpose of this article is to discuss how to identify EDTA-PTCP and correct spurious low platelet counts in clinical laboratories. METHODS: We use two criteria to screen for platelet aggregation: (1) an abnormal platelet count in EDTA-treated blood from a patient lacking clinical signs of a platelet disorder, and (2) an instrument flag for platelet clumps. EDTA-PTCP was confirmed by microscopic examination for platelet agglutination and by platelet counts that corrected with citrate sample. In addition, the time course of EDTA-PTCP was investigated in samples from 26 patients anticoagulated with EDTA-K2 and sodium citrate. Amikacin (5 mg/ml) was added to tubes with EDTA-K2 or sodium citrate from seven additional cases in order to confirm its dissociative effect on platelet aggregation. RESULTS: In our laboratory, the overall incidence of EDTA-PTCP was approximately 0.09%; and the duration was between 2 weeks and 6 months. EDTA-PTCP was time-dependent and occurred as early as 10 min after sample collection. Weaker agglutination could also occur in most corresponding citrate-treated samples. The dissociative effect of amikacin on platelet agglutination was case-specific and not concentration-dependent. CONCLUSIONS: The method of screening for platelet clumping with the help of XE5000 images is convenient. The decline in the platelet count is related to the length of time and the intensity of chelation. Amikacin supplement is not always effective for correcting platelet counts in vitro.
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Artefatos , Ácido Edético/química , Contagem de Plaquetas , Trombocitopenia , Erros de Diagnóstico , Humanos , Microscopia , Agregação Plaquetária , Contagem de Plaquetas/métodos , Contagem de Plaquetas/normas , Contagem de Plaquetas/estatística & dados numéricosRESUMO
The mTOR pathway is a central control of cell growth, proliferation, metabolism, and survival, and is deregulated in most cancers. Cancer cells are addicted to increased activity of mTOR kinase-mediated signaling pathways, leading to numerous inhibitors of mTOR signaling in preclinic and clinical trials for cancer therapy. Phosphorus-containing sirolimus (FIM-A), which targets mTOR signaling, inhibits cancer cell growth in vitro. Here we report that FIM-A reduces the angiogenesis and proliferation of osteosarcoma both in vitro and in vivo. In cultured osteosarcoma cell lines, FIM-A inhibited cell proliferation and arrested cells in the G1 phase of the cell cycle, accompanied with reduction of VEGF and HIF-1alpha. With in vivo mouse osteosarcoma xenografts, FIM-A treatment resulted in the inhibition of mTORC1 signaling as demonstrated by the decreased phosphorylation of p70S6K1 and 4E-BP1. Consistent with this finding, FIM-A significantly decreased the average tumor volume, nuclei staining of PCNA, and the number of intratumoral microvessels. Our data demonstrated that targeting mTORC1 by FIM-A inhibited the growth of osteosarcoma in vitro and in vivo, providing the basis for further development of FIM-A as a therapy for osteosarcoma patients.
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Antineoplásicos/farmacologia , Neoplasias Ósseas/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Neovascularização Patológica/tratamento farmacológico , Osteossarcoma/tratamento farmacológico , Sirolimo/análogos & derivados , Inibidores da Angiogênese/farmacologia , Animais , Linhagem Celular Tumoral , Feminino , Humanos , Alvo Mecanístico do Complexo 1 de Rapamicina , Camundongos , Camundongos Endogâmicos BALB C , Complexos Multiproteicos/antagonistas & inibidores , Fósforo , Sirolimo/química , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/antagonistas & inibidores , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
In this study, a high performance liquid chromatography-photo diode array detection-electrospray ionization tandem mass spectrometry (HPLC-DAD/ESI-MS) with pre-column DPPH assay is developed for screening the antioxidant components in the fruits of Livistona chinensis R. Br. Accordingly, six antioxidative flavonoids are identified as orientin, isoorientin, vitexin, isovitexin, isorhamnetin-3-O-glucoside and tricin in methanolic extract of L. chinensis fruits, based on their mass spectra and fragmentation patterns. To the best of our knowledge, orientin, isoorientin, isovitexin and isorhamnetin-3-O-glucoside were found firstly in this plant. The free radical scavenging activity of the six antioxidants found is further examined by off-line DPPH assay. The results indicated that the free radical scavenging activity of orientin and isoorientin are stronger than those of two antioxidative drugs, vitamin C and baicalin. In addition, an HPLC-DAD method is firstly established for simultaneous determination of the six antioxidants in L. chinensis fruits. Tricin was found to be the major component in L. chinensis fruits.
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Antioxidantes/análise , Arecaceae/química , Cromatografia Líquida de Alta Pressão/métodos , Flavonoides/análise , Extratos Vegetais/análise , Espectrometria de Massas por Ionização por Electrospray/métodos , Compostos de Bifenilo/análise , Frutas/química , Picratos/análise , Espectrometria de Massas por Ionização por Electrospray/instrumentaçãoRESUMO
Scutellaria barbata D. Don is widely used as a folk antitumor and anti-inflammatory agent in Asia. However, a simple and global quality control method for S. barbata was lacking. In this study, six phenolic compounds, including P-coumaric acid, scutellarin, apigenin 5-O-ß-glucopyranoside, luteolin, apigenin and 4'-hydroxywogonin were obtained from S. barbata by phytochemical investigations. The six compounds plus baicalein show cytotoxcities to the nine human cancer cells, K562, MGC-803, HL60, SH-SY5Y, SW1116, SMMC-7221, SW480, HepG2 and KB. Subsequently, a high-performance liquid chromatography with photodiode array detector (HPLC-DAD) was developed for both fingerprint analysis of S. barbata and quantitative determination of the seven anticancer active compounds in S. barbata. The chromatographic separation was accomplished on an Ultimate™ XB-C18 column (4.6 x 250 mm, 5 µm) in 65 min. For fingerprinting, 26 common peaks were found and selected as characteristic peaks to assess the consistency of S. barbata samples. For quantitative analysis, the seven bioactive compounds showed good regression relationship ( R² > 0.999) within test ranges and the recovery of the method was in the range of 90-105 %. In brief, the present study provides the fingerprint analysis and quantitative methods for global and systematical quality control of S. barbata for its anticancer usage.
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Antineoplásicos Fitogênicos/análise , Flavonoides/análise , Neoplasias/tratamento farmacológico , Fenóis/análise , Fitoterapia , Extratos Vegetais/química , Controle de Qualidade , Scutellaria/química , Antineoplásicos Fitogênicos/farmacologia , Antineoplásicos Fitogênicos/uso terapêutico , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão/métodos , Flavonoides/farmacologia , Flavonoides/uso terapêutico , Humanos , Fenóis/farmacologia , Fenóis/uso terapêutico , PolifenóisRESUMO
BACKGROUND: This is a prospective clinical study based on a large sample gathered from multiple centers in China, subordinating to 10th Five-Year Plan of National Science & Technology Progression. We analyzed the high-risk factors inducing hypertensive disorders in pregnancy (HDP) and estimated the potential effect of anti-oxidants administration, including vitamin C (VC), vitamin E (VE) and Salvia Miltiorrhiza L (SML), a Chinese herb medicine, in amelioration of the high-risk factors in pregnancy. METHODS: From April 2005 to July 2006, 4814 pregnant women from 24 national wide cooperative hospitals were involved in this prospective research. The participants were randomly divided into two groups: 1607 cases were in anti-oxidants group with administration of vitamins and SML; 3207 cases were in control group without any medicine given. Every participant was under monitoring for the morbidity of HDP and the high-risk factors were investigated in HDP cases in each group. RESULTS: (1) The morbidity of HDP was 3.55% in anti-oxidants group vs. 4.18% in control group. No statistical difference existed between the two groups (P > 0.05). (2) In anti-oxidants group, the HDP morbidities among three subgroups: VC + VE + SML, VC + VE and SML only, were 5.51%, 3.05% and 5% respectively. It showed no statistical difference among three remedies (P > 0.05). (3) The related index of factors affecting HDP showed in intensity sequence as follows: family HDP history > profession > education level > age > body weight. The incidence of HDP in normal population was 3.51%, and the incidence of HDP in high-risk pregnant women (family HDP history, heavy physical labor, low education level (middle school and below), age ≥ 40, body mass index ≥ 24) was 5.84%, which was obviously higher than that in normal population (P < 0.01). In anti-oxidants group, the probability of HDP in women with high-risk factors was 3.81%, which was obviously lower than that in control group with high-risk factors at 7.14% (P < 0.01). (4) In control group, the morbidity of HDP in women with family HDP history (especially with sisters'), heavy physical labor, middle school and below, age ≥ 35 was: 50.00%, 15.22%, 6.33%, 26.28% and 5.75%, respectively, and that in anti-oxidants group was 0, 7.69%, 3.74%, 9.27% and 2.67%, respectively, which was obviously lower than that in control group. CONCLUSIONS: The high-risk factors prone to induce HDP included: family history of HDP, heavy physical labor, low education level, aging and obesity. No impressive effect of anti-oxidants application was found in preventing HDP in general population but the remedy demonstrated positive effect on preventing HDP in pregnant women with high-risk factors.
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Antioxidantes/uso terapêutico , Hipertensão/tratamento farmacológico , Complicações na Gravidez/tratamento farmacológico , Adolescente , Adulto , Fatores Etários , Peso Corporal , Feminino , Humanos , Hipertensão/fisiopatologia , Modelos Logísticos , Gravidez , Estudos Prospectivos , Fatores de Risco , Adulto JovemRESUMO
BACKGROUND: Tai chi (TC) is proposed as a potential option for the management of osteoarthritis (OA), however, its beneficial effect on patients with knee OA has not been convincing. OBJECTIVES: To evaluate the effect of a 24-week TC program on physical functions in older Chinese women with knee OA. METHODS: Thirty-five older Chinese women with knee OA were randomized into TC group (n = 18) and attention control (wellness education and stretching) group (n = 17). Subjects in the TC group practiced the 24-form simplified Yang-style TC 2 to 4 times a week for 24 weeks with frequency gradually increased. Physical function was assessed using the Western Ontario and McMaster University Osteoarthritis Index (WOMAC), 6-minute walk distance and stair climb time. RESULTS: Compared with the control group, the participants in TC group had statistically significant improvements in changes of the WOMAC total score (6.18 +/- 2.13 vs. 1.71 +/- 2.73, P = 0.000), the WOMAC pain subscale (1.36 +/- 0.22 vs. 0.07 +/- 1.00, P = 0.001), the WOMAC stiffness subscale (0.66 +/- 0.25 vs. 0.05 +/- 0.38, P = 0.043), the WOMAC function subscale (6.17 +/- 1.96 vs. 1.72 +/- 2.63, P = 0.000), the 6-minute walk distance (32.43 +/- 14.20 vs. 6.67 +/- 16.76, P = 0.003), and the stair climb time (2.27 +/- 0.74 vs. 0.27 +/- 1.24, P = 0.001). CONCLUSIONS: This study suggests that TC provides a safe, feasible and useful exercise option for older Chinese female patients with knee OA.
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Osteoartrite do Joelho/reabilitação , Tai Chi Chuan , Idoso , China , Feminino , Humanos , Pessoa de Meia-Idade , Osteoartrite do Joelho/fisiopatologia , Recuperação de Função FisiológicaRESUMO
OBJECTIVE: To evaluate the efficacy and safety of motherwort (herba leonuri/leonurus heterophyllus sweet) injection for preventing postpartum hemorrhage after caesarian section. METHODS: The prospective study was designed as a randomized and single blind multi-center research matched with positive agent as controls from Apr 2007 to Aug 2007. 440 women underwent caesarian section (CS) indicated by obstetric factors were enrolled from 15 teaching hospitals in China and assigned into three groups: group of motherwort: 147 cases were administered by motherwort 40 mg uterine injection during CS and 20 mg intramuscular injection per 12 hours 3 times after CS; group of motherwort+oxytocin: 144 cases were administered by motherwort 40 mg and oxytocin 10 U uterine injection during CS and motherwort 20 mg intramuscular injection per 12 hours 3 times after CS and group of oxytocin: 149 cases were administered by oxytocin 10 U uterine injection and oxytocin 10 U+5% glucose 500 ml intravenously injection during operation and oxytocin 10 U intramuscular injection per 12 hours 3 times after CS. The following clinical parameter were collected and analyzed: (1) The amount of blood loss during operation, at 2, 6, 12, 24, 48 hours after operation. (2) The total amount of blood loss in 24 hours after CS and the incidence of postpartum hemorrhage. (3) The change of level of hemoglobin (Hb) and counting of red blood cell (RBC) from prepartum to postpartum. (4) Adverse reaction. RESULTS: (1) The mean amount of blood loss during operation were (368+/-258) ml in group of motherwort, (255+/-114) ml in group of motherwort+oxytocin and (269+/-141) ml in group of oxytocin, which exhibited significant difference among three groups (P<0.01). Meanwhile, no statistical different amount of blood loss among three groups were observed at 2, 6, 12, 24, 48 hours after CS. (2) The amount of blood loss of postpartum at 24 hours were (480+/-276) ml in group of motherwort, (361+/-179) ml in group of motherwort+oxytocin, (381+/-179) ml in group of oxytocin, which showed significant difference among 3 groups (P<0.01). (3) The incidence of postpartum hemorrhage were 32.0% (47/147) in group of motherwort, 11.1% (16/144) in group of motherwort+oxytocin, and 18.8% in (28/149) in group of oxytocin. When comparing the lowest rate of postpartum blood loss in group of motherwort+oxytocin and the highest rate in group of motherwort, it displayed statistical difference (P<0.01). (4) The decreased level of RBC and Hb were shown that RBC (0.3+/-0.5)x10(12)/L and Hb (9+/-13) g/L in group of motherwort, RBC (0.2+/-0.4)x10(12)/L and Hb (6+/-10) g/L in group of motherwort+oxytocin and RBC (0.2+/-0.4)x10(12)/L and Hb (7+/-30) g/L in group of oxytocin respectively. However, the comparison of different value of RBC and Hb in group of oxytocin and motherwort+oxytocin showed significant difference (P<0.05). (5) Two cases with allery reaction was observed. CONCLUSION: It is safe and efficacious that combined use of motherwort injection and oxytocin was to prevent postpartum hemorrhage during or after caesarian section.
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Cesárea , Medicamentos de Ervas Chinesas/uso terapêutico , Leonurus/química , Ocitocina/uso terapêutico , Hemorragia Pós-Parto/prevenção & controle , Adulto , Quimioterapia Combinada , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/efeitos adversos , Contagem de Eritrócitos , Feminino , Hemoglobinas/análise , Humanos , Injeções , Injeções Intramusculares , Ocitocina/administração & dosagem , Fitoterapia , Gravidez , Estudos Prospectivos , Método Simples-Cego , Resultado do Tratamento , Contração Uterina/efeitos dos fármacosRESUMO
OBJECTIVE: To study the effect of apoptosis induction of pianzihuang in human osteosarcoma U-20S cells. METHODS: Sixth SD rats were randomly divided into group A (8 rats) and group B (8 rats) in the study. Among them, 8 rats (group A) were treated with PBS and 8 rats (group B) treated with pianzihuang. (2) Human osteosarcoma U-2OS cells were cultured in nutrient solution (including fetal calf serum of 10%, penicillin of 100 IU/ml, streptomycin of 100 microg/ml), the environmental condition in saturated humidity 5% CO2, 37 degrees C. The osteosarcoma U-2OS cell proliferation effect by different saturation of blood serum was measured by MTT method. And the optimal concentration of the serum (including Pianzihuang) could be obtained. (3) The morphous of osteosarcoma U-2OS cell were observed by inverted phase contrast microscope and electron microscope. (4) Apoptosis cell was detected by TUNEL in situ end labeling. (5) Extracted DNA,agarose gel electrophoresis (AGE) to detect apoptosis cell by transmission electron microscope (TEM). RESULTS: (1) It was best that osteosarcoma U-2OS cell proliferation effect by 20% of the concentration of the serum (including Pianzihuang). (2) On inverted phase contrast microscope showed that the cells of group A were fusiform adherence growth; the cyton transform shrind and circle in 20% of the concentration of the serum (including Pianzihuang). Follow lasting time,the cell turning circle gradually increased. Necrosis cell and fragment were found by microscope. (3) The detected result of apoptosis cell in 20% of the concentration of the serum(including Pianzihuang) was positive by TUNEL assays. (4) AGE showed "ladder" strand of DNA, a special phenomenon of cellular apoptosis. (5) Chromatin aggregated along corrugativus caryotheca and apoptotic body appeared on cell surface by observation of TEM. CONCLUSION: Pianzihuang can inhibit proliferation and induce apoptosis for osteosarcoma U-2OS cell.
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Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , DNA/metabolismo , Relação Dose-Resposta a Droga , Humanos , Marcação In Situ das Extremidades Cortadas , Microscopia de Contraste de Fase , Osteossarcoma/patologiaRESUMO
In this article, the poly-calcon carboxylic acid (poly-CCA) film modified electrode was prepared by cyclic voltammetry (CV). Then, an electrochemical DNA biosensor was developed for detection of PML/RARA fusion gene in acute promyelocytic leukemia (APL) by using 18-mer single-stranded deoxyribonucleic acid as the capture probe. The capture probe was covalently attached through free amines on the DNA bases using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydrosulfosuccinimide (NHS) cross-linking reaction on a carboxylate-terminated poly-CCA monolayer modified glassy carbon electrode (GCE). The covalent immobilized capture probe could selectively hybridize with its target DNA to form double-stranded DNA (dsDNA) on GCE surface. The aim of this work is to provide a well-defined recognition interface for the detection of DNA. Differential pulse voltammetry (DPV) was used to monitor the hybridization reaction on the capture probe electrode. The decrease of the peak current of methylene blue (MB), an electroactive indicator, was observed upon hybridization of the probe with the target DNA. The results indicated that in pH 7.0 phosphate buffer solution (PBS), the oxidation peak current was linear with the concentration of complementary strand in the range of 1.0 x 10(-12) to 1.0 x 10(-11)M with a detection limit of 6.7 x 10(-13)M. This new method demonstrates its excellent specificity for single-base mismatch and complementary sequence (dsDNA) after hybridization, and it would be proposed to use in real sample.
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Técnicas Biossensoriais/métodos , Genes Neoplásicos , Proteínas de Fusão Oncogênica/genética , Animais , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/normas , Sondas de DNA , Técnicas Eletroquímicas , Eletrodos , Humanos , Leucemia Promielocítica Aguda/diagnóstico , Hibridização de Ácido NucleicoRESUMO
OBJECTIVE: To observe the effect of pilose antler polypeptides (PAP) on the apoptosis of rabbit marrow mesenchymal stem cells (MSCs) differentiated into chondrogenic phenotype by interleukin 1beta (IL-1beta) so as to optimize the seeding cells in cartilage tissue engineering. METHODS: The MSCs were separated from the nucleated cells fraction of autologous bone marrow by density gradient centrifuge and cultured in vitro. The MSCs were induced into chondrogenic phenotype by transforming growth factor beta1 (TGF-beta1) and basic fibroblast growth factor (bFGF). According to different medias, the MSCs were randomly divided into four groups: group A as black control group, group B (100 ng IL-1beta), group C (10 microg/ml PAP + 100 ng IL-1beta) and group D (100 ng/ml TGF-beta1 + 100 ng IL-1beta). The samples were harvested and observed by morphology, flow cytometry analysis, RT-PCR and ELISA at 24, 48 and 72 hours. RESULTS: The intranuclear chromatin agglutinated into lump and located under nuclear membranes which changed into irregular shape at 24 hours. The intranuclear chromatin agglutinated intensified at 48 hours. Then the nuclear fragments agglutinated into apoptosis corpuscles at 72 hours in group B. The structure change of cells in groups C and D was later than that in group B, and the number of cells changed shape was fewer than that in group B. The structure change of cells in group A was not significant. The apoptosis rate of cells, the mRNA expression of Caspase-3 and the enzymatic activity of Caspase-3 gradually increased in group B, and there were significant differences compared with groups A, C and D (P < 0.01). CONCLUSION: Caspase-3 is involved in apoptosis of the MSCs differentiated into chondrogenic phenotype cultured in vitro. PAP could prevent from or reverse apoptosis of these MSCs by decreasing the expression of Caspase-3 and inhibiting the activity of Caspase-3.