[Qualitative and quantitative analyses of Tripterygium hypoglaucum in Yinning Tablets, a compound traditional Chinese herbal preparation].

OBJECTIVE: To conduct qualitative and quantitative analyses of Tripterygium hypoglaucum in Yinning Tablets, a compound preparation of traditional Chinese herbal medicine. METHODS: Thin-layer chromatography (TLC) was used for qualitative analysis of Tripterygium hypoglaucum in Yining Tablets and the analytical protocols were optimized. High-performance liquid chromatography (HPLC) was used to quantitatively analyze the content of triptolide (the main active ingredient of Tripterygium hypoglaucum) in Yinning Tablets. RESULTS: The results of TLC analysis showed that the test sample of Yinning Tablets and the positive control samples both produced clear, well separated spots without obvious interference in the blank samples. Assessment of the influences of the thin-layer plates from different manufacturers, temperature and humidity on the test results demonstrated good durability of the test. HPLC analysis of triptolide showed a good linear relationship within the concentration range of 1-100 µg/mL (regression equation: A=22.219C-19.165, r=0.9999); the contents of triptolide in 3 batches of Yinning tablets were 0.34, 0.34, and 0.28 µg per tablet, all within the range of 0.28-0.34 µg per tablet. It was finally determined that each Yinning tablet should not contain more than 0.6 µg of triptolide. CONCLUSION: TLC and HPLC are simple, accurate, durable and specific for qualitative and quantitative analyses of Tripterygium hypoglaucum in Yinning Tablets.

LINC00205, a YY1-modulated lncRNA, serves as a sponge for miR-26a-5p facilitating the proliferation of hepatocellular carcinoma cells by elevating CDK6.

OBJECTIVE: LINC00205, a bidirectional lncRNA, located at human chromosome 21q22.3, was recently characterized as an oncogenic molecule contributing to cell proliferation in several cancers, including hepatocellular carcinoma (HCC). In the present study, we aim to probe the new molecular mechanism for LINC00205 controlling the proliferation of HCC cells. PATIENTS AND METHODS: The expression status of LINC00205, miR-26a-5p, as well as CDK6 in HCC tissues/cell lines was determined by quantitative real-time PCR (qPCR). The cell proliferative activity was measured by using the Cell Counting Kit (CCK)-8 assay. Flow cytometry was performed to analyze cell cycle progression and apoptosis induction. The interaction among LINC00205, miR-26a-5p and CDK6, as well as transcription efficiency of LINC00205 promoter were examined by Dual-Luciferase reporter assay. Western blot was conducted to evaluate the protein levels of CDK6 in SNU-449 cells. The direct interplay between YY1 and LINC00205 promoter was detected by ChIP-qPCR. RESULTS: LINC00205 was strongly expressed in HCC tissues and cell lines. Elevated LINC00205 expression was positively associated with worse prognosis as well as pathological grade in HCC. Suppression of LINC00205 could impede the proliferation of HCC cells by triggering the G0/G1-phase cell cycle arrest and apoptosis in vitro. Mechanistically, we illustrated that LINC00205 could accelerate the proliferation of HCC cells by boosting CDK6 expression via sponging miR-26a-5p. Moreover, we unveiled that LINC00205 could be activated by transcription factor Yin Yang-1 (YY1) as its direct downstream target. CONCLUSIONS: LINC00205, a novel YY1-modulated lncRNA, can facilitate the proliferation of HCC cells through YY1/miR-26a-5p/CDK6 pathway, and may serve as a promising diagnostic biomarker and therapeutic target for HCC.

Safety evaluation of zinc methionine in laying hens: Effects on laying performance, clinical blood parameters, organ development, and histopathology.

The study was conducted to investigate whether high-dose zinc methionine (Zn-Met) affected the safety of laying hens, including laying performance, hematological parameters, serum chemical parameters, organ index, and histopathology. A total of 540 20-week-old Hy-Line White laying hens was randomly allocated to 6 groups with 6 replicates of 15 birds each. Birds were fed diets supplemented with 0 (control), 70, 140, 350, 700, or 1,400 mg Zn/kg diet as Zn-Met. The experiment lasted for 8 wk after a 2-week acclimation period. Results showed that dietary supplementation with 70 or 140 mg Zn/kg diet as Zn-Met significantly increased average daily egg mass (ADEM), laying rate (LR), and feed conversion ratio (FCR) (P < 0.05) and lowered broken and soft-shelled egg ratio (BSER) (P < 0.05) in comparison with the control group; no significant differences were detected among hens fed with 0, 350, or 700 mg Zn/kg as Zn-Met (P > 0.05); hens administered 1,400 mg Zn/kg showed a significant increase in BSER and remarkable decreases in ADEM, LR, and FCR (P < 0.001). There were no significant differences among hens receiving 0, 70, 140, 350, or 700 mg Zn/kg as Zn-Met in serum chemical parameters (P > 0.05); supplementation with 1,400 mg Zn/kg as Zn-Met remarkably elevated the concentrations of serum total bilirubin (TBILI), glucose (GLU), uric acid (UA), and creatinine (CRE) (P < 0.001), and enhanced activities of serum glutamic oxalacetic transaminase (GOP) and alkaline phosphatase (AKP) (P < 0.001) compared with the control group. No significant histopathological changes were found in hens administered 0, 70, 140, 350, or 700 mg Zn/kg as Zn-Met, while significant histological lesions were observed in the heart, liver, lung, and kidney tissues of hens receiving 1,400 mg Zn/kg as Zn-Met. No significant differences were detected in hematological parameters or organ index (P > 0.05). In conclusion, a nominal Zn concentration of 700 mg/kg as Zn-Met is considered to be no-observed-adverse-effect level following daily administration to hens for 56 days.

The antimicrobial peptide sublancin ameliorates necrotic enteritis induced by Clostridium perfringens in broilers.

Sublancin is an antimicrobial peptide produced by 168 containing 37 amino acids. The objective of this study was to investigate its inhibitory efficacy against both in vitro and in vivo. In the in vitro study, we determined that sublancin had a minimum inhibitory concentration of 8 µM against , which was much higher than the antibiotic lincomycin (0.281 µM). Scanning electron microscopy showed that sublancin damaged the morphology of . The in vivo study was conducted on broilers for a 28-d period using a completely randomized design. A total of 252 chickens at 1 d of age were randomly assigned to 1 of 6 treatments including an uninfected control; an infected control; 3 infected groups supplemented with sublancin at 2.88, 5.76, or 11.52 mg activity/L of water; and an infected group supplemented with lincomycin at 75 mg activity/L of water (positive control). Necrotic enteritis was induced in the broilers by oral inoculation of on d 15 through 21. Thereafter, the sublancin or lincomycin were administered fresh daily for a period of 7 days. The challenge resulted in a significant decrease in ADG ( < 0.05) and a remarkable deterioration in G:F ( < 0.05) during d 15 to 21 of the experiment. There was a sharp increase of numbers in the cecum ( < 0.05). The addition of sublancin or lincomycin reduced caecal counts ( < 0.05). The counts had a tendency to decrease in the lincomycin treatment ( = 0.051) but were the highest in the sublancin treatment (5.76 mg activity/L of water). A higher villus height to crypt depth ratio in the duodenum and jejunum as well as a higher villus height in the duodenum were observed in broilers treated with sublancin or lincomycin ( < 0.05) compared with infected control broilers. It was observed that sublancin and lincomycin decreased IL-1ß, IL-6, and tumor necrosis factor-α levels ( < 0.05) in the ileum compared with the infected control. In conclusion, although sublancin's minimum inhibitory concentration is much higher than lincomycin in vitro, less sublancin is needed to control necrotic enteritis induced by in vivo than lincomycin. These novel findings indicate that sublancin could be used as a potential antimicrobial agent to control necrotic enteritis.

Enhancement of fermentation process in Pu-erh tea by tea-leaf extract.

Pu-erh tea is known as a fermented tea and longer storage enhances its flavor and taste. Recently, Aspergillus, Blastobotrys, and Streptomyces are found to play important roles in nutritional enhancement of Pu-erh tea by fermentation. Since water and temperature affect the microbial growth, we therefore explored the factors that might enhance the Pu-erh tea fermentation. The results showed that the addition of fresh tea-leaf extract (TLE) enhanced the withered tea fermentation (at 37 degrees C, 80 to 85% RH) as compared with the water only. Contents of statin, GABA, gallic acid, DPPH scavenging and polyphenol oxidase (PPO) activities were increased, whereas polyphenols and caffeine were decreased over 6 mo. TLE dose-dependently enhanced some of the qualities (that is, statin, PPO) of Pu-erh tea significantly as compared with the water only. The effect was related to the increase population of A. niger and A. carbonarius at 6 mo (from 7.6 +/- 1.2 x 10(1) and 3.2 +/- 1.3 x 10(1) to 3.1 +/- 1.2 x 10(6) and 2.4 +/- 1.1 x 10(5) colony forming units [CFU]/g, respectively). After drying process (90 degrees C, 30 min), the total microbial count from these samples returned to background level (3 +/- 0.5 x 10(2) CFU/g). None of ochratoxin and fumonisin, toxins from Aspergillus, was detected in the final products. The flavor and taste were also enhanced by treatment with TLE. The inoculation with S. cinereus Y11 with 2% TLE further enhanced these functional contents (about 2-fold increase of statin level) in the experimental Pu-erh tea. Therefore, this result may add a new process for Pu-erh tea manufacture.

Inhibitory effect of dry needling on the spontaneous electrical activity recorded from myofascial trigger spots of rabbit skeletal muscle.

OBJECTIVE: Dry needling of myofascial trigger points can relieve myofascial pain if local twitch responses are elicited during needling. Spontaneous electrical activity (SEA) recorded from an active locus in a myofascial trigger point region has been used to assess the myofascial trigger point sensitivity. This study was to investigate the effect of dry needling on SEA. DESIGN: Nine adult New Zealand rabbits were studied. Dry needling with rapid insertion into multiple sites within the myofascial trigger spot region was performed to the biceps femoris muscle to elicit sufficient local twitch responses. Very slow needle insertion with minimal local twitch response elicitation was conducted to the other biceps femoris muscle for the control study. SEA was recorded from 15 different active loci of the myofascial trigger spot before and immediately after treatment for both sides. The raw data of 1-sec SEA were rectified and integrated to calculate the average integrated value of SEA. RESULTS: Seven of nine rabbits demonstrated significantly lower normalized average integrated value of SEA in the treatment side compared with the control side (P < 0.05). The results of two-way analysis of variance show that the mean of the normalized average integrated value of SEA in the treatment group (0.565 +/- 0.113) is significantly (P < 0.05) lower than that of the control (0.983 +/- 0.121). CONCLUSIONS: Dry needling of the myofascial trigger spot is effective in diminishing SEA if local twitch responses are elicited. The local twitch response elicitation, other than trauma effects of needling, seems to be the primary inhibitory factor on SEA during dry needling.

Electrophysiologic characteristics of ventricular extrastimulation-induced dissipation of functional bundle branch block associated with supraventricular tachycardia.

INTRODUCTION: Linking-related anterograde functional bundle branch block during supraventricular tachycardia (SVT) is due to repetitive concealed retrograde conduction of impulses from the contralateral bundle branch and can be eliminated by a critically timed premature ventricular beat (PVB). We assessed the electrophysiologic characteristics of PVB-induced dissipation of functional bundle branch block during SVT. METHODS AND RESULTS: During SVT with functional bundle branch block, PVB was delivered from the right ventricular apex, scanning the tachycardia cycle length (CL) with 10-msec decrements in the coupling interval in 14 patients (3 AV nodal reentrant tachycardia and 11 orthodromic AV reciprocating tachycardia). Dissipation was achieved in group 1: functional right bundle branch block (RBBB) in 4, functional left bundle branch block (LBBB) in 4, and both functional RBBB and LBBB in 1 with a dissipation zone occupying 4% to 13% (mean 8.5%) of the tachycardia CL. The outer limits were 22+/-16 msec and 68+/-14 msec < tachycardia CL; the inner limits were 56+/-18 msec and 90+/-24 msec < tachycardia CL for RBBB and LBBB, respectively (both P < 0.05). Dissipation could not be achieved in group 2 (4 RBBB and 1 LBBB) due to CL-dependent bundle branch block and/or local ventricular refractoriness. CONCLUSION: During SVT, functional bundle branch block due to "linking" often can be dissipated by timely PVB delivered from the right ventricular apex within a narrow zone of the tachycardia CL. Our findings suggest that the dissipation zone is affected by the pattern of functional bundle branch block relative to the site of PVB delivery.

[Experimental study on combining selective rhizotomy of different anterior and posterior sacral roots for restoration of bladder function after spinal cord injury].

OBJECTIVE: To investigate an alternative procedure for complete denervation of bladder in the supra-cone cord injury to restore the bladder function. METHODS: Sixteen dogs were included in this study after their spinal cords were transected above the cone. They were divided into 6 groups and performed the rhizotomy of L7 to S3 root in different combination respectively. The bladder and urethra pressure change by electrostimulation during operation and cystometrogram change after operation were tested. RESULTS: 1. Electrostimulation study: for bladder innervation, S2 was the most important and S1 was secondary. While for urethra innervation, S1 was more important than S2. When the anterior and posterior roots of S1 and S2 were intact with rhizotomy of posterior roots of L7 and S3, stimulated the common or posterior root of S1 and S2, the change of pressure in bladder and urethra was the same. When the anterior roots of S1 and S2 were resected with rhizotomy of posterior roots of L7 and S3, the pressure in bladder and urethra was significant decreased compared to stimulating the corresponding posterior roots. 2. Cystometrogram (CMG) study: in the complete deafferented group, resecting the posterior roots of L7 to S3, the bladder became flaccid. While resecting the posterior root of S2 and anterior root of S1 or, resecting the posterior root of S1 and anterior root of S2, combining with rhizotomy of posterior roots of L7 and S3, the CMG curve was similar to the complete deafferented group. In the S1 and S2 intact group, the bladder became spastic. CONCLUSION: Combining rhizotomy of anterior and posterior sacral root in different level has the same effects on bladder as complete deafferentation.

[Imitative restenosis after percutaneous transluminal coronary angioplasty prevented by buyang huanwu decoction in rabbits].

OBJECTIVE: To examine the preventive effect of Buyang Huanwu Decoction (BYHWD) on imitative restenosis after percutaneous transluminal coronary angioplasty (PTCA) in diet-induced atherosclerotic rabbits. METHODS: Diet-induced atherosclerotic rabbits were randomly allocated into group 1 (n = 24, fed with BYHWD one week before and 4 weeks after the operation, 2 ml/kg per day, containing herbal drug 8.4 g/ml) and group 2 (n = 20, as control). Balloon angioplasty was performed in the abdominal arota to mimic coronary angioplasty. RESULTS: Sixty days after operation: (1) Incidence of hyperplasia of the intima was 7/24 (29.2%) in group 1 vs 17/20 (85.0%) in group 2, P < 0.01; (2) Comparison of area of hyperplastic intima was (0.54 +/- 0.21) mm2 vs (1.67 +/- 0.75) mm2, P < 0.01; (3) Area of hyperplastic intima/area of lumen (limited by the internal elastic membrane was (18.0 +/- 7.2)% vs (56.0 +/- 17.1)%, P < 0.01; (4) Intact lumen/dilated lumen was (0.74 +/- 0.03) vs (0.35 +/- 0.07), P < 0.01; (5) Effect of serum taken from group 1 on cultured smooth muscle cells showed by 3H-TdR method was (4110.14 +/- 1977.01) cpm, which was significantly less than that of serum taken from group 2 [(7960.14 +/- 2802.59) cpm, P < 0.05]. CONCLUSION: BYHWD has significant preventive effect on the imitative post PTCA restenosis in diet-induced atherosclerotic rabbits.

Is serum transferrin receptor a sensitive marker of iron repletion in patients with iron-deficiency anemia and hemodialysis patients?

BACKGROUND: Serum transferrin receptor (sTfR) concentration has been recognized as a reliable laboratory indicator of iron deficiency in recent years. But its response to iron supplementation has not been investigated. METHODS: We evaluated the sTfR concentrations in 15 patients diagnosed with iron-deficiency anemia, in 30 patients receiving maintenance hemodialysis (HD) with iron repletion and in 31 healthy controls. The serial changes of sTfR concentration and their correlation with serum ferritin in patients with iron deficiency under iron repletion were also examined in three patients. RESULTS: In patients with iron-deficiency anemia, the sTfR concentration was 5.6 +/- 2.4 mg/ml, significantly higher than that in the control group (1.8 +/- 0.4 mg/ml) and patients receiving maintenance HD with iron repletion (1.7 +/- 0.5 mg/ml). The three patients with iron-deficiency anemia who received eight to 16 weeks of iron supplementation showed steady and significant decreases in sTfR concentration and significant increases in serum ferritin and transferrin saturation. However, the decreases in sTfR concentration did not occur immediately, as did the increases in serum ferritin and transferrin saturation, following iron repletion. There was a four-week delayed response in the decrease of sTfR concentrations as measured against serum ferritin and transferrin saturation. CONCLUSIONS: sTfR concentration may not be as effective as an early index of iron repletion compared with serum ferritin and transferrin saturation.

[Studies on the ganoderic acid, a new constituents from the fruiting body of Ganoderma lucidum (Fr.) Karst].

Three compounds have been isolated from the dichloromethane soluble fraction of the fruiting body of Ganoderma lucidum (Fr.) Karst. On basis of spectral analyses (UV, IR, MS, 1HNMR, 13CNMR and 2D-NMR), they were identified as 3, 7-dioxo-lanosta-8, 24(E)-dien-26-oic acid (I), 7 beta-15 alpha-dihydroxy-3, 11, 23-trioxo-5 alpha-lanost-8-en-26-oic acid (II) and 3 beta, 7 beta, 15 alpha-trihydroxy-11, 23-dioxo-5 alpha-lanosta-8-en-26-oic acid (III). Compound I is a new compound named ganoderic acid DM.

[Isoflavones from Glycyrrhiza eurycarpa].

A new isoflavone, named eurycarpin A and a new natural product isoflavone named eurycarpin B have been isolated from the roots of Glycyrrhiza eurycarpa P. C. Li. Their structures were determined to be 7,2',4'-trihydroxy-3'-(3,3-dimethylallyl) isoflavone(I) and 7,2'-dihydroxy-6",6"-dimethylpyrano-(2",3":4',3') isoflavone(II) on the basis of spectroscopic analysis (UV, EI-MS, 1HNMR, 13CNMR, NOE difference and HMBC). In addition, three known isoflavones, licoisoflavone A, calycosin and formononetin, were obtained for the first time from this plant.

[Chemical constituents of the roots of Glycyrrhiza eurycarpa P.C.Li].

Four compounds were isolated from the roots of Glycyrrhiza eurycarpa. They were identified as licochalcone A, glabrol, betulic acid and beta-sitosterol by chemical and physical properties and spectral analysis.

[Study on platelet-derived growth factor mRNA and copper-zinc superoxide dismutase mRNA expression changes of aortic artery endothelium with restenosis after aortic artery dilatation treated with buyang huanwu decoction].

OBJECTIVE: To study on dynamic mechanism of restenosis after PTCA treated with Buyang Huanwu Decoction (BYHWD). METHODS: Twenty-eight male New Zealand rabbits with mimic restenosis after aortic artery dilatation were detected by prime in situ labeling in aortic endothelium. The items included gene expression of copper zinc superoxide dismutase (SOD-1) and platelet-derived growth factor receptor (PDGFR) mRNA to analyse dynamic mechanism of restenosis. RESULTS: Seven days after aortic artery dilatation, PDGFR mRNA of aortic endothelium decreased clearly. The comparison between experimental and the control group showed significant difference (P < 0.05). By the same condition, the gene expression of SOD-1 was increasing, which showed a significant difference with the control group (P < 0.05). CONCLUSIONS: BYHWD increased SOD-1 gene expression of artery endothelium, to improve oxygen feed for ischemia reperfusion in the same way, BYHWD suppressed PDGFR mRNA expression to control cellular pathological proliferation.

[Chemical constituents of Swertia pubescens Franch].

Four compounds were isolated from Swertia pubescens. They were identified as isoorientin, gentiopicroside, glucose and oleanolic acid by chemical and physical properties and spectral analysis.

[In vitro experiment on scavenging oxy-radical and suppressing myocardial lipid peroxidation with sini decoction].

Results indicate that Sini Decoction has a significant effect of anti-lipid peroxidation of its components, Radix Aconiti Preparata and Radix Glycyrrhizae can significantly suppress lipid peroxidation of myocardial homogenate, while Rhizoma Zingiberis cannot. The decoction can effectively scavenge oxygen free radicals, each component having similar effect to some extent.