OBJECTIVE To explore the effect and potential mechanism of the compatibility of Astragali Radix-Puerariae Lobatae Radix on ferroptosis of liver cells in type 2 diabetes mellitus （T2DM） insulin resistance （IR） rats. METHODS Sixty male SD rats were randomly divided into control group （12 rats） and modeling group （48 rats）. The modeling group was fed with a high- fat diet for 4 consecutive weeks and then given a one-time tail vein injection of 1% streptozotocin to establish T2DM IR model. The model rats were randomly divided into model group， the compatibility of Astragali Radix-Puerariae Lobatae Radix group [QG group， 4.05 g/（kg·d）， intragastric administration]， ferroptosis inhibitor ferrostatin-1 group [Fer-1 group， 5 mg/kg by intraperitoneal injection， once every other day]， the compatibility of Astragali Radix-Puerariae Lobatae Radix+ferroptosis inducer erastin group [QG+erastin group， 4.05 g/（kg·d） by intragastric administration+erastin 10 mg/（kg·d）， intraperitoneal injection]. After 4 weeks of intervention， serum fasting blood glucose （FBG） and fasting insulin （FINS） were measured in each group of rats， and homeostasis model assessment of insulin resistance （HOMA-IR） and the natural logarithm of insulin action index（IAI） were calculated； the serum levels of total cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate transaminase （AST） and alanine transaminase （ALT）， Fe2+ and Fe content， glutathione （GSH）， malondialdehyde （MDA） and superoxide dismutase （SOD） levels， NADP+/NADPH ratio and reactive oxygen species （ROS） were determined. The pathological morphology of its liver tissue was observed； the protein expressions of glutathione peroxidase 4 （GPX4）， ferritin heavy chain 1 （FTH1）， long-chain acyl-CoA synthetase 3 （ACSL3）， ACSL4， ferritin mitochondrial （FTMT）， and cystine/glutamate anti-porter （xCT） in the liver tissue of rats were detected. RESULTS Compared with control group， the liver cells in the model group of rats showed disordered arrangement， swelling， deepened nuclear staining， and more infiltration of inflammatory cells， as well as a large number of hepatocyte vacuoles and steatosis； FBG （after medication）， the levels of TC， TG， LDL-C， AST， ALT， FINS， MDA and ROS， HOMA-IR， Fe2+ and Fe content， NADP+/NADPH ratio and protein expression of ACSL4 were significantly increased or up-regulated， while the levels of HDL-C， GSH and SOD， IAI， protein expressions of GPX4， FTH1， ACSL3， FTMT and xCT were significantly reduced or down-regulated （P＜0.01）. Compared with the model group， both QG group and Fer-1 group showed varying degrees of improvement in pathological damage of liver tissue and the levels of the above indicators， the differences in the changes of most indicators were statistically significant （P＜0.01 or P＜0.05）. Compared with QG group， the improvement of the above indexes of QG+erastin group had been reversed significantly （P＜0.01）. CONCLUSIONS The compatibility decoction of Astragali Radix-Puerariae Lobatae Radix can reduce the level of FBG in T2DM IR rats， and alleviate IR degree， ion overload and pathological damage of liver tissue. The above effects are related to the inhibition of ferroptosis.

ObjectiveTo perform pharmacology problem-based learning (PBL) and evaluate its effects.MethodsPBL was performed for the clinical medicine class of grade 2007 and the satisfactory degree of students to teaching effects was observed with questionnaire. Results The students thought that PBL teaching had substantial contents and proper schedule and increased learning interest. Students' participating degree, mutual communication and controlling discussion procedure were fine,which reached the expected learning objective. ConclusionsThe effects of PBL teaching were excellent and most of our students could accept it.

BACKGROUND:Intensity-time (I/T) curve examination is a monitoring technology used for diagnosis of nerve damage,muscle disease and prognostic evaluation using current stimulation and qualitative or quantitative analysis.It also has significance to the diagnosis of lumbar disc herniation (LDH).Whether it can be used in LDH prognostic evaluation is poorly understood.OBJECTIVE:To explore the effectiveness of I/T curve in diagnosis and prognostic evaluation of LDH.METHODS:I/T curves of 113 LDH patients were measured by CX-3 electrodiagnostic equipment,and the results were compared with their unaffected sides and analyzed after physiotherapy.A total of 253 curves were measured,including 107 biceps femoris,101 gastrocnemius and 45 tibialis anterior muscle curves.All patients were sequential treated by traction,medium frequency,ultrashort wave,electric acupuncture as well as infrared radiation following I/T diagnosis,once a day,10 days for a course,with 10 days interval in 3 courses,I/T curves were performed after 3 courses.The therapeutic effect was evaluated by using I/T results combined with clinical symptom and physical signs.RESULTS AND CONCLUSION:The first estimated percentage by I/T curved line were 36.36% (controlled by normal nerve),62.85% (controlled by part of the denervated),0.79% (controlled by the completely denervated) and 63.64% (controlled by total abnormal nerve).After the physiotherapy,the effectiveness of the complete recovery was 92.86% by the normal nerve as well as 58.82% by the abnormal nerve.Above all,the practical value could be concluded from the diagnosis,evaluation and effectiveness of LDH used by I/T curve line.The therapeutic effectiveness controlled by the normal nerve is much better than that controlled by the abnormal.

Objective To study the effects of electromagnetic field (EMF) exposure on osteoporosis in ovariectomized mice. Methods Sixty 8-week-old female Kunming mice were divided into four groups at random: a sham operation group (group A), an ovariectomized group (group B), an EMF and ovariectomized group (group C) and a nilestriol and ovariectomized group (group D). Bilateral ovariectomies were performed on all mice except those in group A. The mice of group C were exposured to a 15 Hz, 1.0 mT electromagnetic field. The mice of group D were given at nilestriol 1.5 mg/kg/week. The bone mineral density (BMD) of the lumbar vertebrae was measured before the mice were sacrificed at the 12th week. Blood specimens were collected every two weeks to measure the ac-tivity of alkaline phosphatase (ALP) and the concentration of bone gamma-carboxyglutamic-acid-containing proteins (BGP), calcium and estradiol in the serum. Histological sections were taken to examine and analyze the changes in bone trabeculae in the lumbar vertebrae after 6 and 12 weeks. Results EMF at 15 Hz and 1.0 mT intensity signifi-cantly increased the activity of ALP and the concentrations of BGP and calcium in the serum. In addition, the absorp-tion of bone trabeculae in the lumbar vertebrae was significantly restrained. Conclusions EMF at 15 Hz and 1.0 mT can restrain the development of osteoporosis in ovariectomized mice.

Objective To study the effects of an electromagnetic field (EMF) on the expression of fibro-blast growth factor (FGF-2) and it' s receptor (FGFR-2) mRNA in rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods Rat BMSCs were isolated and cultured in vitro. The subcultured cells were divided into different groups to be EMF stimulated at 1.0 mT. The expression of FGF-2 and FGFR-2 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results Different frequencies and durations of 1.0 mT EMF exposure induced FGF-2 and FGFR-2 mRNA expression in comparison to blank controls. The expression of FGF-2 mRNA reached a peak after stimulation at 15 Hz for 10 min, 50 Hz for 60 min and 75 Hz for 30 min. And the expression of FGFR-2 mRNA reached a peak after 30 minutes at all frequencies. At 1.0 mT with 30 min exposure, the expression of FGF-2 mRNA peaked after 50 Hz stimulation, and the expression of FGFR-2 mRNA peaked after stimulation at 75 Hz. Conclusions Moderate EMF stimulation can significantly increase the expression of FGF-2 and FGFR-2 mRNA in rat BMSCs in vitro.

Objective To study the effects of pulsed electromagnetic field (PEMF) in promoting tibia and fibula fracture healing in rats. Methods Thirty rats were divided randomly into two groups after establishing the animal model of artificial tibia and fibula fracture: a PEMF treated group and a control group. Radiographs were taken immediately postoperatively and once a week thereafter until being sacrificed after week 5. The blood was sampled to measure the activity of alkaline phosphatase (ALP) and the concentration of bone gamma-carboxyglutamic-acid-con-taining proteins (BGP), calcium and phosphate in serum once a week, respectively. Histological sections were taken at weeks 3 and 5 to observe the pathological change of bony callus. Results PEMF of 15 Hz and 1.0 mT could sig-nificantly increase the amount of bony callus, significantly increase the amount of bony callus, promote the disappea-ring of fracture lines and the appearance of endochondral ossification and mature bone trabecula. The amount of colla-gen in bony callus raised and the process of callus moulding accelerated in PEMF treated group. In addition, the ac-tivity of ALP(P<0.04) and the concentration of BGP(P<0.05) in serum increased. Conclusions The PEMF of 15 Hz and 1.0 mT can obviously promote fracture healing of tibia and fibula in rats.

Objective To study the expressions of aggrecan (Agc) Ⅰ and Ⅱ collagen and Sox9 by bone mar-row mesenchymal stem cells exposed to electromagnetic fields (EMFs) and it's mechanisms involved. Methods Bone marrow mesenchymal stem cells were isolated from Sprague-Dawley rats and cultured in vitro. The third passage cells were harvested and exposed to 15 Hz 1 mT EMFs for 8 h/d. The semi-quantitative reverse transcription-polyme-rase chain reaction (RT-PCR) technique was used to measure parathyroid hormon receptor related peptide (PTHrp) ,Agc Ⅰ and Ⅱ collagen and Sox9 mRNA. Western blotting was used to measure type Ⅱ collagen expression. After the inhibitor of protein kinase A (PKA) H-89 and the inhibitor of protein kinase C (PKC) Go-6976 ( 12 μm) were added, the effects of EMFs on Agc Ⅰ and Ⅱ collagen and Sox9 mRNA expressions were measured again by using RT-PCR, and Western blotting technique. Results The EMFs induced significant increase of mRNA expressions of PTHrp, Agc Ⅰ and Ⅱ collagen and Sox9 in comparison to the controls, and promoted type Ⅱ collagen protein expres-sion. The Agc Ⅰ and Ⅱ collagen expressions decreased after PKA pathway inhibitor H-89 and PKC inhibitor Go-6976 were added, but the mRNA expression of Sox9 was not affected. Conclusion This study shows 15Hz 1mT EMFs can promote mRNA expressions of Agc Ⅰ and Ⅱ collagan and Sox9 of cbondrogenesis differentiation markers in bone marrow mesenchymal stem cells. The effect is correlated with PKA and PKC pathways.

Objective To study the influence of pulsed electromagnetic fields on the expression of type I collagen by bone marrow mesenchymal stem cells and it's mechanism. Methods The bone marrow mesenchymal stem cells of SD rats were isolated and cultured in vitro.The third passage cells were harvested and exposed to pulsed electromagnetic fields(PEMFs)at 15 Hz and 1 mT 8 h/d for 3 days.A semi-quantitative RT-PCR technique was used to measure the type I collagen mRNA expression;ELISA and immunohistochemitistry techniques were used to measure type I collagen expression.Inhibitors and promoters of the cAMP-dependent protein kinase A(cAMP-PKA)pathway were added.After the cAMP-PKA pathway had been inhibited or promoted,the effects of the PEMF on type I collagen expression were measured again using ELISA and immunohistoehemistry.Results PEMFs at 15 Hz and 1 mT induced significant promotion of the expression of type I collagen(P≤0.01)in comparison with the controls. The type I collagen expression was reduced when the cAMP-PKA pathway inhibitor H-89 was added,and raised when the promoter 8-Br-cAMP was added.Conclusion PEMFs at 15 Hz 1 mT can promote type I collagen expression of bone marrow mesenchymal stem cells.and the effect is correlated with the cAMP-PKA pathway.

Objective To investigate the effects of an electromagnetic field on the extra-cellularly regulated kinase(ERK)signalling pathway and to determine the impact of electromagnetic activation on osteogenic proliferation and differentiation in rat bone marrow mesenchymal stem cells.Methods Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro.The third-passage cells were divided into 4 groups(Control,PD98059,EMF and EMF+PD98059).Western blotting Was used to detect the activation of the ERK signal pathway after exposure to an electromagnetic field.MTT assay Was used to determine the activation of proliferation in the celb in the different groups.The cells' alkaline phosphatase activities were also detected. Results (1)The ERK signal pathway in these rat bone marrow mesenchymal stem cells was activated after exposure to a 15 Hz.1 mT,sine wave form electromagnetic field for 5 min.Activation remained high for at least 1 h.PD98059 can effectively block the activation of the ERK signal pathway.(2)Cell proliferation was promoted after exposure to the electromagnetic field,and this effect could be significantly inhibited by PD98059.(3)Alkaline phosphatase was significantly elevated in these bone marrow mesenchymal stem cells after exposure to the electromagnetic field.The activation in the EMF+PD98059 group Was slightly greater than in the EMF group.Conclusion Electromagnetic fields of 15 Hz and 1 mT can activate the ERK signal pathway and alter proliferation and osteogenic differentiation in the bone marrow mesenchymal stem cells of rats.

Objective To screen the differential expression genes of bone marrow MSCs stimulated by electromagnetic field(EMF)with osteogenesis microarray analysis,and to study the underlying mechanism that EMF promotes the differentiation of bone marrow MSCs. Methods The Sprague-Dawley rat bone marrow MSCs were isolated and cultured in vitro.The third-passage cells who were stimulated by EMF and served as the stimulated group,and those who were not stimulated by the EMF served as the controls.Total RNA was extracted and purified,then it was used to synthesize cDNA and cRNA.The eRNA of stimulated group and the control group was hybridized with the rat oligo osteogenesis microarray,respectively.The hybridization signals were acquired by using X-ray film after chemiluminescent detection and the obtained data were analyzed using the web-based completely integrated GEArray Expression Analysis Suite.RT-PCR was used to identify the chosen genes BMP1,VDR and EGF. Results Nineteen differential expression genes were found between the stimulated group and the control group.There were 6 upregulated and 13 downregulated genes in the stimulated group.Semi-quantitative RT-PCR confirmed that the expression levels of BMP1,VDR mRNA in the stimulated group were upregulated and EGF downregulated. Conclusion The gene expression profiles about osteogenesis of the bone marrow MSCs were changed after EMF intervention(1 5 Hz,1 mT).These genes are involved in the difierentiation of bone marrow MSCs into osteoblast.These results provide deeper insight into the mechanism that EMF exposure facilitates the in vitro differentiation of bone marrow MSCs.