OBJECTIVES: To investigate the role of tetramethylpyrazine(TMP) nitrone in proliferation and differentiation of neural stem cells (NSCs). METHODS: We separated and cultivated the original generation of NSCs from cerebral cortex of 14 days rat embryo, and the phenotype characteristics of the third-generation NSCs was tested by immunofluorescence. The experiment was divided into control group, β-mercaptoethanol positive control group, tetramethylpyrazine nitrone group and tetramethylpyrazine nitrone + ethylene glycol tetraacetic acid(EGTA) group (=4). The third-generation cultivation of NSCs was used in the experiment. The effect of tetramethylpyrazine nitrone on the number of NSCs proliferation was determined by BrdU and MTT, and the differentiation of NSCs was determined by Western blot. RESULTS: The primary NSCs was isolated successfully, neurospheres with typical NSCs morphology and expressing nestin was formed at 3-5 days. As BrdU and MTT assay results shown, compared with the control group andβ-mercaptoethanol positive control group, the NSCs proliferation numbers of tetramethylpyrazine nitrone group increased significantly(<0.05). The results of Western blot showed that the neuronal differentiation rate of NSCs was increased significantly in both the tetramethylpyrazine nitrone group and tetramethylpyrazine nitrone + EGTA group, and the differentiation rate of NSCs in tetramethylpyrazine nitrone + EGTA group increased more significantly(<0.05). CONCLUSIONS Tetramethylpyrazine nitrone can significantly enhance the proliferation and neuronal differentiation rate of NSCs. Decrease in extracellular Ca can promote the differentiation of NSCs into neurons induced by tetramethylpyrazine nitrone. Ca signaling plays an important role in the differentiation of NSCs into neurons.
Animals ; Calcium Signaling ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Neural Stem Cells ; cytology ; drug effects ; Nitrogen Oxides ; pharmacology ; Pyrazines ; pharmacology ; Rats

Humans ; Methylene Blue ; Melanoma ; Staining and Labeling

To investigate the effects of saponins extracted from Panax japonicus(SPJ) on cardiomyocyte apoptosis in natural aging rats and explore its underlying mechanisms. SD male rats were randomly divided into four groups: young control group, natural aging group, SPJ low dose group and SPJ high dose group, with 10 rats in each group. The rats in natural aging group, SPJ low and high dose groups were respectively treated with normal saline, SPJ 10 and 60 mg•kg-1•d-1 from the beginning of 18 month-old, 6 days per week for 6 months till 24 month-old. Then the animals were sacrificed. Their myocardial morphology changes were observed by using haematoxylin-eoin(HE) staining; cardiomyocyte apoptosis was tested by using Tunel assays; and the protein expression levels of Bcl-2, Bax, IL-1β, TNF-α, AMPK, p-AMPK, Sirt1, and Ac-NF-κB p65 in myocardial tissues of rats were detected by Western blot. The results showed that SPJ could effectively improve the arrangement disorder of myocardial fibers, reduce the infiltration of inflammatory cells and inhibit cardiomyocyte apoptosis in natural aging rats. At the same time, SPJ could significantly inhibit the protein expression of Bax, IL-1β, TNF-α and Ac-NF-κB p65, and increase the expression of Bcl-2, Bcl-2/Bax, p-AMPK/AMPK and Sirt1 in the heart tissues of natural aging rats. SPJ can effectively inhibit cardiomyocyte apoptosis in natural aging rats, and its mechanisms may be related with the regulation of inflammatory reaction by AMPK/Sirt1/NF-κB signaling pathway.

To investigate the amelioration effect of saponins extracted from Panax japonicas (SPJ) on myocardial fibrosis in natural aging rats and its mechanisms, male SD rats aged 18 months were randomly divided into 3 groups (aging model group, low-dose SPJ group and high-dose SPJ group), with 10 rats in each group. SPJ groups were given SPJ at different doses (10, 60 mg·kg⁻¹·d⁻¹) consecutively for 6 months, meanwhile, aging model group was treated with the equal volume of saline for 6 months until 24 months old. Another 10 rats aged 6 month were used as young control group. The changes of myocardial morphological were observed by haematoxylin-eosin (HE) staining. Masson staining was used to observe the changes of collagen deposition in rat hearts. RT-PCR was used to detect the mRNA expression levels of myofibroblast marker α-SMA, collagen-related protein COL1α2, COL3α1 and matrix metalloproteinase MMP2, MMP9. Western blot was used to test the changes of the protein expressions of TGF-β1, p-Smad3, IL-1β and TNF-α in heart tissues. SPJ can effectively improve the arrangement of myocardial fibers, decrease inflammatory infiltration and reduce collagen deposition in aging rats. SPJ can effectively down-regulate the mRNA expression levels of COL1α2, COL3α1, α-SMA, MMP9, MMP2 and inhibit the protein expressions of TGF-β1, p-Smad3, TNF-α, IL-1β in the natural aging heart tissues. SPJ can effectively alleviate myocardial fibrosis in natural aging rats, and its mechanisms was related to the inhibition of the protein expressions of TGF-β1, p-Smad3 and the reduction of myocardial inflammation in rat hearts.
Animals ; Fibrosis ; Male ; Panax ; Rats ; Rats, Sprague-Dawley ; Saponins ; Signal Transduction ; Smad3 Protein ; Transforming Growth Factor beta1

Osteosarcoma is the most common malignant tumors of bone. Since 1970s, researchers had used chemotherapy drugs to treat osteosarcoma. However, multidrug resistance is a major adverse reaction that affects the efficacy of chemotherapy drugs, leading to the reduced survival rate of osteosarcoma patients. The Notch signaling pathway plays an important role in osteosarcoma proliferation, which affects tumor resistance by reducing intracellular drug accumulation, regulating epithelial-mesenchymal transition, dysregulating microRNA, disrupting the expression of apoptosis genes, and regulating tumor stem cells.
Bone Neoplasms/drug therapy* ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Osteosarcoma/drug therapy* ; Pharmaceutical Preparations ; Receptors, Notch/genetics* ; Signal Transduction

Programmed cell death(PCD)is a unique cell death involving effector molecules, including various forms such as apoptosis, autophagy, and pyroptosis. PCD is involved in many aspects of normal physiological activities in humans, and is closely related to the development of many diseases. Glaucoma is the leading cause of irreversible blindness worldwide. Relevant studies have shown that the development of glaucoma is associated with the abnormal expression of a variety of PCD-related proteins. The mechanism and interplay of apoptosis, autophagy, pyroptosis, ferroptosis and parthanatos of retinal ganglion cells in the course of glaucoma were reviewed, to provide a new direction for the prevention treatment of glaucoma.

The angle between the lower extremity force line and the position of the unicondylar prosthesis is an important factor affecting the long-term survival and rate clinical outcome of the unicondylar replacement prostheses. Insufficient lower limb alignment will accelerate the wear of prosthesis and reduce the survival rate of prosthesis. Excessive lower limb alignment will accelerate the progress of contralateral interventricular arthritis. It is generally believed that the lower limb force line should be corrected in mild varus after unicompartmental knee arthroplasty. However, some scholars believe that the lower limb alignment has no effect on the functional score and prosthesis survival rate after unicompartmental knee arthroplasty. The poor position of femoral and tibial prosthesis will cause unexplained pain and even prosthesis wear, but the optimal position of femoral and tibial prosthesis is controversial. It is generally believed that the posterior tibial slope should be corrected in the range of 3° to 7° in unicompartmental knee arthroplasty, but some scholars believe that excessive change of posterior tibial slope will also affect the balance of knee joint space and knee joint range of motion. This study shows that the correction of lower limb alignment to mild varus is still the best lower limb alignment for unicompartmental knee arthroplasty. The best position of femoral and tibial prosthesis needs to be confirmed by further biomechanical research. The correction of tposterior tibial slope should be changed according to the specific original dissection angle of patients before operation.
Humans ; Arthroplasty, Replacement, Knee ; Knee Prosthesis ; Knee Joint/surgery* ; Tibia/surgery* ; Osteoarthritis, Knee/surgery* ; Lower Extremity/surgery* ; Retrospective Studies

OBJECTIVE: To investigate the clinical effect the treatment of arthroscopy-assisted calcaneal spur resection combined with plantar fascia release and calcaneal decompression in the treatment of the patients with intractable calcaneal pain. METHODS: The clinical data of 50 patients with intractable heel pain from January 2016 to January 2019 were retrospectively analyzed, including 20 males and 30 females;aged from 40 to 68 years old with an average of (50.12±7.35)years old, the medical history ranged from 1 to 4 years. All patients underwent arthroscopy-assisted calcaneal spur resection combined with plantar fascia release and calcaneal decompression, and were followed up, the duration ranged from 24 to 60 months with an average of(42.00±3.28) months. All patients had obvious heel pain before surgery, and X-ray examinations often showed the presence of calcaneal spurs. In addition to the routine foot examination, the changes in the height and angle of the arch of the foot were also measured pre and post-operatively by X-ray, for the evaluation of clinical effect. The VAS system was used to evaluate the degree of foot pain;the AOFAS scoring system was used to comprehensively evaluate the foot pain, voluntary movement, gait and stability. RESULTS: The VAS decreased from (8.75±1.24) before surgery to (5.15±2.35) at 3 months after surgery, (4.07±2.53) at 6 months after surgery, and (3.95±2.44) at the last fllow-up(P<0.05). The AOFAS score increased from (53.46±4.17) before surgery to(92.46±2.53) at 3 months after surgery, (96.33±2.46) at 6 months after surgery, and (97.05±2.37) at the last follow-up(P<0.05). The arch height was (41.54±1.15) mm before operation and (41.49±1.09) mm after the operation, the difference was not statistically significant(P>0.05). The internal arch angle of the foot arch was (121±6)° before operation and (122±7)° after operation. The difference was not statistically significant(P>0.05). CONCLUSION Arthroscopy-assisted calcaneal bone spurs resection combined with plantar fascia release and calcaneal decompression exhibited great clinical effect for treating intractable heel.
Male ; Female ; Humans ; Adult ; Middle Aged ; Aged ; Heel/surgery* ; Heel Spur/surgery* ; Retrospective Studies ; Calcaneus/surgery* ; Foot Diseases ; Pain ; Endoscopes ; Treatment Outcome

OBJECTIVETo observe the action of Chinese herbal preparation (CHP) on the recovery of nasal sinus mucosa after endoscopic sinus surgery (ESS).

METHODSSeventy-eight patients (140 sinus) with chronic nasosinusitis and nasal polyps, who had undergone ESS, were divided into two groups. The 40 patients (70 sinuses) in the experiment group were treated with daily nasal flushing with diluted CHP while the 38 patients (70 sinuses) in the control group were untreated. Tissues of nasal mucosa were taken out from patients' posterior walls of maxillary sinus at different time points, i. e. 2 - 3 weeks, 8 - 11 weeks and 13 - 15 weeks after ESS were observed.

RESULTSSignificant difference was shown between the two groups under endoscope at all the time points in occurrence (patient/sinus) of sticky mucus, swelling and thickened mucus, occlusion of sinus opening, bloody secretion in sinus, and adhesion, etc. (P < 0.05, P < 0.01). Light microscopic examination showed significant difference between the two groups in occurrence (patient/sinus) of squamous epithelial metaplasia and fiber tissues proliferation (P <0.05). And electron microscopic examination also showed significant difference between the two groups in occurrence (patient/sinus) of inflammatory cell infiltration, fibers decrease, disordered arrangement, microvilli and short cilia (P<0.01).

CONCLUSIONUsing CHP for postoperative nasal flushing after ESS is a safe and manageable approach with effects in promoting nasal mucosa recovery.

Adolescent ; Adult ; Aged ; Convalescence ; Drugs, Chinese Herbal ; administration & dosage ; Endoscopy ; Female ; Humans ; Male ; Middle Aged ; Nasal Mucosa ; pathology ; Nasal Polyps ; complications ; drug therapy ; surgery ; Otorhinolaryngologic Surgical Procedures ; Postoperative Period ; Sinusitis ; complications ; drug therapy ; surgery

OBJECTIVETo investigate the correlation between methylation status and gene expression of APC (adenomatous polyposis coli) gene in HeLa, CaSki and SiHa cell lines of cervical carcinoma, and explore the effect of hydralazine on the transcription regulation of the 5'CpG island demethylation of APC gene and the proliferation and apoptosis of the cell lines.

METHODSMethylation status and the expression of APC gene were analyzed using methylated specific PCR, RT-PCR and FQ-PCR methods. The expression of beta-catenin protein which correlates closely with APC was detected by SP method after treatment with Hydralazine. MTT and FCM assays were used to observe the changes of proliferation activity and apoptosis of the cells after Hydralazine treatment.

RESULTS(1) APC gene was methylated or hemimethylated respectively in HeLa and CaSki cell lines, at the same time, APC gene was not methylated in SiHa cell. (2) After having been treated by 40 micromol/L Hydralazine for 72 hours, growth inhibitory ratios of HeLa, CaSki and SiHa cell lines were (52.12 +/- 3.78)%, (44.31 +/- 2.59)% and (47.73 +/- 4.73)% respectively, on the contrary, normal cell HECV's growth inhibitory ratio was only (27.18 +/- 0.79)%. APC gene in HeLa and CaSki cell lines which were treated by 40 micromol/L Hydralazine for 72 hours was demethylated and expressed positively, the expression of APC mRNA in HeLa, CaSki and SiHa cell lines increased to 10.35, 11.40 and 0.73 times respectively. (3) Hydralazine, when used at the concentration of 40 micromol/L for 72 hours, induced S phase and G2/M phase arrest and apoptosis in HeLa and CaSki cells. beta-catenin protein can be expressed in cell membrane after treatment with Hydralazine.

CONCLUSIONAPC gene methylation plays an important role in the carcinogenesis of cervical cells and can re-express after the treatment with Hydralazine which also could inhibit the growth of the cervical cancer cells.

Adenomatous Polyposis Coli Protein ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; CpG Islands ; genetics ; DNA Methylation ; Female ; Gene Expression Regulation, Neoplastic ; Genes, APC ; HeLa Cells ; Humans ; Hydralazine ; pharmacology ; RNA, Messenger ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology ; beta Catenin ; metabolism