Objective To investigate the relationship between bed rest time and development of prethrombotic state in the elderly patients with hip fracture.Methods One hundred and sixty-six patients who stayed in bed after hip fracture,aged ≥65 yr,were divided into 5 groups according to the bed rest time on admission to hospital:bed rest time ＜ 24 h (Ⅰ group,n =61),bed rest time 24-48 h (Ⅱ group,n =29),bed rest time 3-6days (Ⅲ group,n =29),bed rest time 7-14 days (Ⅳ group,n =34),and bed rest time ＞ 14 days (Ⅴ group,n =13).Venous blood samples were collected to measure the platelet count,coagulation function,and concentrations of plasma D-Dimer and serum α-granule membrane protein 140 (GMP-140).The development of abnormality in each index was recorded.The development of deep vein thrombosis in both lower extremities was diagnosed using color Doppler ultrasound in D-Dimer-positive patients.Results Compared with group Ⅰ,the abnormal rate of fibrinogen (Fib) and D-Dimer and serum GMP-140 concentrations were significantly increased in Ⅱ and Ⅲ groups,the abnormal rate of platelet count,Fib and D-Dimer and serum GMP-140 concentrations were increased in lⅣ group,and the abnormal rate of platelet count was increased,and no significant change was found in the serum GMP-140 concentrations and abnormal rate of Fib and D-Dimer in Ⅴ group.Compared with group Ⅱ,the serum GMP-140 concentrations were significantly increased in Ⅲ and Ⅳ groups,the abnormal rate of Fib and D-Dimer was increased in Ⅳ group,and no significant change was found in the abnormal rate of Fib and DDimer in Ⅲ group.The abnormal rate of platelet count was significantly lower in Ⅳ group than in Ⅲ group.Color Doppler Ultrasonography showed no sign of deep vein thrombosis.Conclusion For the elderly patients with hip fracture,the possibility of pre-thrombotic state developed is increased when the bed rest time is more than 24 h,and the patients were classified as high-risk patients when the bed rest time is more than 3 days.

AIM: To evaluate the effect of GYY4137, a novel hydrogen sulfide (H2S) donor, on cytosolic lipid decomposition in mouse primary steatosis hepatocytes.METHODS: Oleic acid (OA) was used to induce hepatic steatosis model in vitro.The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion were divided into 4 groups: the cells in control group were incubated with normal medium for 54 h;the cells in model group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 for 6 h;the cells in H2S group or DL-propar-gylglycine (PAG;an inhibitor of cystathione γ-lysase, inhibiting H2S synthesis) group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 which contained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h.The glycerin release and the protein expression of hormone-sensitive lipase (HSL) in the cells were mea-sured.RESULTS: Compared with model group, the glycerin release and the protein expression of phosphorylated HSL (p-HSL) in H2S group decreased significantly, while those increased significantly in PAG group.CONCLUSION: In steatosis hepatocytes, exogenous H2S possibly decreases cytosolic lipid decomposition by decreasing the protein level of p-HSL.

Objective Increasing evidence suggest that aberrant activation of PI3K/Akt is involved in many human cancers, and that inhibition of the PI3K/Akt pathway might be a promising strategy for cancer therapy. The study is to evaluate the effects of combined therapy of PI3K inhibitor (LY294002) and Ad-PTEN in athymic mice xenogeneic transplant model of human glioma and to reveal the possible mechanisms involved.Methods Twenty-four athymic mice were randomly divided into 4 groups (DMSO、Ad-vector plus DMSO、LY294002 alone and Ad-PTEN plus LY294002), and were treated, respectively. Athymic mice xenogeneic transplant model was established by inoculation (sc) with LN229 glioma cells. Body mass (BM) and diameter of tumor mass were measured. Furthermore, The protein expressions of PTEN、p-Akt、CyclinD1、Caspase-3、MMP-2、p-FAK in tumor tissues were analyzed with immunohistochemistry.Results The tumor-inhibiting rate of was significantly higher in Ad-PTEN plus LY294002 than in the LY294002 alone (92.46 vs 65.59%)( P <0.05).The protein expressions of PTEN and Caspase-3 were significantly higher, while PCNA、CyclinD1、bcl-2 and MMP-2、p-FAK was significantly lower in Ad-PTEN plus LY294002 group than in the other three groups ( P <0.05).Conclusions LY294002 plus Ad-PTEN achieve better outcome than either alone in treating glioma possibly through enhancement of the inhibitory action of PI3K/Akt pathway and Ad-PTEN pathway.

ObjectiveTo investigate the mechanism of action of Sini powder in the treatment of liver cancer based on network pharmacology and molecular docking. MethodsTraditional Chinese Medicine Systems Pharmacology Database and Analysis Platform was used to obtain the compound and target of Sini powder, and the corresponding gene Symbol was obtained through Uniprot. The disease genes of liver cancer were obtained from Human Genome Database, and the genes with intersection with the target genes of Sini powder were screened out. Cytoscape3.7.1 software was used to draw the map of “traditional Chinese medicine (TCM)-compound-target” network. STRING was used to construct a protein-protein interaction (PPI) network, R studio software was used to conduct gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses on therapeutic targets, and then the results were visualized. The active component with the highest number of targets was selected as the ligand, and the target with the highest degree in the PPI network was selected as the receptor, so as to predict the structure of receptor-ligand complex and the amino acid residues that bind to each other. ResultsIn this study, 91 core targets and 141 relevant active components of Sini powder were screened out, among which quercetin and kaempferol were the main active components in the treatment of liver cancer. TP53 and HSP90AA1 were the main therapeutic targets. The GO enrichment analysis obtained 1007 items which met the screening criteria, which were mainly involved in the biological processes of antioxidation reaction, activity regulation of protein serine and threonine kinase, and cellular stress response. The KEGG enrichment analysis obtained 102 pathways, which mainly regulated the hepatitis B pathway and the PI3K-Akt signaling pathway in the prevention and treatment of liver cancer. The results of molecular docking showed a synergistic antitumor effect between the crystal structure domains VAL147, CYS220, GLU221, and PRO222 of quercetin-TP53. ConclusionThis study reveals the mechanism of action of Sini powder in the treatment of liver cancer by acting on multiple targets and signaling pathways, which provides a theoretical basis for biological experiments.

Objective To evaluate the value of MRI texture analysis based on gray level co-occurrence matrix to predict cervical lymph node metastasis in patients with tongue carcinoma. Methods A total of 70 patients with tongue carcinoma confirmed by pathology were analyzed retrospectively. The patients were divided into cervical lymph node (LN) metastasis group (unilateral LN+, n=18;bilateral LN+,n=22) and no cervical lymph node metastasis(LN-,n=30). T1W, T2W and contrast-enhanced T1W images of the largest section of tumor were selected. ROI of the lesion was manually drew and GLCM texture parameters (energy, contrast, correlation, inverse difference and entropy) were extracted. The tumor length, thickness and para-lingual distance between tumor and tongue midline were also measured.Differences of all parameters were compared between LN+ group and LN- group, unilateral and bilateral cervical lymph node metastasis group, the parameters with statistically significant difference in predicting the efficiency of cervical lymph node metastasis were analyzed. The diagnostic efficiency of lymph node metastasis was calculated. Results The correlation, inverse difference and entropy based on T2WI showed significant difference (Zcor elation=2.97, tinverse dif erence=5.14, tentropy=2.41; P<0.05), entropy showed the best diagnostic efficiency, the area under the ROC curve (AUC) was 0.90, the cut off value was 7.19, the sensitivity was 80.0％, specificity was 86.7％. Only entropy showed significant difference between unilateral LN+group and bilateral LN+group (P<0.05), the AUC was 0.82, the cut off value was 7.47, the sensitivity was 90.9％, specificity was 78.8％. The index of tumor length, thickness and para-lingual distance between tumor and tongue midline all showed significant difference between LN+group and LN-group (P<0.05), the thickness showed the best diagnostic efficiency, the AUC value was 0.81, the cut off value was 11.19, the sensitivity was 78.0％, specificity was 81.7％. The index of tumor length, thickness and para-lingual distance between tumor and tongue midline showed no significant difference between unilateral LN+ group and bilateral LN+ group (P>0.05). The diagnostic sensitivity of radiologists was 65.0％ (26/40), the specificity was 80.0％ (24/32) on cervical lymph node metastasis. Conclusion Texture analysis based on T2WI can predict cervical lymph node metastasis in patients with tongue carcinoma. Entropy has certain value in predicting bilateral cervical lymph node metastasis.

Objective To investigate clinical characteristics and risk factors of different stages of urinary tract infection after kidney transplantation.Methods Clinical data of 209 kidney transplant recipients were retrospectively analyzed.According to time points of postoperative follow-up,all recipients were divided into 3 stages:within 1 month post-kidney transplantation,1-6 months post-kidney transplantation,and 7-12 months post-kidney transplantation.The incidence of urinary tract infection,urine culture results of recipients with urinary tract infection and drug resistance characteristics of common pathogens during different stages after kidney transplantation were analyzed.The strains of patients with recurrent urinary tract infection were identified.The risk factors of urinary tract infection and the effect of urinary tract infection on renal allograft function were analyzed.Results The urinary tract infection rate was 90.0％in the first stage,49.3％in the second stage and 22.5％in the third stage.The urinary tract infection rates of male recipients undergoing living-related organ donation in the second and third stages were lower than those of female recipients(both P＜0.05).Urine culture test yielded positive results in 60 cases,and 84 strains of pathogenic bacteria were detected,mainly Gram-negative bacteria,among which Klebsiella pneumoniae accounted for the highest proportion.Sixty-six recipients had recurrent urinary tract infection,and the detected pathogens included Klebsiella pneumoniae,Escherichia coli and Candida glabrata,etc.Univariate analysis showed that postoperative use of antithymocyte globulin was the risk factor for urinary tract infection in the first stage.Preoperative urinary tract infection and donor type were the risk factors for urinary tract infection in the second stage.Gender and age of the recipients were the risk factors for urinary tract infection in the third stage.Multivariate analysis revealed that postoperative use of antithymocyte globulin was the risk factor for urinary tract infection in the first stage.Gender and age of the recipients were the risk factors for urinary tract infection in the third stage(all P＜0.05).In the third stage,65 cases were cured and 38 cases were not cured.In the treated recipients,the serum creatinine level and white blood cell count were decreased after corresponding treatment than those before treatment(both P＜0.05).Conclusions Gram-negative bacteria are the main pathogens of urinary tract infection in kidney transplant recipients,and drug resistance is relatively high.Postoperative use of antithymocyte globulin,female and old age are the risk factors for urinary tract infection in kidney transplant recipients.

OBJECTIVETo observe the effect difference between acupuncture of "five needles therapy" and conventional acupuncture for asthma of latent cold phlegm-fluid in the lung.

METHODSTwo hundred and ten cases were randomly assigned into an observation group and a control group, 105 cases in each one. Finally 7 cases were dropped out in the observation group; 6 cases in the control group. Feishu (BL 13), Dazhui (GV 14), Fengmen (BL 12) were used in the observation group; conventional acupuncture was used in the control group, and the main acupoints were Feishu (BL 13), Zhongfu (LU 1), Tiantu (CV 22), Danzhong (CV 17), Kongzui (LU 6), Dingchuan (EX-B 1), Fenglong (ST 40), Fengmen (BL 12), Taiyuan (LU 9). The needles were retained for 30 min each time, once a day for continuous 12 days. The scores of the individual symptoms and signs were observed before treatment and on the 3rd, 6th, 9th, 12th days, including pant, cough, cough up phlegm, fullness and oppression in the chest and diaphragm, wheezing rale and shortness of breath. The clinical effects were compared between the two groups.

RESULTSThe scores of six individual symptoms and signs on the 3rd, 6th, 9th, and 12th days in the two groups were lower than those before treatment (all<0.05), except the score of wheezing rale in the control group on the 3rd day (>0.05). The scores of pant, wheezing rale, cough on the 3rd, 6th, 9th, and 12th days in the observation group were lower than those in the control group (all<0.05), except the score of wheezing rale score on the 3rd day (>0.05). There were no significant difference between the two groups about the scores of cough up phlegm, fullness and oppression in the chest and diaphragm and shortness of breath on the 3rd, 6th, 9th, and 12th days (all>0.05), except the score of fullness and oppression in the chest and diaphragm in the observation group was lower than that in the control group on the 12th day (<0.05). 46 cases were clinical cured, 39 cases were markedly effective, 10 cases were effective and 3 cases were ineffective in the observation group with the total effective rate of 96.9%. 23 cases were clinical cured, 43 cases were markedly effective, 24 cases were effective and 9 cases were ineffective in the control group with the total effective rate of 90.9%. The difference was statistical (<0.05).

CONCLUSION"Five needles therapy" has significant therapeutic effect for asthma of latent cold phlegm-fluid in the lung, which is better than conventional acupuncture.

ObjectiveTo explore the mechanisms of internal treatment （Renshen Baidusan）， external treatment （Yurui Enema）， and combination of the two methods in treating intestinal mucosal injury in the rat model of ulcerative colitis （UC） from the changes of phosphatidylinositol-3 kinase （PI3K）/protein kinase B （Akt）/nuclear factor-κB （NF-κB） pathway. MethodFifty SPF-grade SD rats were randomized into blank， model， Renshen Baidusan （15.6 g·kg^-1）， Yurui Enema （25 g·kg^-1）， and combined treatment （15.6 g·kg^-1 Renshen Baidusan + 25 g·kg^-1 Yurui Enema） groups （n=10）. The rat model of UC was established in other groups except the blank group by 2，4， 6-trinitrosulfonic acid （TNBS）/ethanol. The rats were administered with corresponding drugs once a day for 14 consecutive days since the 8th day after modeling. The histopathological changes of colon were observed by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of tumor necrosis factor （TNF）-α， interferon （IFN）-γ， interleukin （IL）-4， and IL-10 in the colon tissue. The apoptosis of colon epithelial cells was detected by terminal deoxynucleotidyl transferase-mediated nick end labeling （TUNEL）. The location and expression of B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， TNF-α， and IL-6 in the colon tissue were examined by immunohistochemistry. Real-time quantitative fluorescence polymerase chain reaction （Real-time PCR） and Western blot were employed to determine the mRNA and protein levels， respectively， of the proteins in the PI3K/Akt/NF-κB pathway in the colon tissue. ResultIn the model group， HE staining showed a large number of inflammatory cell infiltration in the mucosa and submucosa. Compared with the blank group， the model group showed elevated levels of TNF-α and IFN-γ and lowered levels of IL-4 and IL-10 in the colon tissue， increased apoptosis rate of colon epithelial cells， increased positive expression of Bax， TNF-α， and IL-6， and decreased positive expression of Bcl-2 （P<0.05）. Moreover， the model group showed up-regulated mRNA levels of PI3K， Akt， and NF-κB and protein levels of PI3K， p-PI3K， Akt， p-Akt， p65， p-p65， Bax， and cleaved Caspase-3， increased Bax/Bcl-2 and cleaved Caspase-3/Caspase-3 ratios， and down-regulated protein levels of NF-κB suppressor protein α（IκBα）， Bcl-2， and Caspase-3 in the colon tissue （P<0.05）. Compared with the model group， the internal treatment， the external treatment， and the combination （referred to as the three groups） alleviated the colonic mucosal injury， lowered the levels of TNF-α and IFN-γ and elevated the levels of IL-4 and IL-10 in the colon tissue， decreased the apoptosis rate of colon cells， inhibited the positive expression of Bax， TNF-α， and IL-6， and promoted the positive expression of Bcl-2 （P<0.05）. Furthermore， the combination group down-regulated the mRNA level of PI3K （P<0.05）. The three groups down-regulated the mRNA levels of Akt and NF-κB and the protein levels of p-PI3K， Akt， p-Akt， p65， p-p65， Bax， and cleaved Caspase-3 in the colon tissue， decreased the Bax/Bcl-2 and cleaved Caspase-3/Caspase-3 ratios， and up-regulated the protein levels of IκBα， Bcl-2， and Caspase-3 （P<0.05）. ConclusionRenshen Baidusan， Yurui Enema， and their combination may inhibit the activation of PI3K/Akt/NF-κB signaling pathway and regulate the expression of genes and proteins related to this pathway to achieve anti-inflammatory and anti-apoptotic effects， thus restoring the intestinal mucosal barrier function of UC rats.

ObjectiveTo study the mechanism of Renshen Baidusan in regulating adenylate-activated protein kinase （AMPK）/Unc-51-like kinase 1 （ULK1） autophagy pathway to inhibit mucosal barrier damage in the mouse model of ulcerative colitis （UC）. MethodSixty SD rats were randomized into normal， model， sulfasalazine enteric-coated tablets （0.312 5 g·kg^-1， western medicine）， and high-， medium-， and low-dose （31.2， 15.6， 7.8 g·kg^-1， respectively） Renshen Baidusan groups. The UC model was induced by 2，4，6-trinitrobenzenesulfonic acid （TNBS）/50% ethanol. The drugs were administrated by gavage for 2 weeks， and then the histopathological changes of the colon were examined. Real-time quantitative polymerase chain reaction was conducted to measure the mRNA level of AMP-activated protein kinase subunit alpha （AMPKα）. Western blot was employed to determine the protein levels of closure protein （Occludin）， compact linking protein-2 （Claudin-2）， autophagy marker p62， microtubule-associated protein 1 light chain 3B （LC3B）， phosphorylated AMPK （p-AMPK）， and phosphorylated ULK1 （p-ULK1）. ResultCompared with the normal group， the model group showed increased colon injury score （P<0.05）， down-regulated mRNA level of AMPKα （P<0.05） and protein levels of p-AMPK， p-ULK1， and Occludin， decreased LC3Ⅱ/Ⅰ ratio （P<0.05）， and up-regulated protein levels of p62 and Claudin-2 （P<0.05）. Compared with the model group， all the doses of Renshen Baidusan lowered the colon injury score， up-regulated the mRNA level of AMPKα and the protein levels of p-AMPK， p-ULK1， and Occluding， increased LC3Ⅱ/Ⅰ ratio， and down-regulated the protein levels of p62 and Claudin-2. Moreover， the medium-dose group showed a significant intervention effect （P<0.05）. ConclusionRenshen Baidusan can protect the intestinal mucosal barrier from damage， and the medium dose showed the best efficacy. It may activate the AMPK/ULK1 pathway to accelerate the transformation of LC3Ⅰ to LC3Ⅱ and promote the degradation of p62， so as to improve the function of Occludin and Claudin-2 and repair the mechanical damage of the intestinal barrier.