Objective To explore the curative effect of the traditional Chinese medicine"SHU JI FANG"in treating lumbar discogenic low back Pain(DLBP).Methods 156 cases were divided into three groups by random with 52 cases in each group.A treatment group was treated by traditional Chinese medicine"SHU JI FANG".A control group 1 was treated by Bextra;A control group 2 was treated bv Chinese traditional medicine Zuogui pill.Results The difiererlce of cure rate and the excellent rate between the treatment group and the control group I did not show significance by X2 check,with P＞0.05;while the difference between the treatment Group and the Control group 2 showed significance by X2 check with P＜0.05.Conclusion I"SHU JI FANG"is effective in treating discogenic low back pain.

Objective To study the methylation status of secreted frizzled-related protein (SFRP) 1 and SFRP2 genes in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) and the relationship between the methylation status of the two genes and the development of HCC.Methods Using methylation-specific polymerase chain reaction (MSP) to detect methylation status of SFRP1 and SFRP2 genes of 45 specimens of HCC tissue and adjacent non-tumorous liver tissue from HCC patients during operations,and 6 normal liver tissues from patients with cholecystolithiasis or hepatic hemangiomas. The data were analyzed by chi-square test and Fisher exact test. Results SFRP1 gene methylation was detected in 28 HCC tissues and 16 adjacent non-tumorous liver tissues,accounted for 62.2% and 35.6%,respectively;and SFRP2 gene methylation was detected in 23 HCC tissues and 13 adjacent non-tumorous liver tissues,accounted for 51.1% and 28.9%,respectively;while no methylation was detected in 6 samples of normal liver tissues. There was no significant difference between the methylation of SFRP1 and SFRP2 genes in HCC tissues and gender,age,HBV serum markers,types of adjacent non-tumorous liver tissues,metastasis and pathological stage (P＞0.05).The abnormal methylation status between SFRP1 and SFRP2 genes was linear correlated in HCC tissues (r=0.381,P=0.01).Conclusion Hypermethylation of SFRP1 and SFRP2 genes frequently occurs in HBV-related HCC,which may be an important molecular biomarker for prediction of hepatocarcinogenesis in the future.

Objective To study the effects of Zhishang Sanfang (ZS) in promoting the union of radius fracture. Methods Twenty-eight rabbit models with fr acture were randomly allocated to ZS group (Group A) and control group (Group B).His tological, histochemical and histomorphological quantitative detection was used to observe the morphological features of the frature end in the 14th and 28th da y after operation. Results The formation of blood capillaries and bone trab eculae in Group A was superior to those in Group B. The scores of the four callu s and th e depth of the lateral callus in Group A were higher than those in Group B. Con clusion ZS has obvious effect in promoting the union of fracture.

Objective To explore the effect of Botulinum toxin type A (BTX-A) injection combined with rehabilitation training and medicated bath on spasm for children with cerebral palsy. Methods 80 children with spastic and mixed cerebral palsy were randomly divided into two groups with 40 cases in each group. The control group received physical therapy, and the observation group received BTX-A injection and rehabilitation training and medicated bath. They were assessed with modified Ashworth scale (MAS) and Gross Motor Function Measure (GMFM-88) before and 3 months after treatment. Results The scores of MAS and GMFM were better in 2 groups after treatment (P<0.05) and the observation group was better than that of the control group (P<0.05). Conclusion BTX-A injection combined with medicated bath can reduce muscle tension, improve gross motor function of children with spastic and mixed cerebral palsy.

Objective:To explore the application of pyrophosphorolysis-activated polymerization(PAP)to monitor plasma cfDNA in ad-vanced non-small cell lung cancer(NSCLC).Methods:A total of 85 patients diagnosed with advanced NSCLC between March 2016 and June 2017 were enrolled in the present study. EGFR mutations in cfDNA extracted from the plasma were detected using PAP and ARMS-PCR technology.The concordance analysis of EGFR mutations involved plasma vs.tumor tissue and PAP vs.ARMS-PCR.Further-more,38 EGFR-positive patients were selected to monitor EGFR mutations with PAP.Results:No statistical differences in EGFR muta-tions were observed between plasma and tumor tissue(P=0.092),as well as PAP and ARMS-PCR(P=0.210).The detection rate of EGFR mutations in cfDNA was higher in the progressor than in the non-progressor(62.5% vs.21.3%,P<0.001).Conclusions:PAP can be used for detecting and monitoring EGFR mutations in cfDNA to predict disease progression.

Objective:To investigate the value of pericoronary adipose tissue histogram parameters based on coronary CT angiography (CTA) images for the differentiation of acute coronary syndrome and stable coronary artery disease.Methods:The clinical data and CTA images of 93 patients with coronary CTA examination in Suzhou Kowloon Hospital from 2013 to 2018 were analyzed retrospectively. There were 39 patients with acute coronary syndrome (acute coronary syndrome group) and 54 patients with stable coronary artery disease (stable coronary artery disease group). A region of interest (ROI) was drawn around the stenosis of the coronary arteries, with CT attenuation ranging from-190 to -30 HU to exclude non-adipose tissue. The CT attenuation of ROI excluding non-adipose were measured and histogram analysis was performed. The obtained parameters included the mean value, median value and the 5th, 10th, 45th, 55th, 70th and 95th percentiles. The differences in histogram parameters between the two groups were compared, and then the value of each parameter in differentiating acute coronary syndrome and stable coronary artery disease was evaluated based on receiver operating characteristic (ROC) analysis. The stepwise regression of multivariate logistic regression analysis was used to identify the useful features and establish the final prediction model. The ROC curve of the final model was calculated and its value was analyzed.Results:The mean, median and the 5th, 10th, 45th, 55th,70th and 95th percentile differences between the acute coronary syndrome group and the stable coronary artery disease group were statistically significant (all P<0.05). The ROC curve for the median and the 95th percentile had the same area under curve (AUC) of 0.73. The sensitivity, specificity and AUC of the diagnostic model established by multiple logistic regression were 82.1%, 89.1% and 0.90 respectively. Conclusion:CT attenuation histogram of pericoronary adipose tissue is of high value in differentiating acute coronary syndrome from stable coronary artery disease.

BACKGROUND:Cartilage degeneration and subchondral bone damage are the main pathological features of osteoarthritis,and treatment based on this pathological feature will be a promising improvement for osteoarthritis. OBJECTIVE:To design and study an annotated strontium ranelate-loaded drug delivery system and to observe its therapeutic effect on promoting cartilage repair and improving subchondral bone structure in osteoarthritis. METHODS:(1)In vitro experiment:Strontium ranelate was loaded into sodium alginate/collagen hydrogel matrix to construct in situ drug delivery system,and the in vitro slow release performance of the system was characterized.Strontium ranelate-loaded sodium alginate/collagen hydrogel(experimental group)and alginate sodium/collagen hydrogel(control group)were co-cultured with bone marrow mesenchymal stem cells,respectively,and cultured cells were used as a blank control group to detect cell proliferative activity.After chondroblast-induced differentiation,saffron O staining,Alcian blue staining and RT-qPCR were performed respectively.The two hydrogels were co-cultured with osteoblasts,and the cultured cells were used as a blank control group for immunofluorescence staining and RT-qPCR.(2)In vivo experiment:A total of 18 adult SD rats were selected and the model of right posterior knee osteoarthritis was established by the method of medial meniscectomy.After 1 week,the rats were divided into three groups by the random number table method:The blank group did not receive any treatment.The control group was injected with sodium alginate/collagen hydrogel in the knee,and the experimental group was injected with strontium ranelate-loaded sodium alginate/collagen hydrogel,with 6 rats in each group.After 6 weeks,the samples were subjected to Micro-CT scanning,hematoxylin-eosin staining,saffron O-solid green staining and immunofluorescence staining. RESULTS AND CONCLUSION:(1)In vitro experiment:Strontium ranelate-loaded sodium alginate/collagen hydrogel had porous microstructure and sustainable release of strontium ranelate.At 21 days,the cumulative release reached(60.89±0.58)%.Bone marrow mesenchymal stem cell staining showed that both hydrogels had good cytocompatibility.The results of the CCK-8 assay demonstrated that strontium ranelate-loaded sodium alginate/collagen hydrogel could promote the proliferation of bone marrow mesenchymal stem cells.The results of Safranin O staining,Alcian blue staining,immunofluorescence staining and RT-qPCR exhibited that strontium ranelate-loaded sodium alginate/collagen hydrogel could promote chondrogenic differentiation of bone marrow mesenchymal stem cells.Immunofluorescence staining and RT-qPCR revealed that strontium ranelate-loaded sodium alginate/collagen hydrogel could decrease bone resorptivity by increasing the ratio of osteophosphorin/nuclear factor κB receptor activator ligand.(2)In vivo experiment:Micro-CT scan verified that compared with the blank group and control group,the subchondral bone volume fraction and bone mineral density of the knee of rats were increased in the experimental group(P<0.05,P<0.01).Histological staining displayed that compared with the blank group and control group,the knee cartilage injury was significantly reduced;the expression of type II collagen was promoted,and the expression of matrix metalloproteinase 2 protein was inhibited in the experimental group(P<0.05,P<0.01).(3)These results confirm that the strontium ranelate-loaded sodium alginate/collagen hydrogel can promote the repair of cartilage defects in osteoarthritis and reconstruct the complex interface between cartilage and subchondral bone.

BACKGROUND:Stem cell transplantation is a new way to prevent and cure intervertebral disc degeneration.However,whether the transplanted stem cells can survive,proliferate,differentiate,and restore the function of nucleus pulposus cells after transplantation,is the key and difficult point to overcome. OBJECTIVE:To explore the effects of Bushenhuoxue decoction on survival,proliferation,and nucleus pulposus-like differentiation of adipose-derived stem cells. METHODS:A Transwell chamber was used to construct a co-culture model of human adipose-derived stem cells and human degenerative nucleus pulposus cells.The experiment was divided into control group,model group,drug-containing serum group,and drug-free serum group.Except for the control group,the co-culture system of other groups was treated with 50 μmol/L tert-butyl hydrogen peroxide for 24 hours.The drug-containing serum group and drug-free serum group were treated with DMEM low-glucose complete culture medium containing drug-containing serum of Bushenhuoxue decoction or drug-free serum with 20％volume fraction for 48 hours.The sublayer adipose-derived stem cells were taken.Toluidine blue staining was used to detect proteoglycan synthesis levels.Real-time PCR method was used to detect mRNA expression of type Ⅱ collagen,proteoglycan and SRY-box transcription factor 9.The protein expression of SOX9 was detected by western blot assay.Lactate dehydrogenase assay was used to detect cytotoxicity.Flow cytometry was used to detect reactive oxygen species,and β-galactosidase staining was used to detect cell senescence. RESULTS AND CONCLUSION:(1)Compared with the control group,the proportion of necrotic cells in the model group increased;toluidine blue staining became lighter,and the expression levels of type Ⅱ collagen,proteoglycan,SOX9 mRNA and SOX9 protein decreased(P<0.05).Compared with the model group,the drug-containing serum of Bushenhuoxue decoction could significantly reduce cell injury and promote the expression of type Ⅱ collagen,proteoglycan,SOX9 mRNA,and SOX9 protein(P<0.05),but the improvement in the drug-free serum group was not significant(P>0.05).(2)Compared with the control group,the contents of cytotoxicity,reactive oxygen species,and cell senescence in the model group were significantly increased.Compared with the model group,the microenvironment of the coculture system was significantly improved by drug-containing serum of Bushenhuoxue decoction(P<0.05),while drug-free serum had no significant effect on the microenvironment of the co-culture system(P>0.05).(3)The results show that Bushenhuoxue decoction can promote the survival,proliferation,and nucleus pulposus-like differentiation of adipose-derived stem cells.

OBJECTIVE：To investigate the correlation of storage life and effective composition content with color value of Carthamus tinctorius ，and to provide reference for the quality evaluation of C. tinctorius with different years of storage. METHODS：Using 24 batches of C. tinctorius from same place of production with different years of storage （0，1，2 years，8 batches each type ）as samples ，the contents of hydroxysafflor yellow A （HSYA）and kaempferol were determined by HPLC. Color value [lightness value （L*），red-green value （a*），yellow-blue value （b*）] were determined by spectrophotometer. SPSS 19.0 statistical software was used to analyze the correlation of storage life and effective composition content with color value. RESULTS：Kaempferol content was still high after 1 year or 2 years of storage （0.161％，0.061％，respectively）. However ，the content of HSYA decreased with the prolongation of the storage life （the average content of HSYA were 2.46％，1.58％，and 1.51％ after storage 0，1 and 2 years，respectively），and the color of the drug became darker （a* value decreased ）. Results of correlation analysis showed that the content of HSYA was positively associated with color value L*，a*（r＝0.430，0.781，P＜0.05 or P＜0.01）；the content of HSAY was negatively associated with storage life （r＝－0.777，P＜0.01）. There was no correlation between the remaining variables （P＞0.05）. CONCLUSIONS ：The longer the storage life ，the darker the color and the lower the content of HSYA ，so it is not suitable for over year and multiyear preservation.

OBJECTIVE：To investigate the anti-inflammatory activity of 70％ ethanol extracts from Garcinia oblongifolia （GOEE）on LPS-induced RAW 264.7 cells and its potential molecular mechanism. METHODS ：GOEE was obtained after the fresh G. oblongifolia epicarp refluxed with 70％ ethanol. The contents of total phenol and total flavonoids were determined by Folin-Ciocalteau assay and UV spectrophotometer. MTT assay was used to detect the cytotoxicity of different doses of GOEE. The inflammatory model was induced in RAW 264.7 cells by lipopolysa- ccharide （LPS）. Using dexamethasone and N-acetyl-L-cysteine as positive control ，Griess assay and 2′，7′-dichloro- fluorescein assay were used to detect the contents of NO in cell culture medium and ROS in cells. The levels of TNF-α，IL-6，and IL- 1β in cell culture medium were measured by ELISA. The protein expression of p 65，p-p65，IκBα，p-IκBα，HO-1 in cells and NRF 2 in nucleus were determined by using Western blotting assay. RESULTS：The contents of total phenol and flavonoids in GOEE were （20.191±1.264）and（12.571±0.020）mg/g，respectively. At the concentration below 500 μ g/mL， GOEE had no significantly effect on survival rate of RAW 264.7 cells（P＞ 294043）0.05）. Compared with control group ，the contents of NO and ROS，the levels of TNF-α，IL-6 and IL- 1β，ratio of p-p 65 top65，ratio of p-IκBα to IκBα，protein expression of NRF 2 were increased significantly in LPS model group （P＜0.05 or P＜0.01）. Compared with LPS model group ，the contents of NO（except for GOEE 50 μg/mL group）and ROS ，the levels of TNF-α，IL-6 and IL- 1β，ratio of p-p 65 to p 65 and ratio of p-IκBα to IκBα were decreased significantly in GOEE groups and positive control groups ，while protein expression of HO- 1 and NRF 2 were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS：GOEE attenuates LPS-induced macrophages inflammation injury by inhibiting the inflammatory response and the phosphorylation of NF-κB pathway，promoting NRF 2 protein transportation to the nucleus.