OBJECTIVE:To optimize the water extraction technology of Yinju jiedu oral liquid,and provide reference for the industrial production of the preparation. METHODS:According to the investigation of extraction time-extraction rate curves of chlo-rohenic acid of Yinju jiedu formula and extraction rate of chlorohenic acid in Lonicera japonica and other combined medicinal mate-rials in the formula,decoction methods and time of L. japonica were determined. Using the comprehensive scores of linarin,harpa-goside,(R,S)-epigoitrin,psoralen+angelicin contents and dry extraction yield as indexes,L9(34)orthogonal test was designed to detect the effects of adding water amount,decoction time times and optimize the extraction technology of the residues and other me-dicinal materials. Verification test was conducted. RESULTS:The optimal technology was L. japonica decocted first for 30 min with 8-fold water;the residues and other medicinal materials were decocted with 8-fold water for 3 times,1 h each time;combin-ing all the syrups. In verification test,the average contents of chlorohenic acid,linarin,harpagoside,(R,S)-epigoitrin,psoralen+angelicin were respectively 34.51,10.31,1.97,0.21,9.79 mg/g(RSD=1.24％,1.19％,1.40％,1.71％,1.28％,n=3);aver-age dry extraction yield was 25.4％(RSD=1.64％,n=3);average extraction rate of chlorohenic acid was 78.95％(RSD=1.24％,n=3). CONCLUSIONS:In the optimized water extraction technology,both the extraction rate of chlorohenic acid and contents of other ingredients are relatively high. The technology is stable and feasible.

Objective To study the protective effect of warming kidney and subsiding yang traditional Chinese medicine (TCM) method on lung tissues of rats with acute respiratory distress syndrome (ARDS) associated with sepsis. Methods Eighty healthy male Wistar rats were divided into normal control group, model group and low, medium and high dose Fusuheji groups by random number table, 16 rats in each group. The acute lung injury (ALI) model was established by injecting lipopolysaccharide (LPS) 3 mg/kg into a rat caudal vein within 5 minutes, and the normal control group was given the same volume of normal saline. Then the low, medium and high dose TCM groups were given low, medium and high dose Fusuheji TCM mixture (the ingredients of the mixture: radix aconite lateralis preparata 30 g, oysters 30 g, ginger 15 g, ephedra 15 g, licorice 10 g) 2.625, 7.875, 10.500 g/kg intragastric administration respectively. Equal volume of saline was given to the normal control group and model group by gavage. At 24 and 48 hours after respective administration, 8 rats were taken from each group to observe the pathological changes of lung tissues and score the lung injury. The rates of survival of rats were calculated after the experiment in various groups. Results After administration for 48 hours, the survival rate of rats in model group was obviously lower than that of the normal control group [18.7% (3/16) vs. 100.0% (16/16)], the low, middle and high dose Fusuheji groups' survival rates were all significantly higher than the rate of ALI model group [50.0% (8/16), 75.0% (12/16), 93.7%(15/16) vs. 18.7% (3/16), all P < 0.05]. There were no pathological changes in the lung tissues of rats in the normal control group, large amounts of exudates and hemorrhages were present in the lung tissues of ALI model group, and the inflammatory, exudative and hemorrhagic changes of lung tissues in the high, middle and low dose Fusuheji groups were obviously improved. After administration for 24 hours and 48 hours, the lung injury scores in the ALI model group were higher than those in control group (after administration for 24 hours: 7.83±0.60 vs. 2.89±4.23; after administration for 48 hours: 7.33±0.88 vs. 3.00±0.28), the scores of lung injury of any Fusuheji drug group were significantly lower than those of ALI model group, and the degrees of decrease were more marked in high dose Fusuheji group than those in low and middle dose Fusuheji groups (after administration for 24 hours: 3.37±0.32 vs. 6.00±0.44, 4.63±0.50; after administration for 48 hours: 3.25±0.25 vs. 5.25±0.25, 3.50±0.50). Conclusion The warming kidney and subsiding yang TCM method can improve the lung tissue injury in ARDS associated with sepsis in rats, promote the damaged lung tissue to recover, and ultimately the prognosis of ARDS rats is getting better.

Objective To screen the critical genes related to the development of esophageal squamous cell carcinoma ( ESCC ) by weighted gene co-expression network analysis ( WGCNA ) and to verify by experiments.Methods Gene expression data of ESCC were downloaded from gene expression omnibus (GEO) database based on gene chip platform ( GPL) 570, GPL571, GPL96/97 or GPL14613 platform, respectively. Meanwhile, the obtained differentially expressed genes together with gene expression data of 81 ESCC patients from the cancer genome atlas ( TCGA ) and clinical data were analyzed by WGCNA to set up co-expression networks including mRNA and microRNA ( miRNA ) . The expression of miRNA in ESCC tissues and paracancerous tissues was examined by quantitative real-time polymerase chain reaction ( RT-PCR ) .And the expression of target protein Kruppel like factor 4 ( KLF4 ) and desmocollin 2 ( DSC2 ) were detected by immunohistochemistry .After ESCC cell line ECA-109 cells were transfected with miRNA-92b-3p mimic, cell cycle was tested by flow cytometry ,the cell invasion and migration ability was measured by Transwell chamber assay and scratch-wound assay.The expression of KLF4 and DSC2 was observed by confocal laser scanning microscopy and Western blotting .The target genes were verified by luciferase assay .T-test, rank sum test, chi-square test and Pearson correlation analysis were performed for statistical analysis .Results A total of 4023 differential expression gene ( DEG) and 328 differential expression miRNA ( DEM) were screened and 11 gene modules were set up by WGCNA .Among them, the brown modules were negatively associated with tumor grade and T stage (r=-0.340 and -0.268, P=0.002 and 0.016).Meanwhile, has-miR-92b and the potential target genes KLF4 and DSC2 were all in the brown module .Furthermore, the results of RT-PCR showed the expression of miRNA-92b-3p in ESCC tissues was higher than that in paracancerous tissues (3.052(1.652, 5.371) vs.0.985(0.558, 2.032)), and the difference was statistically significant (Z=-4.021,P<0.01). The results of immunohistochemistry demonstrated that the positive rates of KLF 4 and DSC2 in ESCC tissues were 43.3％(13/30) and 20.0％(6/30), respectively, which were lower than those of paracancerous tissues (70.0％(21/30) and 63.3％(19/30)), and the differences were statistically significant (χ2 =4.344 and 1.589, both P<0.05).After ECA-109 cells were transfected with miRNA-92b-3p mimics, the percentage of cells at G0/G1 phase decreased ((63.71 ±2.83)％vs.(54.62 ±4.00)％) and the percentage of cells at the S phase and G2/M phase increased ((31.81 ±2.88)％vs.(41.20％±2.87)％, and (3.87 ±1.75)％vs. (8.10 ±1.71)％, respectively), and the differences were statistically significant (t =3.215, 4.000 and 2.998;P=0.032, 0.016 and 0.040).The invasion and migration ability of the cells were significantly improved (79.67 ±27.54 vs.280.33 ±46.18, (69.72 ±3.91)％ vs.(84.90 ±5.25)％), and the differences were statistically significant (t=6.465 and 4.019, P=0.003 and 0.016).The results of Western blotting indicated that, compared with control mimic group , the expression of KLF4 and DSC2 was both dramatically downregulated after transfected with miRNA-92b-3p mimics transfected (1.00 ±0.23 vs.0.42 ±0.03, 1.00 ±0.20 vs.0.55 ± 0.21), and differences were statistically significant (t=4.470 and 5.493, P=0.042 and 0.032).The results of luciferase assay demonstrated that miRNA-92b-3p could directly bind KLF4 and DSC2. Conclusion WGCNA is an efficient systemic biological approach by which miRNA-92b-3p is identified as a new cancer-promoting gene .