Objective: To prepare highly purified THANK protein. Methods: THANK was efficiently expressed in E.coli as inclusion bodies. After bacteria were lysed under ultrasonication, TE buffer,1% TritonX and 2 mol /L urea was used to efficiently extract inclusion bodies. After denaturation with 8 mol/L urea,THANK was partially purified by Sephacryl S-200 chromatography, and then subsequent refolding step was optimized for a maximal recovery of biological active protein. The renatured THANK was purified to homogeneity with Q Sephadex Fast Flow gel filtration and then desalted by Sephadex G-25 chromatography. Results: The purity of biologically active THANK was above 97% in SDS-PAGE densitometric studies. Conclusion: An effective method of denaturation, renaturation and purification of recombinant THANK is established.

Objective To investigate the clinical significance of autophagy-related protein 7 (ATG7) in the diagnosis of HBV-related hepatocellular carcinoma (HBV-HCC) by measuring the expression level of serum ATG7 in patients with HBV-HCC. Methods A total of 50 patients with chronic hepatitis B (CHB) and 89 patients with HCC who were hospitalized in Mengchao Hepatobiliary Hospital of Fujian Medical University from June 2018 to December 2020 were enrolled, among whom 67 patients had HBV-HCC (HBV-HCC group) and 22 patients had no HBV-HCC (non-HBV-HCC group), and 20 healthy volunteers who underwent physical examination were enrolled as healthy control (HC) group. Demographic data and laboratory data including alpha-fetoprotein (AFP) were collected from each group, and ELISA was used to measure the serum level of ATG7. The receiver operating curve (ROC) was plotted for ATG7 and AFP used alone or in combination, and the area under the ROC curve (AUC) was compared. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups, and the Mann-Whitney U test was used for comparison between two groups; the chi-square test was used for comparison of categorical data between groups; a Spearman correlation analysis was used to investigate correlation. Results The serum level of ATG7 was 22.88(19.79-23.04) ng/mL in the HBV-HCC group, 17.06(14.45-19.40) ng/mL in the non-HBV-HCC group, 19.21(16.65-20.82) ng/mL in the CHB group, and 13.82(8.70-17.82) ng/mL in the HC group, with a significant difference between groups ( χ 2 =65.144, P < 0.001). ATG7 had an AUC of 0.818 (95% confidence interval [ CI ]: 0.743-0.879) and AFP had an AUC of 0.777 (95% CI : 0.698-0.843) in the diagnosis of HBV-HCC, suggesting that ATG7 had a slightly higher AUC than AFP ( Z =0.852, P =0.394). ATG7 combined with AFP had an AUC of 0.859 (95% CI : 0.790-0.913) in the diagnosis of HBV-HCC, which was significantly higher than the AUC of ATG7 alone ( Z =2.192, P =0.028) and AFP alone ( Z =2.076, P =0.038). Conclusion ATG7 is a good marker for the diagnosis of HBV-HCC, and combined measurement of ATG7 and AFP can significantly improve the diagnostic rate for HBV-HCC.

Objective To evalute the clinical application of the new technique of instantaneous wave intensity(WI) through the correlative analysis of instantaneous acceleration wave intensity(W1) and Tei index. Methods The correlation of the intensity of W1 (the apogee of W1) and Tei index of 66 healthy adults were analyzed by Prosound a10 color Doppler ultrasonograph. Results There were positive correlations in pressure and caliber between the common carotid arteries of both sides in 66 cases of healthy adults (P <0.01). There were negative correlations respectively between the pressure of W1 of left and right common carotid arteries and Tei index (P <0.05),and there were the same in caliber (P <0.01), mean pressure (P <0.05) and mean caliber (P <0.01). Conclusions W1 is negatively correlated with Tei index and can be an index in judging the systolic function.

Objective To identify the crude drugs of Anoectochilus burmannicus, and clarify its original plant pharmacognostical and microscopic characteristics. Methods The pharmacognostical identification method was used to observe the original plant, tissue structure and microscopic characteristics of A. burmannicus. Results Leaves were ovate or ovate elliptic with golden-red veins. Non-inverted yellow flowers had Y-shaped and yellow labellum, which were anteriorly enlarged and 2-lobed. The lobes were narrowly oblong or narrowly oblanceolate. The middle part of labellum was narrow to form a 10 mm long structure with margin narrowly winged. In the microscopic structure, the cortex is obvious in the cross sections of root and stem, together with needle crystals of calcium oxalate and mucous cells. The upper epidermal cells on the cross section of the leaves were papilloid in shape, whereas diverse stomas existed among the lower epidermal cells, with anomocytic stomas as the major type. Needle crystals of calcium oxalate and conduits can be found in the powder. Conclusion These data provide a reference for the identification and resource development and utilization of A. burmannicus.

Objective To perform the pharmacognostic identification of Anoectochilus lylei and establish the foundation for its accurate identification and further development. Methods The macroscopic identification and microscopic identification methods were used to identify A. lylei. Results A. lylei has ovate leaf shape, possessing red reticulated veins. Inverted flowers have Y-shaped and white lip. The anterior part of lip is two-lobed, and the lobes are linear-oblong. There are 1 to 3 shorts serrations on each side of the middle part of lip. Microscopic characteristics mainly show as follows: the cortex is broad in the transverse section of roots and stems; 1-5 and 1-7 vascular bundles in the xylem of transverse section of roots and stem, respectively. Collateral vascular bundle in the main veins of transverse section of leaves. There are multitudinous types of stomas in the leaf abaxial epidermis, most of which are anomocytic. Conclusion These characteristics could provide reference for the correct identification of A. lylei.

Objective: To investigate the effects of costunolide (Cos) on the proliferation, apoptosis, migration andinvasion of cholangiocarcinoma RBE cells, and explore its potential mechanism. Methods: The CCK-8, flow cytometry,Annexin V-FITC/PI double staining, Transwell assays were used to examine the influence of Cos on proliferation, cell cycle, apoptosis, migration and invasion of RBE cells after treated with gradient concentrations of Cos. The expressions of M M P 2 and M M P 9 were detected by qRT-PCR, and the expression of PI3K/AKT-associated signal proteins was detected by Western blotting. Results: Cosdose-dependently inhibited proliferation activity of RBE cells( P <0.05 or P <0.01), arrested cell cycle at S and G2/M phases and induced RBE cell apoptosis( P <0.01). Transwell and qRT-PCR results demonstrated that Cos impeded RBE cell migration, invasion, and reduced the transcription of M M P 2 and M M P 9 . Cos inhibited the expressionofp-AKT, Bcl-2, MMP2 and MMP9, the level of Bax. Conclusion: Cos restrained the proliferation, migration and invasion of RBE cells by suppressing PI3K/AKTpathway.

OBJECTIVE: To evaluate the effects of a 48-week course of adefovir dipivoxil (ADV) plus Chinese medicine (CM) therapy, namely Tiaogan Jianpi Hexue () and Tiaogan Jiedu Huashi () fomulae, in hepatitis B e antigen (HBeAg)-positive Chinese patients. METHODS: A total of 605 HBeAg-positive Chinese CHB patients were screened and 590 eligible participants were randomly assigned to 2 groups in 1:1 ratio including experimental group (EG, received ADV plus CM) and control group (CG, received ADV plus CM-placebo) for 48 weeks. The major study outcomes were the rates of HBeAg and HBV-DNA loss on week 12, 24, 36, 48, respectively. Secondary endpoints including liver functions (enzymes and bilirubin readings) were evaluated every 4 weeks at the beginning of week 24, 36, and 48. Routine blood, urine, and stool analyses in addition to electrocardiogram and abdominal B scan were monitored as safety evaluations. Adverse events (AEs) were documented. RESULTS: The combination therapy demonstrated superior HBeAg loss at 48 weeks, without additional AEs. The full analysis population was 560 and 280 in each group. In the EG, population achieved HBeAg loss on week 12, 24, 36, and 48 were 25 (8.90%), 34 (12.14%), 52 (18.57%), and 83 (29.64%), respectively; the equivalent numbers in the CG were 20 (7.14%), 41 (14.64%), 54 (19.29%), and 50 (17.86%), respectively. There was a statistically significant difference between these group values on week 48 (P<0.01). No additional AEs were found in EG. Subgroup analysis suggested different outcomes among treatment patterns. CONCLUSION Combination of CM and ADV therapy demonstrated superior HBeAg clearance compared with ADV monotherapy. The finding indicates that this combination therapy may provide an improved therapeutic effect and safety profile (ChiCTR-TRC-11001263).
Adenine ; analogs & derivatives ; therapeutic use ; Adult ; Antiviral Agents ; therapeutic use ; Double-Blind Method ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hepatitis B e Antigens ; immunology ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Male ; Medicine, Chinese Traditional ; Organophosphonates ; therapeutic use ; Young Adult