Objective To study the relationship of insulin receptor substrate-1 (Irs1) and metabolic disease, we generated Irs1 gene knockout rat by CRISPR/Cas9 system.Methods Two sgRNA targeting sites were designed for Irs1 targeting.The Cas9 and sgRNAs were transcribed by T7 RNA polymerase in vitro.Cas9 mRNA and sgRNA mixtures were pooled and microinjected into one-cell fertilized eggs of SD rats to generate rats with targeted mutation .Results Five rats with the mutations were detected with the efficiency of 83%.Conclusion The Irs1 gene knockout rats generated in this study can be transmitted by germline .

This study examined the ability of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (β-PGG) to induce the expression of heme oxygenase-1 (HO-1) in the PC12 cells and its regulation in the PC12 cells. One week before treatment with the drug, nerve growth factor (NGF) was added to the cultures at a final concentration of 50 ng/mL to induce neuronal differentiation. After drug treatment, HO-1 gene transcription was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Expression of HO-1 and NF-E2-related factor2 (Nrf2) and activation of extracellular signal-regulated kinase (ERK) and Akt were detected by Western blotting. The viability of the PC12 cells treated with different medicines was examined by MTT assay. The oxidative stress in the PC12 cells was evaluated qualitatively and quantitatively by DCFH-DA. The results showed that β-PGG up-regulated HO-1 expression and this increased expression provided neuroprotection against MPP(+)-induced oxidative injury. Moreover, β-PGG induced Nrf2 nuclear translocation, which was found to be upstream of β-PGG-induced HO-1 expression, and the activation of ERK and Akt, a pathway that is involved in β-PGG-induced Nrf2 nuclear translocation, HO-1 expression and neuroprotection. In conclusion, β-PGG up-regulates HO-1 expression by stimulating Nrf2 nuclear translocation in an ERK- and Akt-dependent manner, and HO-1 expression by β-PGG may provide the PC12 cells with an acquired antioxidant defense capacity to survive the oxidative stress.

Objective To knock out the Abcb1 gene of rat,and establish the Abcb1 humanized rat model based on the Abcb1 knock out rat.Methods The animal model was established using BAC and CRISPR/Cas9 technology,and was analyzed by PCR, RT-PCR and real-time PCR.Results Establishing a rat model expressing human Abcb1 stably by transfer the 153 kb BAC containing human Abcb1 promoter and cDNA into rat genome, and establishing the Abcb1 knock out rat at the same time.Establishing the Abcb1 humanized model by crossing these two strains together.The expression pattern of Abcb1 in Abcb1 humanized rat is different from the wild type rat.The Abcb1 humanized model express not only the human Abcb1 gene but also has similar expression pattern as human.Conclusions The Abcb1 knock out rat and the Abcb1 humanized rat were successfully established, and this model is close to human concerning about the drug metabolism related to Abcb1.

To explore the relationship between beta-amyloid (A beta) and the pathogenesis of Alzheimer disease (AD), after injection of beta-amyloid into the rat brain, the apoptosis of nerve cells and acetylcholine (Ach) content in rat hippocampus were examined by employing TUNEL technique and base hydroxylamine colorimetry respectively. The influence of age and glucocorticoid on the neurotoxic effect of A beta was also analyzed. A beta peptide could strongly induce the apoptosis of neurons in hippocampus, cortex and striate body (P < 0.05 or P < 0.01). In addition, the senility and glucocorticoid pre-treatment could enhance the toxic effect of A beta (P < 0.05 or P < 0.01). It is concluded that A beta may play an important role in the pathogenesis of Alzheimer disease via its induction of apoptosis of neurons and by decreasing the content of the Ach.
Acetylcholine/metabolism ; Aging ; Alzheimer Disease/etiology ; Amyloid beta-Protein/*toxicity ; Apoptosis/*drug effects ; Dexamethasone/*pharmacology ; Drug Synergism ; Hippocampus/metabolism ; Hippocampus/*pathology ; Injections, Intraventricular ; Neurons/pathology ; Rats, Wistar

Objective:To evaluate the quality of prostate hyperplasia related literature in acupuncture and moxibustion,and to compare the curative effect on prostate hyperplasia between acupuncture and moxibustion and western medicine.Methods:Retrieving Pubmed,Cochrane Library,CBM database,CNKI database Etc.to collect the literature of prostate hyperplasia of clinical randomized or quasi-randomized control trials of comparative study between western medicine and acupuncture treatment.The data was extracted independently by two valuers from literatures fitting the selection criteria.Cochrane evaluation manual 4.2.6 was used to evaluate quality,and RevMan 4.2.8 was used in statistical analysis.Results:A total of six randomized or quasi-randomized controlled trials (total 546 examples) were adopted.6 study adopted the total effective rate of evaluation indexes,Meta-analysis showed that there was a significant difference between acupuncture treatment group and western medicine group [merger RR (fixed effects model)=1.26,95%CI(1.15,1.37),Z=5.13,P

Regulation of the adhesion molecules expression by cytokine in vascular endothelial cells was investigated. Human umbilical vein endothelial cells (HUVEC) were stimulated with cytokines, TNF-α (1-250 U/ml) or IL-1β (0.1-50 U/ml) for 24 h. HUVEC were also cultured with cytokines, TNF-α (100 U/ml) or IL-1β (10 U/ml), for 4-72 h, cell surface expression of adhesion molecules (ICAM-1 and VCAM-1) were detected and quantitated by immunocytochemical methods and computerized imaging analysis technique. Adhesion molecules expression were up-regulated by TNF-α, IL-1β in a concentration- and time-dependent manner. Some significant differences were observed between the effects of cytokines on the ICAM-1 and on VCAM-1 expression. Cytokines might directly induce the expression of ICAM-1 and VCAM-1 in vascular endothelial cells. Our observations indicate differential functions of the two adhesion molecules during the evolution of inflammatory responses in stroke.

Regulation of the adhesion molecules expression by cytokine in vascular endothelial cells was investigated. Human umbilical vein endothelial cells (HUVEC) were stimulated with cytokines, TNF-α (1-250 U/ml) or IL-1β (0.1-50 U/ml) for 24 h. HUVEC were also cultured with cytokines, TNF-α (100 U/ml) or IL-1β (10 U/ml), for 4-72 h, cell surface expression of adhesion molecules (ICAM-1 and VCAM-1) were detected and quantitated by immunocytochemical methods and computerized imaging analysis technique. Adhesion molecules expression were up-regulated by TNF-α, IL-1β in a concentration- and time-dependent manner. Some significant differences were observed between the effects of cytokines on the ICAM-1 and on VCAM-1 expression. Cytokines might directly induce the expression of ICAM-1 and VCAM-1 in vascular endothelial cells. Our observations indicate differential functions of the two adhesion molecules during the evolution of inflammatory responses in stroke.

This study examined the ability of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (β-PGG) to induce the expression of heme oxygenase-1 (HO-1) in the PC12 cells and its regulation in the PC12 cells. One week before treatment with the drug, nerve growth factor (NGF) was added to the cultures at a final concentration of 50 ng/mL to induce neuronal differentiation. After drug treatment, HO-1 gene transcription was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Expression of HO-1 and NF-E2-related factor2 (Nrf2) and activation of extracellular signal-regulated kinase (ERK) and Akt were detected by Western blotting. The viability of the PC12 cells treated with different medicines was examined by MTT assay. The oxidative stress in the PC12 cells was evaluated qualitatively and quantitatively by DCFH-DA. The results showed that β-PGG up-regulated HO-1 expression and this increased expression provided neuroprotection against MPP(+)-induced oxidative injury. Moreover, β-PGG induced Nrf2 nuclear translocation, which was found to be upstream of β-PGG-induced HO-1 expression, and the activation of ERK and Akt, a pathway that is involved in β-PGG-induced Nrf2 nuclear translocation, HO-1 expression and neuroprotection. In conclusion, β-PGG up-regulates HO-1 expression by stimulating Nrf2 nuclear translocation in an ERK- and Akt-dependent manner, and HO-1 expression by β-PGG may provide the PC12 cells with an acquired antioxidant defense capacity to survive the oxidative stress.
Animals ; Cell Death ; drug effects ; genetics ; Cell Line, Tumor ; Heme Oxygenase-1 ; genetics ; Hydrolyzable Tannins ; pharmacology ; MAP Kinase Signaling System ; drug effects ; genetics ; PC12 Cells ; Piperidines ; adverse effects ; Proto-Oncogene Proteins c-akt ; genetics ; Pyrazoles ; adverse effects ; Rats

To explore the relationship between beta-amyloid (A beta) and the pathogenesis of Alzheimer disease (AD), after injection of beta-amyloid into the rat brain, the apoptosis of nerve cells and acetylcholine (Ach) content in rat hippocampus were examined by employing TUNEL technique and base hydroxylamine colorimetry respectively. The influence of age and glucocorticoid on the neurotoxic effect of A beta was also analyzed. A beta peptide could strongly induce the apoptosis of neurons in hippocampus, cortex and striate body (P < 0.05 or P < 0.01). In addition, the senility and glucocorticoid pre-treatment could enhance the toxic effect of A beta (P < 0.05 or P < 0.01). It is concluded that A beta may play an important role in the pathogenesis of Alzheimer disease via its induction of apoptosis of neurons and by decreasing the content of the Ach.
Acetylcholine ; metabolism ; Aging ; Alzheimer Disease ; etiology ; Amyloid beta-Peptides ; toxicity ; Animals ; Apoptosis ; drug effects ; Dexamethasone ; pharmacology ; Drug Synergism ; Hippocampus ; metabolism ; pathology ; Injections, Intraventricular ; Male ; Neurons ; pathology ; Rats ; Rats, Wistar

Objective To evaluate the clinical efficacy and safety of short-segment pedicle screw fixation combined with vertebroplasty for the treatment of thoracolumbar osteoporotic compression fractures.Methods A retrospective case control study was conducted on 63 patients with fresh thoracic or lumbar osteoporotie compression fractures who were surgically treated from January 2010 to December 2013.There were 26 males and 37 females,with age of 63-87 years [(76.3 ± 5.7) years].According to the surgical method,the patients were assigned to simple vertebroplasty group (Group A),short-segment pedicle screw fixation group (Group B),and short-segment pedicle screw fixation group combined with vertebroplasty group (Group C),with 21 cases in each group.Length of hospital stay,operation time,and blood loss were recorded.The visual analog scale (VAS),anterior vertebral body height,angle of the kyphotic deformity,and complications before operation,immediately after operation,3 months after operation,and at the last follow up were compared among three groups.Complications of each group were recorded.Results The hospital stay,operation time,and blood loss in Group C were significantly higher than those in Group A (P ＜ 0.05),but there were no significant differences between Group B and Group C (P ＞ 0.05),except for a longer operation time in Group C.The pre-operative VAS showed no significant difference among three groups (P ＞ 0.05).However,the mean VAS in Groups A,B and C at the last follow up were 1.0(0.0,2.0)points,1.0(0.0,2.0)points,0.0(0.0,1.0)points,respectively.The VAS in Group C was significantly lower than that in Group B or A (P ＜ 0.05).The mean anterior vertebral body height and angle of the kyphotic deformity in Group C had no significant difference from that in Group A or B before operation and immediately after operation (P ＞ 0.05).At the last follow up,the mean anterior vertebral body height and angle of the kyphotic deformity in Groups A,B and C were (68 ±14.7)％,(72.3 ±9.0)％,(81.5 ±5.6)％ and (10.6 ±3.9)°,(10.7 ±5.0)°,(7.4 ± 5.0) °,respectively.The loss of anterior vertebral body height and angle of the kyphotic deformity correction in Group C were significantly less than that in Group A or B (P ＜ 0.05).Superficial infection was found in Groups B (n =2) and C (n =1),and the infection was cured after antibiotic therapy and dress change.Bone cement leakage was found in Groups A (n =8) and C (n =5),with no nerve compression.Internal fixation failure was seen in Group B (n =3),and the implant was removed directly.Conclusion Short-segment pedicle screw fixation combined with vertebroplasty can effectively reduce the loss of anterior vertebral body height and angle of the kyphotic deformity and hence enhance the clinical efficacy.