Objective: To establish an HPLC fingerprint method of Verbenae officinalis L. and determine the contents of its main components. Methods: Many batches of Verbenae officinalis fingerprint and the contents of main components were determined by HPLC, and the similarity and cluster analysis of the fingerprints of each batch were compared and the contents of the main components were compared. Results: The results showed that the difference of the main content of medicinal materials and fingerprints were between each batch of Verbenae officinalis. Conclusion: The fingerprint and its main components determination method have been established to provide the scientific basis for the quality evaluation of Verbenae officinalis.

Objective: To establish the HPLC fingerprint of herbs of Patriniae Herba. Methods: The fingerprints of Patriniae Herba were built using Orca C18 column and acetonitrile-0.05% phosphoric acid as mobile phase. The flow rate was 1.0 mL/min. The detecting wavelength was set at 230 nm. The temperature of column was at 35℃. Results: Under the selected spectrum conditions, HPLC fingerprint of herbs of Patriniae Herba was established, and eight public peaks were shown in the HPLC fingerprint. The methodological study met the required standards. Cluster analysis showed that class I medical BJ1, BJ2, BJ3, BJ4, BJ5, BJ6, BJ7, BJ8, BJ9, both, BJ13, BJ14 each batch Patriniae Herba and control the similarity between fingerprints was 0.987-0.907, Indicating that there are good consistency between batches of medicinal materials; class II medical BJ11, BJ12, and BJ15 group of Patriniae Herba and control fingerprint differences larger. Conclusion: The method is accurate and reliable, simple and efficient, and can be used as the evaluation of Patriniae Herba from different origins and with different varieties.

Aim To investigate the effects of diallyl di-sulfide( DADS) on G2/M arrest in Chk1/MGC803 and Chk2/MGC803 cells so as to establish stable human gastric cancer MGC803 cells with overexpression of Chk1/2 gene. Methods The colony formation, flow cytometry, RT-PCR and Western blot were used to de-tect the proliferation, cell cycle, and expression of Chk1/2 mRNA and protein, p-Chk1/2, CDC25C and cyclinB1, respectively. Results The colony formation showed that the colony forming efficiency in Chk1/MGC803 and Chk2/MGC803 cells treated by 30 mg· L-1 DADS was lower than in control group and vector group ( P <0. 05 ) . Flow cytometry demonstrated that 41. 3%, 57. 4%, 68. 9% and 42. 9% of G2/M cells in Chk1/MGC803 were increased than in MGC803 and Chk2/MGC803 , respectively after treated by DADS in 12,24, 36 and 48 h(P <0. 05). At the same time, RT-PCR disclosed that expression of Chk1 and Chk2 mRNA had no marked change. Western blot showed that total proteins of Chk1 and Chk2 and p-Chk2 had invisible change, but expression of p-Chk1 was up-reg-ulated, and CDC25C and cyclinB1 were down-regula-ted time-dependently in Chk1/MGC803 cells ( P <0. 05 ) . Conclusion DADS arrests MGC803 cells at G2/M by increasing p-Chk1 expression to cause down-regulation of CDC25C and cyclinB1 simultaneously.

Objective To optimize purification technology of total flavonoids in Callicarpa nudiflora Hook. et Arm. by macroporous adsorption resin. Methods Macroporous resin models including AB-8, D-101, HP-20, HP2MG, were optimized by static adsorption and desorption experiments regarding to adsorption rate and desorption rate of total flavonoids. Purification technology parameters of total flavonoids were optimized by single factor test. Results HP-20 macroporous resin presented the best purification efficiency,the optimum purification conditions were that taking 4. 46 mg·mL-1 of total flavonoidsat pH 3. 0, loading at 3 BV·h-1, washed with 3BV of water at 3 BV·h-1,then eluted with 4 BV 75% ethanol at 2 BV·h-1, finally obtaining the total flavonoids from the dry extract of Callicarpa nudiflora Hook. et Arn. with the purity of 47. 4%. Conclusion HP-20 macroporous resin is suitable for preliminary purification of total flavonoids in Callicarpa nudiflora Hook. et Arn.

To observe BAG-1, EGFR, and PARP-1 expressions in invasive breast cancer and its correlation with clini-cal pathological indicators, as well as to evaluate their clinical significance. Methods:The BAG-l, EGFR, and PARP-1 expressions in a tissue microarray of invasive breast cancer and peritumoral tissues were detected through immunohistochemical staining. The clinical and pathological significance of BAG-1, EGFR, and PARP-1 were evaluated. Results:The BAG-1, EGFR, and PARP-1 expression lev-els are higher in invasive breast cancer tissues than in peritumoral tissues (P<0.05). BAG-1 expression in invasive cancer tissues is not related to age, tumor site, lymph node metastases, and clinical TNM staging of patients, but is related to size, grade, ER, PR, and HER-2 expressions and molecular subtype (P<0.05). EGFR expression is related to size, clinical TNM staging, and molecular subtype (P<0.05). PARP-1 expression is related to grade, lymph node metastases, ER, and molecular subtype (P<0.05). BAG-1 expression is not significantly correlated with EGFR and PARP-1 in all cases, but BAG-1 and PARP-1 expressions are positively correlated in tri-ple-negative breast cancer tissues (P<0.05). Results of the univariate analysis revealed that the BAG-1 and PARP-1 expressions and the molecular subtypes are associated with the prognoses of breast cancer patients. Multivariate analysis revealed that BAG-1 and PARP-1 expressions are factors that are independent of the prognosis. Conclusion: BAG-1, EGFR, and PARP-1 overexpressions in human breast tissues suggest that BAG-1, EGFR, and PARP-1 are related to breast cancer development. BAG-1, EGFR, and PARP-1 are poten-tial biomarkers of breast cancer diagnosis and prognosis.

AIM:To research the effects of lithium chloride on transforming growth factor beta (TGF-β) and connective tissue growth factor (CTGF) in cultured human Tenon capsule fibroblasts (HTFs) and explore its mechanism.METHODS:HTFs were cultured and identified by vimentin staining with immunofluorescence and the morphological characteristics.The experimental group was processed 48h with LiCl in concentration of 80mmol/L, the control group without LiCl.The mRNA expression of TGF-β and CTGF in two groups were analyzed with real-time fluorescent quantitative polymerase chain reaction (real time-qPCR) and the protein expression was detected with Western blot.RESULTS:The cultured HTFs expressed TGF-β and CTGF.The mRNA expression of TGF-β and CTGF significantly decreased compared with the control group(t=20.042, 14.995, P<0.05).the protein expression of TGF-β and CTGF also decreased significantly compared with the control group(t=46.058、12.452, P<0.05)CONCLUSION:The cultured HTFs can express TGF-β and CTGF in mRNA and proteins' level.LiCl can reduce the expression of TGF-β and CTGF both in gene and proteins' level.LiCl has the potential to modulate wound healing for glaucoma filtration surgery.

This article discussed ZHU Lian's contributions to acupuncture-moxibustion scientific research from three aspects: building the scientific thought of "new acupuncture-moxibustion", constructing the first domestic acupuncture-moxibustion institution and opening the door to modern acupuncture-moxibustion scientific research. ZHU Lian's visionary thought of "new acupuncture-moxibustion" has influenced the following researchers till now. She established the acupuncture-Moxibustion therapeutic institute affiliated to the Ministry of Health, set up the acupuncture-Moxibustion research platforms and teams and made research cooperation. She firstly carried out acupuncture-Moxibustion clinical and basic scientific research, which started the acupuncture-Moxi- bustion scientific research in China. ZHU Lian is the Pioneer of Chinese acupuncture-Moxibustion scientific research.
Acupuncture ; education ; Acupuncture Therapy ; history ; China ; History, 20th Century ; Humans ; Moxibustion ; history

AIMTo explore the modulation of Salvia miltiorrhiza on hyperpolarization-activated current (Ih) channels in dorsal root ganglion (DRG) neurons of rats and identify the mechanism of Salvia miltiorrhiza in alleviating pain and inhibiting calcium overload. Methods The effect of Salvia miltiorrhiza injection on Ih channels in DRG neurons of rats were examined by using whole-cell patch clamp technique. Results The experimental results showed that the amplitude of Ih evoked by -150 mV was (-1.06 +/- 0.18) nA. The Ih could be fitted well into the single kinetics and the time constant of activation, pi was clearly voltage-dependent with tau = (322.14 +/- 28.81) ms at -100 mV, decreasing to tau = (62.51 +/- 9.78) ms at -150 mV. The reversal potential of Ih was (-35.03 +/- 1.12) mV measured from tail currents. But no significant differences were found between the DRG neurons in the absence and presence of Salvia miltiorrhiza injection (10%, 25%, 50%) in the current amplitude, the time constant of activation and the reversal potential. The only difference between the DRG neurons in the absence and presence of Salvia miltiorrhiza injection was the half-activation potential of Ih. In control recordings the half-activation potential was (-106.07 +/- 3.59) mV. By comparison, the half-activation potentials changed to (-111.59 +/- 3.79) mV (n=31 neurons, P < 0.05), (-119.37 +/- 4.96) mV (n=31 neurons, P < 0.05) and (-121.23 +/- 3.86) mV (n=31 neurons, P < 0.05) in the presence of 10%, 25%, 50% Salvia miltiorrhiza injection, respectively.

CONCLUSIONOnly the half-activation potential of Ih in the arthritic and neuropathic rat models shifted in the depolarizing direction, which increased the electrophysiological activity of Ih and made it related to peripheral hyperalgesia. The selective inhibition of Salvia miltiorrhiza on the electrophysiological activity of Ih may be one of the mechanisms underlying its analgesic effects.

Action Potentials ; drug effects ; Analgesics, Non-Narcotic ; administration & dosage ; isolation & purification ; pharmacology ; Animals ; Cells, Cultured ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Ganglia, Spinal ; cytology ; Injections ; Ion Channels ; physiology ; Neurons ; cytology ; drug effects ; physiology ; Patch-Clamp Techniques ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Salvia miltiorrhiza ; chemistry

Paeoniflorin is the main active ingredient of Chinese herbaceous peony. This study is to investigate the protective effect of paeoniflorin (Pae) on acute brain damage induced by lipopolysaccharide (LPS) in mice. The mice were randomly assigned to the normal control, model control (LPS), as well as groups of paeoniflorin and lipopolysaccharide (Pae + LPS). Then the mice were administered intraperitioneally with normal saline or Pae (10, 30 mg · kg(-1)) once daily for 6 d. One hour after intrapertioneally treatment on the seventh day, each group were injected LPS (5 mg · kg(-1)) to establish the endotoxin lipopolysaccharide inflammation model except the normal group. The mice were sacrificed after 6 h and the brain homogenates were prepared and measured. The malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2), succinatedehydrogenase (SDH), Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase were dectected by the colorimetric method. The levels of HO-1 and Nrf2 protein in subcellular fractions of brain tissue were detected by Western blot. The results demonstrated that the administration with paeoniflorin reduced the levels of the MDA production; significantly increase the activities of antioxidant enzyme (SOD and GSH-PX). In addition, paeoniflorin could enhance the total antioxidant capacity, decrease the level of H2O2, and increase the activities of SDH, Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase. Furthermore, paeoniflorin can increase the expression of HO-1 and activate the nuclear transfer of Nrf2. Taking together, these findings suggest that paeoniflorin alleviate the acute inflammation in mice brain damage induced by LPS, which is related with its antioxidant effect and improvement of energy metabolism.
Animals ; Energy Metabolism ; drug effects ; Glucosides ; pharmacology ; Heme Oxygenase-1 ; genetics ; Lipopolysaccharides ; pharmacology ; Male ; Membrane Proteins ; genetics ; Mice ; Mice, Inbred BALB C ; Monoterpenes ; pharmacology ; Oxidative Stress ; drug effects ; Sodium-Potassium-Exchanging ATPase ; metabolism

Objective To investigate the clinical diagnostic value and pathogenesis of serum protein identification in Keshan disease (KD).Methods A total of 65 chronic KD patients were selected as the patient group in KD endemic areas,while 29 cases of dilated cardiomyopathy (the DCM group),62 healthy cases from KD endemic areas (control 1 group) and 28 healthy cases from non-endemic areas (control 2 group) were selected as controls.Liquid chip time of flight mass spectrometry (ClinProtTM MALDI-TOF-MS) was used to determine the expression of proteins/peptide peaks.ClinProTools 2.2 software was used to analyze the protein profiles to determine differentially expressed proteins/peptide peaks.The Genetic Algorithm (GA),QuickClassifer Algorithm (QC) and Supervised Neural Network Algorithm (SNN) methods were used to screen marker proteins.Matrix-assisted laser desorption/ionization time-of-flight Mass Spectrometry technique (MALDI-TOF/TOF) was also used as a secondary mass spectrometry to identify differentially expressed peptides.Results Between the KD and control 1 groups,34 differentially expressed proteins/peptides and 5 marker proteins were identified,while 52 differentially expressed proteins/peptides and 5 marker proteins were identified between the KD and control 2 groups,and there were 67 differentially expressed proteins/peptides and 5 marker proteins between the KD and DCM groups.During secondary mass spectrometry,two peptides for mass-to-charge ratio (m/z) 2 079 and 1 465 were obtained,peptide of matching β-globin showed low expression while peptide of matching fibrinogen showed high expression in the KD patients.Conclusions Serum marker proteins can be used as biomarkers for diagnosis and differentiation of KD.β-globin and fibrinogen play an important role in the development of KD myocardial injury.