New steroidal saponins from the aerial parts of Paris polyphylla var. yunnanensis and their effects on blood coagulation.

Five new steroidal saponins, paripolins D-H (1-5), and 6 known compounds (6-11) were isolated from the aerial parts of Paris polyphylla var. yunnanensis. The structures of 1-5 were determined using spectroscopic analyses in conjunction with acid hydrolysis. It is for the first time to report the 12-hydroxysteroidal saponins from the genus Paris. The effect of all isolated compounds on blood coagulation was determined in vitro using the plasma recalcification time method. Compounds 1 and 2 showed potent procoagulant activity, and 5-11 exhibited significant anticoagulant activity.

Effects of Sensitized Sorafenib with a Paeoniflorin and Geniposide Mixture on Liver Cancer via the NF-κB-HIF-2α-SerpinB3 Pathway.

Purpose: This study focused on determining the anticancer effect of paeoniflorin and geniposide mixture (PFGS) combined with sorafenib (Sor) in hepatocellular carcinoma (HCC) and, in particular, whether PFGS increases the antitumor effect of Sor by modulating the NF-κB/HIF-2α/SerpinB3 pathway. Methods: The H22 hepatoma tumor-bearing mouse model was treated with PFGS, Sor, and a combination of the two drugs for 12 days. The effects of PFGS combined with Sor on tumor growth and apoptosis and the expression of NF-κB, HIF-2α, and SerpinB3 in tumor tissue were assessed. In addition, Sor-resistant hepatoma cells were treated with PFGS, Sor, and the combination of the two drugs in vitro. The effects of PFGS combined with Sor on cell proliferation and invasion and the protein expression of NF-κB p65, HIF-2α, and SerpinB3 were investigated. Results: PFGS combined with Sor treatment synergistically inhibited tumor growth in HCC tumor-bearing mice. Immunostaining showed that PFGS combined with Sor treatment significantly decreased the expression of Ki-67 and obviously induced apoptosis in the tumor compared with a single treatment. Similarly, PFGS combined with Sor treatment significantly downregulated the expression of NF-κB, HIF-2α, and SerpinB3 in the tumor compared with a single treatment. Additionally, PFGS combined with Sor markedly inhibited cell proliferation and invasion and activation of the NF-κB/HIF-2α/SerpinB3 pathway in Sor-resistant hepatoma cells compared with a single treatment. Conclusion: Our study demonstrated that PFGS synergistically increased the antiliver cancer effects of Sor by lowering activation of the NF-κB/HIF-2α/SerpinB3 pathway. These findings provided a scientific foundation for clinical studies using PFGS and Sor to treat liver cancer.

Matrine-Family Alkaloids: Versatile Precursors for Bioactive Modifications.

Matrine-family alkaloids as tetracycloquinolizindine analogues from Traditional Chinese Medicine Sophora flavescens Ait, Sophora subprostrata and Sophora alopecuroides L possess various pharmacological activities and have aroused great interests over the past decades. Especially, a lot of matrine derivatives have been designed and synthesized and their biological activities investigated, and encouraging results have continuously been achieved in recent several years. These studies are helpful to develop more potent candidates or therapeutic agents and disclose their molecular targets and mechanisms. This paper reviews recent advances in the bioactive modifications of matrine-family alkaloids from derivatization of the C-13, C-14 or C-15 position, opening D ring, fusing D ring and structural simplification.

Nanoparticle based bio-bar code technology for trace analysis of aflatoxin B1 in Chinese herbs.

A novel and sensitive assay for aflatoxin B1 (AFB1) detection has been developed by using bio-bar code assay (BCA). The method that relies on polyclonal antibodies encoded with DNA modified gold nanoparticle (NP) and monoclonal antibodies modified magnetic microparticle (MMP), and subsequent detection of amplified target in the form of bio-bar code using a fluorescent quantitative polymerase chain reaction (FQ-PCR) detection method. First, NP probes encoded with DNA that was unique to AFB1, MMP probes with monoclonal antibodies that bind AFB1 specifically were prepared. Then, the MMP-AFB1-NP sandwich compounds were acquired, dehybridization of the oligonucleotides on the nanoparticle surface allows the determination of the presence of AFB1 by identifying the oligonucleotide sequence released from the NP through FQ-PCR detection. The bio-bar code techniques system for detecting AFB1 was established, and the sensitivity limit was about 10-8 ng/mL, comparable ELISA assays for detecting the same target, it showed that we can detect AFB1 at low attomolar levels with the bio-bar-code amplification approach. This is also the first demonstration of a bio-bar code type assay for the detection of AFB1 in Chinese herbs.

Glycemic index, glycemic load, and glycemic response to pomelo in patients with type 2 diabetes.

Food intake has a great influence on blood glucose in patients with diabetes. This study was to determine the glycemic index (GI) and glycemic load (GL) of a particular pomelo named Majia pomelo and its effects on postprandial glucose (PPG) in patients with type 2 diabetes (T2D). Twenty healthy subjects and 20 T2D patients (controlled on lifestyle measures and/or metformin) were tested on 2 separate days with 50 g of glucose and 50 g equivalent of carbohydrates from Majia pomelo for GI measurement. To test effects of Majia pomelo on PPG, 19 hospitalized T2D patients (controlled on insulin therapy) were selected for a 9-day study. The dose of insulin for each patient was adjusted on the first 3 days. A total of 100 mg Majia pomelo was consumed per meal in the last 3 tested days. Blood glucose was measured to evaluate the glycemic excursions. The GIs for Majia pomelo in healthy individuals and T2D patients were 78.34±1.88 and 72.15±1.95 respectively. The value of GL was as low as 4.23 in diabetic patients with serving size of 100 g pomelo, indicting Majia pomelo as a high GI but low GL fruit. Consumption of Majia pomelo in hospitalized T2D patients did not cause significant glucose fluctuation. It was concluded that high GI pomelo can serve as a low GL fruit if it is consumed with a limited daily amount and thus can be supplied to diabetic patients. These results may mean more varieties of food choices for T2D patients.

Qualitative analysis of a sulfur-fumigated Chinese herbal medicine by comprehensive two-dimensional gas chromatography and high-resolution time of flight mass spectrometry using colorized fuzzy difference data processing.

OBJECTIVE: To investigate the chemical transformation of volatile compounds in sulfur-fumigated Radix Angelicae Sinensis. METHODS: A comprehensive two-dimensional gas chromatography (GC×GC) and high-resolution time-of-flight mass spectrometry (HR-TOF/MS) with colorized fuzzy difference (CFD) method was used to investigate the effect of sulfur-fumigation on the volatile components from Radix Angelicae Sinensis. RESULTS: Twenty-five compounds that were found in sun-dried samples disappeared in sulfur-fumigated samples. Seventeen volatile components including two sulfur-containing compounds were newly generated for the first time in volatile oils of sulfur-fumigated Radix Angelicae Sinensis. CONCLUSION: The strategy can be successfully applied to rapidly and holistically discriminate sun-dried and sulfur-fumigated Radix Angelicae Sinensis. GC×GC-HR-TOF/MS based CFD is a powerful and feasible approach for the global quality evaluation of Radix Angelicae Sinensis as well as other herbal medicines.

[Effect of Picroside II on ERK1/2 Signal Pathway in Cerebral lschemic Injury Rats].

OBJECTIVE: To explore the neuroprotective effect and mechanism of picroside II on extracellular regulated protein kinases1/2 (ERK1/2) signal transduction pathway in cerebral ischemia injuryrats. METHODS The middle cerebral artery occlusion (MCAO) model was established by inserting a monofilament into middle cerebral artery. Totally 96 successfully modeled Wistar rats were divided into the modelgroup, the treatment (picroside II) group, the Lipopolysachcaride (LPS) group, and the U0126 group according to random digit table. Each group was further divided into 3 subgroups, i.e. 6, 12, and 24 h sub-groups. Picroside II (20 mg/kg) was peritoneally injected to rats in the treatment group 2 h after ischemia.LPS (20 mg/kg) and Picroside II (20 mg/kg) were peritoneally injected to rats in the LPS group 2 h after ischemia. U0126-EtOH (20 mg/kg)and Picroside II (20 mg/kg) were peritoneally injected to rats in the U0126group 2 h after ischemia. Equal volume of normal saline was peritoneally injected to rats in the control groupand the model group. The neurobehavioral function was evaluated by modified neurological severity score(mNSS) test. The structure of neurons was observed using hematoxylin-eosinstaining (HE) staining. Theapoptotic cells were detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The expression of phosphorylated extracellular signal-regulated protein kinase1,2 (pERK1,2) in cortex was detected using immunohistochemistry (IHC) and Western blot. RESULTS: After cerebral ischemia injury neurological impairment score increased, the damage of neuron in the cortical area was aggravated, apoptotic cells increased in the model group as time went by. The expression of pERK1/2 increased more significantly in the model group than in the control group (P <0.05). The damage of neuron in the cortical area was milder, while apoptotic cells decreased, the expression of pERK1f2 obviously decreased more in the treatment group and the U0126 group (P < 0.05). The early damage of neuron in the cortical area was more severe, apoptotic cells and the expression of pERK12 were comparatively higher in early stage of the LPS group, but the expression of pERK1/2 was somewhat decreased in late stage. CONCLUSIONS: Activating ERK12 pathway could mediate apoptosis and inflammatory reactions of neurons after cerebral ischemia injury. Picroside II could protect the nerve system possibly through reducing activation of ERKI2 pathway, inhibiting apoptosis of neurons and inflammation reaction.

[Bibliometric Analysis of Applying Proteomics Technology in Acupuncture and Moxibustion Re- search].

OBJECTIVE: To analyze document features and current research status of applying proteomics technology in acupuncture and moxibustion research, thereby further providing reference for further research and development in this area. METHODS: CNKI, Wangfang Data, CBM, PubMed, and Medline were included. Retrieved were literatures involving applying proteomics technology in acupunc- ture and moxibustion from January 2004 to December 2015. Document features and current research features were statistically analyzed using bibliometrics. RESULTS: Totally 91 articles were included, 60 from journals, 8 conference theses, and 23 dissertations. The 60 articles were published in 47 kinds of journals, and 56 of them were published in core journals. Theories of acupuncture and moxibustion (correlated with meridians, zang-fu organs) and mechanism research were hotspots, involving 19 kinds of disease models (dominated as gastric mucosal lesion, cerebral ischemia, and spinal cord injury). Two- dimensional gel based electrophoresis technology was used in 51 articles (69. 86%). CONCLUSIONS: Proteomics technology has been widely applied in the field of acupuncture and moxibustion research. It could promote researches on meridians and zang-fu organs correlated theories and mechanisms. The depth and breadth of the study is worth further widening.

The inhibitory effect of Shaoyao Ruangan formula on mice with transplanted H22 hepatocarcinoma and its mechanism research.

BACKGROUND: The incidence of hepatocellular carcinoma (HCC) is very high in the world. However, a safe and effective strategy is still under research. AIMS: Our aim was to demonstrate the inhibitory effect of Shaoyao Ruangan Formmula (SRF) on the tumor of H22-bearing mice and explore its antitumor mechanisms. SETTINGS AND DESIGN: Corresponding physiological indexes of H22-bearing mice treated with SRF were compared with that of saline treated mice, which could reflect the tumor-suppressing effect of SRF. MATERIALS AND METHODS: After treatment, tumor weight, survival time, related gene expression levels etc., were recorded or detected. STATISTICAL ANALYSIS: Data analyzed using a computer SPSS program. RESULTS AND CONCLUSIONS: Comparing with blank control group, the tumor inhibitor rate (IR) of low, middle and high dose group of SRF was 17.72%, 33.99% and 23.73%, respectively. IR of CTX was 43.95%. The results also showed that each group of SRF could prolong the life span of H22-bearing mice to some extent. In addition, reverse transcription polymerase chain reaction (RT-PCR) results revealed that SRF was able to influence related genes expression in the tumor tissues of H22-bearing mice. The expression of TGF-ß receptor type II (TBRII) gene was significantly upregulated in each SRF group comparing with normal saline group. On the contrary, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) was significantly downregulated in each SRF group comparing with normal saline group. In summary, SRF showed tumor-suppressing effect on mice with transplanted H22 hepatocarcinoma. The mechanism of antitumor effect may induced by upregulating TBRII expression and down-regulating NF-κB expression.

[HPLC-fingerprint-based quality evaluation on a Tibetan medicine Phyllanthus emblica and its tannin parts].

This study is to establish the fingerprint for Phyllanthus emblica and their tannin parts from different habitats by HPLC for its quality control. The determination was carried out on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column, with methanol-0.2% glacial acetic acid as mobile phase with gradient elution at a flow rate of 1 mL x min(-1). The temperature was maintained at 30 degrees C and the detected wavelength is 260 nm, Thirteen chromatographic peaks were extracted as the common peaks of the fingerprint of P. emblica, and eleven as the common peaks of P. emblica tannin parts, and five peaks were identified by comparing with referent samples. The fingerprints of 8 samples were compared and classified by similarity evaluation, cluster analysis and principal component analysis (PCA). The similarity degrees of eight P. emblica were between 0.763 and 0.993, while tannin parts were between 0.903 and 0.991. All the samples of P. emblica and their tannin parts were classified into 3 categories. The method was so highly reproducible, simple and reliable that it could provide basis for quality control and evaluation of P. emblica from different habitats.

Extracts from Curcuma zedoaria Inhibit Proliferation of Human Breast Cancer Cell MDA-MB-231 In Vitro.

Objective. To evaluate the effect of petroleum ether extracts of Curcuma zedoaria on the proliferation of human triple negative breast cancer cell line MDA-MB-231. Methods. The reagents were isolated from Curcuma zedoaria by petroleum ether fraction. It was assayed by CCK8 for MDA-MB-231 cellular viability with various concentrations and days, cell cycle analyses, Western Blot analysis, and Realtime Reverse Transcriptase PCR analyses for chemokines molecules including E-cadherin, and E-selectin, and adhesion molecules including CCR7, SLC, SDF-1, and CXCR4. Epirubicin was used as control in the study. Results. MDA-MB-231 cells were inhibited by petroleum ether extracts of Curcuma zedoaria (P < 0.05), and the inhibition rate was dependent on concentrations and time. Petroleum ether extracts of Curcuma zedoaria as well as Epirubicin produce a significant G0/G1 cell cycle arrest. The level of expression of proteins E-cadherin and E-cadherin mRNA was significantly increased, while proteins SDF-1, CCR7, and CXCR4 mRNA were decreased after being incubated with petroleum ether extracts of Curcuma zedoaria at the concentrations of 300 µ g/mL than control (P < 0.05). The differences were that the protein CXCR4 mRNA expression level was higher than vehicle. Conclusions. MDA-MB-231 cells were inhibited by petroleum ether extracts of Curcuma zedoaria.

[Effects of actinomycetes agent on ginseng growth and rhizosphere soil microflora].

Taking the ginseng in Xiao Xing' an Mountains of Northeast China as test object, this paper studied the effects of applying Streptomyces pactum (Act12) on ginseng growth and on the soil microflora in root zone and root surface. After treated with Act12, the yield and quality of ginseng' s medicinal part improved, the induced enzyme activities in leaves and the root activity increased, and the numbers and proportions of soil bacteria and actinomycetes increased significantly while those of soil fungi decreased. Compared with the control, the soil microflora in treatment Act12 changed. The numbers of the dominant bacteria Pseudomonas fluorescens, Pseudomonas koreensis, and Microbacterium oxydans were much higher in root zone soil and root surface soil, and the pathogen Plectosphaerella cucumerina decreased in root zone soil and disappeared in root surface soil. These results suggested that the addition of Act12 could improve the soil microflora, enhance the resistance and root activity of ginseng plant, and increase the ginseng yield and its quality.

Tupisteroide A-C, three new polyhydroxylated steroidal constituents from the roots of Tupistra chinensis.

Three new steroidal compounds with polyhydroxy groups, tupisteroide A-C (1-3), were obtained from the roots of Tupistra chinensis, together with one known compound (4) that was isolated from this plant for the first time. The structures of tupisteroide A-C were determined on the basis of one- and two-dimensional NMR spectroscopy, including (1) H-(1) H Correlation Spectroscopy, Heteronuclear Multiple Bond Correlation, and Heteronuclear Single Quantum Coherence experiments. The isolated compounds were evaluated for their cytotoxic activities against A549, HepG2, and CaSki cancer cell lines in vitro. Among them, compounds 1, 2, and 4 did not show significant inhibitory activity, but compound 3 showed cytotoxicity against A549 cancer cell lines with IC(50) values of 25.0 µM.

[Study on spaceflight mutagenesis Platycodon grandiflorum via FTIR].

OBJECTIVE: To establish a method for analyzing amorphous organic compound components of space Platycodon grandflorum rapidly. METHOD: FTIR was applied for measuring and comparing P. grandflorum of comparison group,the ground group and the 4" generation of space group. RESULT: Several active components (platycodin, polysaccharide etc.) contents of space group are increased obviously. CONCLUSION: The major components and the structures remained inextenso,and the effictive component contents are enhanced in the space P. grandiflorum. FTIR is a fast method to analyze the changes of amorphous organic compound components of the space traditional Chinese medicines.

[Effects of different enteral nutrition formulae combined with recombinant human growth hormone on inflammatory response in rats with scald injury].

OBJECTIVE: To investigate the effects of enteral immunonutrition or enteral nutrition combined with recombinant human growth hormone (rhGH) on inflammatory response in rats with severe scald. METHODS: One hundred and twenty-eight SD rats were inflicted with 30% TBSA full-thickness scald, then they were divided into enteral nutrition (EN), enteral immunonutrition (EIN), EN combined with rhGH (EN + rhGH) and EIN combined with rhGH (EIN + rhGH) groups according to random number table. The serum levels of endotoxin, IL-6 and TNF-alpha as well as the expression of CD14 and TNF-alpha mRNA in liver were determined on the 1, 4, 7, 10 post scald day (PSD). Eight normal SD rats served as control, and above indices were also examined in this group (C group). RESULTS: The levels of endotoxin, IL-6 and TNF-alpha as well as the expression of CD14mRNA, TNF-alphamRNA in EN, EIN, EN + rhGH, EIN + rhGH groups were remarkably higher than those of C group at each time point. The levels of endotoxin, IL-6 and TNF-alpha as well as the expression of CD14 mRNA, TNF-alpha mRNA in EIN and EN + rhGH groups were lowered significantly on 4, 7 and 10 PSD compared with those of EN group (P < 0.05 or P < 0.01). The levels of endotoxin (0.37 +/- 0.07 EU/mL), IL-6 (289 +/- 49 ng/L) and TNF-alpha (1.87 +/- 0.32 microg/L) as well as the expression of CD14 mRNA (0.39 +/- 0.05), TNF-alpha mRNA (0.47 +/- 0.03) in EIN + rhGH group were also obviously lower than those [(0.48 +/- 0.08) EU/mL, (364 +/- 53) ng/L, (2.50 +/- 0.48) microg/L, 0.67 +/- 0.06, 0.66 +/- 0.05, P < 0.05 or P < 0.01, respectively] in EIN group on 10 PSD (P < 0.05 or P < 0.01). CONCLUSION: Enteral immunonutrition combined with rhGH can ameliorate inflammatory response, in which rhGH has a collaborative effect.

[Effect of acupuncture-moxibustion combined with nerve growth factor on compensation of cerebral function in the children of cerebral palsy].

OBJECTIVE: To investigate the compensation of cerebral function in acupuncture for rehabilitation of cerebral palsy. METHODS: One hundred children of cerebral palsy were randomly divided into a treatment group and a control group, 50 cases in each group. The treatment group were treated with scalp acupuncture at the Motor Area, Foot Motor Sensory Area and Equilibrium Area, body acupuncture at Binao (LI 14), Fengchi (GB 20), Huantiao (GB 30), etc. and injection of nerve growth factor into Zusanli (ST 36), in combination with rehabilitation training; the control group were treated only with rehabilitation training. Their clinical therapeutic effects and recoveries of brain lesion detected by CT, SEPCT were investigated. RESULTS: The total effective rate was 84.0% in the treatment group better than 52.0% of the control group. After treatment, the development quotient (DQ) in the treatment group was higher than the control group (P<0.01). CONCLUSION: Acupuncture can promote compensation of cerebral function in the children of cerebral palsy.

[Impact of Chai Qin Cheng Qi Decoction on cholinergic anti-inflammatory pathway in rats with severe acute pancreatitis].

OBJECTIVE: To test the change of true choline esterase (TChE) and cholineacetyltransferase (ChAT) and their correlation with interleukin-6 (IL-6) and Tumor Necrosis Factor-alpha (TNF-alpha), and the impact of Chai Qin Cheng Qi Decoction (CQCQD) on cholinergic anti-inflammatory pathway in rats with severe acute pancreatitis (SAP). Methods Thirty SD rats were randomly divided into three groups: control group (pseudo-operated), SAP group and CQCQD-treated group, each with 10 rats. Blood samples were taken six hours after injection of testing agents for biochemical test, which included the test of amylase, TNF-alpha, IL-6, TChE, and ChAT. RESULTS: The rats in SAP group had higher levels of serum IL-6, TNF-alpha and TChE and lower levels of serum ChAT than those in control group (P < 0.05). The serum IL-6 was positively correlated with TChE (r = 0.95, P = 0.000) and negatively correlated with ChAT (r = -0.91, P = 0.000). The TNF-alpha was also positively correlated with TChE (r = 0.93, P = 0.000) and negatively correlated with ChAT (r = -0.95, P = 0.004). The rats in CQCQD-treated group had lower levels of serum IL-6, TNF-alpha and TChE and higher levels of serum ChAT than those in SAP group (P < 0.01). The increase of white blood cell, alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase was observed in the control group first and followed by the CQCQD-treated group and SAP group sequentially (P < 0.05). Conclusions Cholinergic anti-inflammatory pathway plays an important role in the pathological changes of SAP in rats. CQCQD can relieve the systemic inflammatory response syndrome and reduce the functional damage of organs through interference on the cholinergic antiinflammatory pathway. More studies are needed to reveal the mechanism of such impact.

[Clinical study on severe acute pancreatitis complicated by hepatic insufficiency].

OBJECTIVE: To investigate the pathogenic factors of severe acute pancreatitis (SAP) complicated by hepatic insufficiency, the prognosis and the effective preventive and therapeutic interventions. METHODS: One hundred and fifty-two patients with SAP (from January 2003 to June 2004) were divided into 2 groups, SAP with hepatic insufficiency group and SAP without hepatic insufficiency group. The related factors such as causes of disease, serum biochemical criteria, complications, mortality and course of disease were observed. RESULTS: The concentrations of serum amylase, creatinine and lactate dehydrogenase were all much higher in the SAP with hepatic insufficiency group than those in the SAP without hepatic insufficiency group (P<0.05). The incidences of complications such as acute renal failure, heart failure, gastrointestinal hemorrhage and infection were also higher in the SAP with hepatic insufficiency group than those in the SAP without hepatic insufficiency group (P<0.05). There was no significant difference in mortality between these two groups, but the course of disease in SAP with hepatic insufficiency group was longer than that in the other group (P<0.05). CONCLUSION: The causes of SAP complicated by hepatic insufficiency are related to the imbalance of internal environment and the characteristics of the liver function. To control the biliary tract diseases, intervene with traditional Chinese medicine, recover the intestinal function in time, reduce the damage of pancreatic enzyme, maintain the stability of internal environment and avoid using drugs that can induce liver injury are all important aspects of reducing the incidence of hepatic insufficiency.