Acupoint transplantation versus non-acupoint transplantation using autologous peripheral blood mononuclear cells in treating peripheral arterial disease.

Numerous studies have discussed the therapeutic outcomes of using cell therapy or acupuncture to treat peripheral artery disease (PAD). However, there are no long-term studies on the safety and efficacy of transplanting peripheral blood mononuclear cells (PBMNCs) via acupoints to treat PAD. We first reviewed the short-term and long-term clinical results of PAD patients treated with PBMNCs through intramuscular non-acupoint transplantation (control group; n = 45) or intramuscular acupoint transplantation (acupoint group; n = 45) at a single university hospital general medical center between December 2002 and September 2022. Pain intensity (assessed with the verbal rating scale [VRS] score) in the acupoint group was considerably lower than that in the control group at month 1 (mean ± standard deviation [SD]: 1.29 ± 0.96 vs 1.76 ± 0.82; P = 0.016) and month 3 (mean ± SD: 1.27 ± 0.90 vs 1.61 ± 0.86; P = 0.042). We observed significant improvement of VRS score (P < .001 for all) and ankle-brachial index (ABI; P < .001 for all) from baseline in both groups at months 1, 3, 6, 12, 36, and 60. The 10-year cumulative rate of major amputation-free survival (MAFS) was higher in the acupoint group as compared to the control group (81.9%, 95% confidence interval [CI]: 71.3%-94.1% vs 78.5%, 95% CI: 66.7%-92.3%; P = 0.768). Compared with the routine injection method, intramuscular transplantation of PBMNCs via selected acupoints could significantly decrease the short-term pain intensity in patients with PAD, which remains an option for consideration.

Implications on growth performance, glucose metabolism, PI3K/AKT pathway, intestinal flora induced by dietary taurine in a high-carbohydrate diet for grass carp (Ctenopharyngodon idella).

An 8-week experiment was performed to investigate the influence on growth performance, plasma biochemistry, glucose metabolism and the insulin pathway of supplementation of dietary taurine to a high-carbohydrate diet for grass carp. In this study, fish were fed diets at one of two carbohydrate levels, 31·49 % (positive control) or 38·61 % (T00). The high-carbohydrate basal diet (T00), without taurine, was supplemented with 0·05 % (T05), 0·10 % (T10), 0·15 % (T15) or 0·20 % (T20) taurine, resulting in six isonitrogenous (30·37 %) and isolipidic (2·37 %) experimental diets. The experimental results showed that optimal taurine level improved significantly weight gain, specific growth rate (SGR), feed utilisation, reduced plasma total cholesterol levels, TAG and promoted insulin-like growth factor level. Glucokinase, pyruvate kinase and phosphoenolpyruvate carboxykinase activities showed a quadratic function model with increasing dietary taurine level, while hexokinase, fatty acid synthetase activities exhibited a positive linear trend. Optimal taurine supplementation in high-carbohydrate diet upregulated insulin receptor (Ir), insulin receptor substrate (Irs1), phosphatidylinositol 3-kinase (pi3k), protein kinase B (akt1), glycogen synthase kinase 3 ß (gs3kß) mRNA level and downregulated insulin-like growth factor (igf-1), insulin-like growth factor 1 receptor (igf-1R) and Fork head transcription factor 1 (foxo1) mRNA level. The above results suggested that optimal taurine level could improve growth performance, hepatic capacity for glycolipid metabolism and insulin sensitivity, thus enhancing the utilisation of carbohydrates in grass carp. Based on SGR, dietary optimal tributyrin taurine supplementation in grass carp was estimated to be 0·08 %.

Effect of ultrasound-assisted osmotic dehydration on the drying kinetics, water state, and physicochemical properties of microwave vacuum-dried potato slices.

The effects of pre-treatments on the drying characteristics, water state, thermal properties, and bulk shrinkage of potato slices during microwave vacuum drying (MVD) were investigated. The pre-treatment included ultrasound in distilled water (USOD-0%), and ultrasound-assisted osmotic dehydration in a 60% sucrose solution (USOD-60%). Results showed that the drying time of potato slices was reduced and the drying rate was increased when USOD-0% was used as a pretreatment, whereas USOD-60% had a negative effect on the drying rate of the samples. The Weibull model was effective in predicting the water changes in potato slices during the drying process. NMR analysis revealed that the relative content of immovable water (M22) increased initially, then decreased for drying, while the transverse relaxation time (T2) and the relative content of free water (M23) decreased consistently. The DSC results indicated that the glass transition temperature (Tg) had an inverse relationship with the water content of the samples, yet had virtually no influence on the volume shrinkage. The sample volumes decreased linearly with the decrease in water content during the initial drying stages. USOD pre-treatment lessened the volume shrinkage of MVD potato slices. Static gravimetry was used to determine the moisture sorption isotherms of MVD potato slices at 30 °C within the aw range of 0.113-0.923. The GAB model accurately fitted the experimental sorption data, which showed sigmoid shape curves for the MVD samples. When aw values exceeded 0.7, the USOD-60% treatment significantly reduce the water sorption capacity of MVD potato slices, while USOD-0% treatment was observed to increase the hygroscopic properties of MVD samples. Compared with the control, USOD-0% pretreatment significantly increased the monolayer water content (X0), sorption surface area (S0), the thickness of sorbed water multilayer (tm), and density of sorbed water (Ds) values of MVD potato slices, while USOD-60% decreased these values.

Effect of transdermal drug delivery therapy on anxiety symptoms in schizophrenic patients.

Objective: To evaluate the efficacy and safety of transdermal drug delivery therapy for schizophrenia with anxiety symptoms. Methods: A total of 80 schizophrenic patients (34 males and 56 females) with comorbid anxiety disorders were randomly assigned to the treatment group (n = 40) and the control group (n = 40) with 6 weeks of follow-up. The patients in the treatment group received the standard antipsychotic drug treatment along with transdermal drug delivery therapy. The evaluation of the patients included the Hamilton Anxiety Scale (HAMA), Hamilton Depression Scale (HAMD-17), and treatment emergent symptom scale (TESS) at baseline, 3 weeks, and 6 weeks after transdermal drug delivery therapy. The Positive and Negative Symptom Scale (PANSS) was assessed at baseline and after 6 weeks of treatment. Results: After 3 and 6 weeks of treatment, the HAMA scale scores in the treatment group were lower than those in the control group (p < 0.001). However, there were no significant differences in the HAMD-17 scale scores, PANSS total scores, and subscale scores between the two groups (p > 0.05). Additionally, no significant differences in adverse effects were observed between the two groups during the intervention period (p > 0.05). After 6 weeks of penetration therapy, there was a low negative correlation between total disease duration and the change in HAMA scale score (pretreatment-posttreatment) in the treatment group. Conclusion: Combined traditional Chinese medicine directed penetration therapy can improve the anxiety symptoms of patients with schizophrenia and has a safe profile.

[Effect of acupotomy on the fat infiltration degree of lumbar multifidus muscle in patients with lumbar disc herniation after percutaneous transforaminal endoscopic discectomy].

OBJECTIVE: To observe the effect of acupotomy on the fat infiltration degree of lumbar multifidus muscle (LMM) in patients with lumbar disc herniation after percutaneous transforaminal endoscopic discectomy (PTED). METHODS: A total of 104 patients with lumbar disc herniation treated with PTED were randomly divided into an observation group (52 cases, 3 cases dropped off) and a control group (52 cases, 4 cases dropped off). Patients of both groups received rehabilitation training of two weeks 48 h after PTED treatment. The observation group was treated with acupotomy (L3-L5 Jiaji [EX-B 2]) once within 24 h after PTED. In the two groups, the fat infiltration cross sectional area (CSA) of LMM was compared before and 6 months after PTED, the visual analogue scale (VAS) score and Oswestry disability index (ODI) score were observed before and 1, 6 months after PTED. The correlation between fat infiltration CSA of LMM in each segment and VAS score was analyzed. RESULTS: Six months after PTED, the fat infiltration CSA of LMM in L4/L5 and the total L3-S1 segments of the observation group was lower than that before PTED (P<0.05), and the fat infiltration CSA of LMM in L4/L5 of the observation group was lower than the control group (P<0.01). One month after PTED, the ODI and VAS scores of the two groups were lower than those before PTED (P<0.01), and those in the observation group were lower than the control group (P<0.05). Six months after PTED, the ODI and VAS scores of the two groups were lower than those before PTED and 1 month after PTED (P<0.01), and those in the observation group were lower than the control group (P<0.01). There was a positive correlation between the fat infiltration CSA of LMM in the total L3-S1 segments and VAS scores in the two groups before PTED (r = 0.64, P<0.01). Six months after PTED, there was no correlation between the fat infiltration CSA of LMM in each segment and VAS scores in the two groups (P>0.05). CONCLUSION: Acupotomy can improve the fat infiltration degree of LMM, pain symptoms and activities of daily living in patients with lumbar disc herniation after PTED.

[Simultaneous determination of 11 neurotransmitters in brain microdialysis samples from rats by UPLC-MS/MS].

This study established a method for simultaneous determination of 11 neurotransmitters, such as acetylcholine, glutamic acid, glycine, and norepinephrine from rat brain microdialysis samples using UPLC-MS/MS. A total of 20 µL of rat brain dialysate was diluted with 60 µL of acetonitrile-water(4∶1) and centrifuged for 10 min at 10 000 r·min~(-1),and 5 µL was injected into UPLC-MS/MS system for assay. Chromatographic separation was performed on a Waters ACQUITY BEH amide column(2.1 mm×100 mm, 1.7 µm) with gradient elution using acetonitrile/0.2% formic acid-water as mobile phases with a flow rate of 0.35 mL·min~(-1) and column temperature of 35 ℃. The eluate was detected by multiple-reaction monitoring(MRM) scanning with an electrospray ionization source operating in the positive ionization mode with an analysis duration of 3.5 min. The relationship between the recovery rate of 11 neurotransmitters and the perfusion rate or the concentration of neurotransmitters was investigated. Furthermore, the effects of puerarin alone or combined with borneol on the content of 11 neurotransmitters in the striatum of rats were investigated. The results showed the calibration curves displayed good linear regression with coefficients all greater than 0.99 and the lower limit of quantification(LLOQ) less than 12.5 nmol·L~(-1) for each analyte. The within-run and between-run precision(RSD) of the 11 neurotransmitters at low, medium, and high levels was less than 9.3%, and the relative error of the accuracy ranged from-8.4% to 9.5%. The stability, recovery, and matrix effects were in line with the biological sample analysis requirements. As revealed by experimental results, the levels of most neurotransmitters in the brain striatum changed significantly after rats were treated with puerarin as compared with the conditions in the blank group. Except for dopamine and norepinephrine, the degree of changes of other neurotransmitters in the combination group(borneol and puerarin) was less than that of the puerarin group. The established UPLC-MS/MS method could be applied to the quantitative determination of 11 neurotransmitters in microdialysis samples, providing an efficient and useful tool to study neurotransmitter changes in animal models of health and diseases.

Integrating network pharmacology and experimental models to investigate the efficacy of QYHJ on pancreatic cancer.

ETHNOPHARMACOLOGICAL RELEVANCE: The Qingyihuaji decoction (QYHJ) is composed of seven herbs: Scutellaria barbata D.Don (Banzhilian, HSB), Gynostemma pentaphyllum (Thunb.) Makino (Jiaogulan, GP), Oldenlandia diffusa (Willd.) Roxb. (Baihuasheshecao, HDH), Ganoderma lucidum (Leyss. ex Fr.) Karst. (Lingzhi, GL), Myristica fragrans Houtt. (Doukou, AK), and Amorphophallus kiusianus (Makino) Makino (Sheliugu, RA), and Coix lacryma-jobi var. ma-yuen (Rom.Caill.) Stapf (Yiyiren, CL). QYHJ has been reported to exhibit clinical efficacy in the treatment of pancreatic adenocarcinoma (PAAD). However, the molecular mechanism remains unclear. AIM OF THE STUDY: This study explores the therapeutic mechanism of QYHJ in the treatment of PAAD using network pharmacology to identify related targets and pathways in vivo and in vitro. MATERIALS AND METHODS: The bioactive compounds of QYHJ were retrieved and screened using the ADME network pharmacology approach, followed by compound-target prediction and overlapping genes between PAAD oncogenes and QYHJ target genes. The compound-target-pathway network was established using The KEGG pathway, GO analysis, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) analysis to identify potential action pathways. The effects of QYHJ on PAAD were evaluated in vivo and in vitro, and the predicted targets and potential pathways related to QYHJ in PAAD treatment were evaluated using qRT-PCR and immunoblotting. RESULTS: A total of 68 bioactive compounds of QYHJ fulfilled the ADME screening criterion, and their respective 242 target genes were retrieved. The compound-target-disease network identified 11 possible target genes. The KEGG pathway analysis showed significant enrichment of pathways in cancers, involving regulating cancer-related pathways of inflammation, oxidative stress, and apoptosis. Furthermore, QYHJ inhibited PAAD growth in vivo; suppressed cell proliferation, invasion, and migration of PAAD; and induced cellular apoptosis in vitro. The qRT-PCR results showed that QYHJ suppressed the mRNA expression of ICAM1, VCAM1, and Bcl2, and increased that of HMOX1 and NQO1. Immunoblotting revealed changes in the PI3K/AKT/mTOR, Keap1/Nrf2/HO-1/NQO1, and Bcl2/Bax pathways upon QYHJ treatment. CONCLUSIONS: QYHJ can suppress PAAD growth and progression through various mechanisms, including anti-inflammation and apoptosis-induction.

Effects of the Clostridium butyricum on growth performance, antioxidant capacity, immunity and disease resistance of Litopenaeus Vannamei fed with cottonseed protein concentrate (CPC) replacement of fishmeal in diet.

Clostridium butyricum (CB) is a gram-positive bacterium that secretes short-chain fatty acids such as butyric acid and so on. An 8-week feeding trial was conducted to investigate the effects of CB on the growth performance, antioxidant capacity, immunity and resistance to Vibrio parahaemolyticus in Litopenaeus Vannamei fed with cottonseed protein concentrate (CPC) replacement of fishmeal. Six iso-nitrogenous (40%) and iso-lipidic (6%) diets were formulated including a positive control group (PC, 25% fishmeal), a negative control group (NC, CPC replaced 30% of fishmeal protein), and 0.03% (C1, 3 × 108 CFU/kg), 0.12% (C2, 1.2 × 109 CFU/kg), 0.48% (C3, 4.8 × 109 CFU/kg) and 1.92% (C4, 1.92 × 1010 CFU/kg) CB were supplemented on the negative control group (NC). After the feeding trial, the remaining shrimp in each treatment group were subjected to a challenge experiment with Vibrio parahaemolyticus. The results indicated that weight gain rate (WGR), specific growth rate (SGR) in C4 group were significantly lower than those in PC and C2 groups (P < 0.05); the feed conversion ratio (FCR) was significantly higher than that of PC and C2 groups (P < 0.05). There was no significant difference in survival rate (SR) among all groups (P > 0.05). Compared to the PC and NC groups, the total superoxide capacity, superoxide dismutase and lysozyme were significantly higher in the C4 group (P < 0.05); the glutathione peroxidase, acid phosphatase and alkaline phosphatase were significantly higher in the C3 group (P < 0.05); and the malondialdehyde was significantly lower in the C4 group (P < 0.05). The relative mRNA expressions of Toll receptor (TLR), innate immune deficiency gene (IMD), penaiedin3a (Pen3) were significantly down-regulated in the NC group than those in the PC group (P < 0.05). In addition, the relative mRNA expressions of TLR, IMD and Pen3 were significantly up-regulated in all groups supplemented with CB than those in the NC group (P < 0.05). Moreover, the cumulative mortality rate in the NC group was not significantly different from the PC group (P > 0.05) and was significantly higher than those in the C3 and C4 groups (P < 0.05). In conclusion, the CB supplementation on the basis of CPC replacement of 30% fishmeal protein enhanced significantly the antioxidant capacity, immunity and disease resistance of shrimp and improved its growth performance. Therefore, considering the factors of the growth, immunity and disease resistance, the CB supplementation of 0.12%-0.48% (1.2 × 109 CFU/kg-4.8 × 109 CFU/kg) was recommended in the diet of L. vannamei based on the results of this experiment.

[Ginsenoside Rg_1 protects PC12 cells against Aß-induced injury through promotion of mitophagy by PINK1/parkin activation].

Amyloid ß-protein(Aß) deposition in the brain is directly responsible for neuronal mitochondrial damage of Alzheimer's disease(AD) patients. Mitophagy, which removes damaged mitochondria, is a vital mode of neuron protection. Ginsenoside Rg_1(Rg_1), with neuroprotective effect, has displayed promising potential for AD treatment. However, the mechanism underlying the neuroprotective effect of Rg_1 has not been fully elucidated. The present study investigated the effects of ginsenoside Rg_(1 )on the autophagy of PC12 cells injured by Aß_(25-35) to gain insight into the neuroprotective mechanism of Rg_1. The autophagy inducer rapamycin and the autophagy inhi-bitor chloroquine were used to verify the correlation between the neuroprotective effect of Rg_1 and autophagy. The results showed that Rg_1 enhanced the viability and increased the mitochondrial membrane potential of Aß-injured PC12 cells, while these changes were blocked by chloroquine. Furthermore, Rg_(1 )treatment increased the LC3Ⅱ/Ⅰ protein ratio, promoted the depletion of p62 protein, up-regulated the protein levels of PINK1 and parkin, and reduced the amount of autophagy adaptor OPTN, which indicated the enhancement of autophagy. After the silencing of PINK1, a key regulatory site of mitophagy, Rg_1 could not increase the expression of PINK1 and parkin or the amount of NDP52, whereas it can still increase the LC3Ⅱ/Ⅰ protein ratio and promote the depletion of OPTN protein which indicated the enhancement of autophagy. Collectively, the results of this study imply that Rg_1 can promote autophagy of PC12 cells injured by Aß, and may reduce Aß-induced mitochondrial damage by promoting PINK1-dependent mitophagy, which may be one of the key mechanisms of its neuroprotective effect.

[Clinical observation on Tongdu Tiaoqi acupuncture combined with warming acupuncture for postoperative urinary retention].

OBJECTIVE: To compare the therapeutic effect of Tongdu Tiaoqi acupuncture (acupuncture for unblocking governor vessel and regulating qi ) combined with warming acupuncture, Tongdu Tiaoqi acupuncture, abdominal moxibustion and oral tamsulosin hydrochloride sustained release capsule on postoperative urinary retention. METHODS: A total of 120 patients with postoperative urinary retention were randomized into an acupuncture-moxibustion group, an acupuncture group, a moxibustion group and a medication group, 30 cases in each group. Tongdu Tiaoqi acupuncture combined with warming acupuncture were applied in the acupuncture-moxibustion group. Tongdu Tiaoqi acupuncture was applied at Baihui (GV 20), Shuigou (CV 26) etc. in the acupuncture group. Moxibustion was applied at Qihai (CV 6), Guanyuan (CV 4), Shuidao (ST 28) and Sanyinjiao (SP 6) in the moxibustion group. Tamsulosin hydrochloride sustained release capsule was given orally in the medication group. The treatment was once a day, and 5-day treatment was required in each group. Before and after treatment, the residual urine volume of bladder, the visual analogue scale (VAS) score and the time of first urethral catheter removal were observed, and the clinical efficacy was compared in the 4 groups. RESULTS: After treatment, the residual urine volume of bladder was decreased compared before treatment in the 4 groups (P<0.05), and that in the acupuncture-moxibustion group was less than the other 3 groups (P<0.05). After treatment, the VAS scores were decreased compared before treatment in the acupuncture-moxibustion group, the acupuncture group and the moxibustion group (P<0.05), and those in the 3 groups were lower than the medication group (P<0.05). The time of first urethral catheter removal in the acupuncture-moxibustion group was earlier than the other 3 groups (P<0.05). The total effective rate was 93.3% (28/30) in the acupuncture-moxibustion group, which was superior to 63.3% (19/30) in the acupuncture group, 60.0% (18/30) in the moxibustion group and 66.7% (20/30) in the medication group (P<0.05). CONCLUSION: The therapeutic effect of Tongdu Tiaoqi acupuncture combined with warming acupuncture on postoperative urinary retention is superior to simple acupuncture, abdominal moxibustion and tamsulosin hydrochloride sustained release capsule.

Matrine injection inhibits pancreatic cancer growth via modulating carbonic anhydrases- a network pharmacology-based study with in vitro validation.

ETHNOPHARMACOLOGICAL RELEVANCE: Matrine injection is a complex mixture of plant bioactive substances extracted from Sophora flavescens Aiton and Smilax glabra Roxb. Since its approval by the Chinese Food and Drug Administration (CFDA) in 1995, Matrine injection has been clinically used as a complementary and alternative treatment for various cancers; however, the underlying mechanism of pancreatic cancer treatment is yet to be elucidated. AIM OF THE STUDY: The present study explores the potential mechanism of matrine injection on pancreatic cancer through network pharmacology technique and in vitro experimental validation. MATERIALS AND METHODS: Genes differentially expressed in pancreatic cancer were obtained from the Gene Expression Omnibus (GEO) database (GSE101448). The potential active components of matrine injection were selected following a literature search, and target prediction was performed by the SwissTarget Prediction database. Overlapping genes associated with survival were screened by the Gene Expression Profiling Interactive Analysis (GEPIA) database. In vitro experimental validation was performed with cell counting kit-8 (CCK-8) assay, apoptosis detection, cell cycle analysis, immunoblotting, and co-immunoprecipitation of the identified proteins. RESULTS: One thousand seven hundred genes differentially expressed among pancreatic tumor and non-tumor tissues were screened out. Sixteen active components and 226 predicted target genes were identified in matrine injection. A total of 25 potential target genes of matrine injection for the treatment of pancreatic cancer were obtained. Among them, the prognostic target genes carbonic anhydrase 9 (CA9) and carbonic anhydrase 12 (CA12) based on the GEPIA database are differently expressed in tumors compared to adjacent normal tissue. In vitro experiments, the results of CCK-8 assay, apoptosis and cell cycle analysis, immunoblotting, and co-immunoprecipitation showed that matrine injection inhibited Capan-1 and Mia paca-2 proliferation, arrested the cell cycle at the S phase, and induced apoptosis through up-regulated CA12 and down-regulated CA9. CONCLUSIONS: In this study, bioinformatics and network pharmacology were applied to explore the treatment mechanism on pancreatic cancer with matrine injection. This study demonstrated that matrine injection inhibited proliferation, arrested the cell cycle, and induced apoptosis of pancreatic cancer cells. The mechanism may be related to the induction of CA12 over-expression, and CA9 reduced expression. As novel targets for pancreatic cancer treatment, Carbonic anhydrases require further study.

Ginkgo biloba Extract EGb761 Attenuates Bleomycin-Induced Experimental Pulmonary Fibrosis in Mice by Regulating the Balance of M1/M2 Macrophages and Nuclear Factor Kappa B (NF-κB)-Mediated Cellular Apoptosis.

BACKGROUND The aim of this study was to show whether the standardized Ginkgo biloba extract EGb761, a traditional Chinese medicine, has a therapeutic effect on pulmonary fibrosis (PF). MATERIAL AND METHODS Bleomycin (BLM) was used for establishing the PF mouse model. The mice were treated with a gradient of EGb761 for 28 days to determine an appropriate drug dose. On day 28, the effect of EGb761 on lung injury and inflammation was confirmed by hematoxylin and eosin and Masson staining and evaluated by pulmonary alveolitis and Ashcroft score. The balance of M1/M2 macrophages was evaluated with the respective markers inducible nitric oxide synthase and and interleukin-10 by real-time polymerase chain reaction. Furthermore, the expressions of fibrosis-associated protein alpha-smooth muscle actin (SMA), related inflammatory protein transforming growth factor (TGF)-ß1, the apoptosis-related proteins B-cell lymphoma-associated X protein (Bax), B-cell lymphoma (Bcl)-2, caspase-3, caspase-9, and phosphorylated nuclear factor (NF)-kappaB (p65) were assessed by western blot. RESULTS On day 28, PF was induced by treating with BLM, whereas EGb761 suppressed the PF of lung tissue. The BLM-induced imbalance of M1/M2 macrophages was reduced by EGb761. Furthermore, the increasing amounts of alpha-SMA and TGF-ß1 induced by BLM were suppressed by EGb761. In addition, the protein or messenger ribonucleic acid expression levels of phosphorylated NF-kappaB (p65), caspase-3, and caspase-9 were upregulated, whereas Bax and Bcl-2 were downregulated. Treatment with EGb761 restored the levels of these proteins except for caspase-9. CONCLUSIONS This study illustrated the protective effect of EGb761 on BLM-induced PF by regulating the balance of M1/M2 macrophages and NF-kappaB (p65)-mediated apoptosis. The results demonstrated the potential clinical therapeutic effect of EGb761, providing a novel possibility for curing PF.

[Research progress of dummy template molecularly imprinted polymers in separation of natural products].

The dummy template molecularly imprinted polymers not only has such characteristics of normal imprinted polymers as rapid identification, easy preparation, stable structure and multiple reuse, but also can imprint the compounds in natural products that are not suitable as direct template. Therefore, it has drawn more and more attention in the field of the study of natural products. This paper summarizes the methods for the selection of dummy template molecules by investigating the relevant literatures in the past ten years, analyzes the advantages and disadvantages of dummy template molecules in the practical application, and based on the types of natural products active ingredients, this paper is the first to review of the latest progress in extraction and separation of dummy template molecularly imprinted polymers. We believed that this paper could provide references for better applications of the dummy template molecularly imprinted polymers to extract and separate natural products.

Bupleurum polysaccharides ameliorated renal injury in diabetic mice associated with suppression of HMGB1-TLR4 signaling.

Bupleurum polysaccharides (BPs) is isolated from Bupleurum smithii var. parvifolium, a key traditional Chinese medicine. The study was to investigate the effects of BPs on diabetic kidney injury. After two intraperitoneal injections of streptozotozin (STZ) 100 mg·kg-1, renal injury in diabetic mice was induced and BPs was orally administrated at dosages of 30 and 60 mg·kg-1·d-1. The STZ injected mice developed renal function damage, renal inflammation and fibrosis known as diabetic kidney disease (DKD). BPs significantly reduced serum creatinine level and urinary albumin excretion rate, with the attenuated swelling of kidneys. BPs treatment obviously alleviated the pathological damage of renal tissue. The progression of renal injury in BPs treated mice was inhibited with less expression of type IV collagen (Col IV), fibronectin (FN) and α-smooth muscle actin (α-SMA). The inhibition of inflammation in kidney was associated with the reduced level of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). BPs administration suppressed the over-expression of toll like receptor 4 (TLR4) and high-mobility group box 1 (HMGB1) with lowered activity of nuclear factor kappa B (NF-κB) in renal tissue of diabetic mice. Oral administration of BPs effectively prevented the development ofrenal injury in diabetic mice. This study suggested that the protection provided by BPs might affect through the interruption of HMGB1-TLR4 pathway, leading to the inhibition of renal inflammation and fibrotic process.

Pharmacokinetic comparison of five xanthones in rat plasma after oral administration of crude and processed Garcinia hanburyi extracts.

Gamboge, a dried resin secreted by Garcinia hanburyi Hook. f. (Guttiferae), possesses remarkable anticancer activity. However, due to toxicity, it must be processed before use in clinics. Xanthones are the main bioactive ingredients in gamboge. In order to elucidate the influence of processing technology on pharmacological properties of gamboge, an efficient, sensitive, and selective ultra-performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-MS/MS) method of five critical xanthones, including ß-morellic acid (ß-MA), isogambogenic acid (IGNA), gambogenic acid (GNA), R-gambogic acid (GA), and S-GA in rat plasma was established for a comparative pharmacokinetics study of these xanthones after oral administration of crude and processed G. hanburyi extracts. The chromatographic separation of these five xanthones along with an internal standard (I.S.) was carried out on a Waters Acquity UPLC BEH C8 column with a gradient elution method using acetonitrile/0.1% formic acid-water as mobile phases. The eluate was detected by multiple-reaction monitoring (MRM) scanning with an electrospray ionization source operating in the positive ionization mode. Sample preparation involved a liquid-liquid extraction of the five analytes with ethyl acetate. Deoxyschizandrin was employed as an internal standard. This assay method was validated for selectivity, linearity, intra-day and inter-day precision, accuracy, recovery, matrix effect, and stability. The results revealed that the calibration curves displayed good linear regression (r > 0.995), and the lower limit of quantification (LLOQ) was <5.52 ng/mL for each analyte. The intra-day and inter-day precision (RSD) of the five xanthones at low, medium, and high levels was <10.58%, and the bias of the accuracy ranged from -8.54 to 10.2%. All other parameters fulfilled the FDA criteria for bioanalytical validation. In addition, the assay was successfully applied to the determination and pharmacokinetic study of these five xanthones after oral administration of crude and processed gamboge. Furthermore, Cmax of GNA and AUC0-t of IGNA were increased significantly (P < 0.05) after processing, while AUC0-t of ß-MA, R-GA, and S-GA decreased remarkably (P < 0.05), which suggested that processing exerted different effects on the absorption of xanthones. The results might be valuable for the clinical reasonable application and understanding the processing mechanism of gamboge.

Mechanism of Yanghe Pingchuan granules treatment for airway remodeling in asthma.

PURPOSE: Yanghe Pingchuan granules (YPG), a hospital preparation developed by The First Affiliated Hospital, Anhui University of Chinese Medicine, has been used for the clinical treatment of bronchial asthma (BA) for several decades. This study aimed to explore the mechanism of action of YPG in the treatment of BA. MATERIALS AND METHODS: Male Sprague Dawley rats (n=60) were randomly divided into six groups (n=10 per group): control, a BA model, positive drug control (Guilong Kechuanning capsules; a proven effective treatment for BA), and model rats treated with a high, medium, or low dose of YPG. H&E staining was used to detect pathological changes in the bronchial tubes. The mRNA expression levels of PI3K, PKB, PCNA, and AR were determined by real-time PCR, and the protein levels of phospho- (p-)PI3K, p-PKB, p-PCNA, and p-AR were detected by Western blotting. ELISAs were used to detect the expression of PIP2, PIP3 IL-6, IL-8, IL-1ß, and epinephrine (EPI). RESULTS: H&E staining demonstrated that BA can be ameliorated using YPG. Real-time PCR, Western blotting, and ELISA indicated that use of YPG decreased expression of the phosphoinositide 3-kinase (PI3K) signaling pathway and PCNA, and can also ameliorate the condition kidney Yang deficiency, which is associated with BA in Chinese traditional medicine. CONCLUSION: YPG can attenuate BA therapeutically in a dose-dependent manner. The mechanism underlying its therapeutic effect comprises influences on three features that contribute to BA: the PI3K signaling pathway, cell proliferation, and "kidney-Yang deficiency".

[Molecular imprinting technology and its application in terpenoids].

Molecular imprinting technology is widely used in the separation and analysis of compounds such as flavonoids, alkaloids and polyphenols, due to its high selectivity and specific recognition and so on. However, no much of attention has been paid to the terpenoids. This paper is aimed to not only review the effects of common synthetic elements such as functional monomers, cross-linking agents and porogens on the polymer properties, but also highlight the application of terpene molecular imprinting in solid phase extraction, sensor, membrane separation and chromatographic separation by means of statistical analysis of literature. Furthermore, the shortcomings and improvement directions are discussed.We believed that this paper could provide references for better applications of molecular imprinting techniques to the analysis of terpenoid compounds.

Six new furostanol glycosides from Smilax glauco-china and their cytotoxic activity.

Six new steroidal saponins, namely glauco-chinaosides A-F, and one known compound were isolated from the tubers of Smilax glauco-china. Their structures were elucidated by a combination of spectroscopic analysis and hydrolysis followed by spectral and chromatographic analysis. Compounds 1-7 were tested in vitro for their cytotoxic activities against four human tumor cell lines (SH-SY5Y, SGC-7901, HCT-116, and Lovo). Compounds 1, 2, and 5 exhibited cytotoxic activity against SGC-7901, with IC50 values of 2.7, 11.5, and 6.8 µM, respectively.

[Effect of Ginkgo biloba Tablet on the Expression of Scavenger Receptor A of the Aortic Wall in Atherosclerotic Rats].

OBJECTIVE: To observe the expression of Ginkgo biloba Tablet (GbT) on scavenger receptor A (SRA) of the aortic wall and changes of serum inflammatory factors in atherosclerotic rats, and to explore its new mechanism for fighting against atherosclerosis (AS). METHODS: Totally 45 male Wistar rats were randomly divided into the control group, the model group, the GbT group, 15 rats in each group. Levels of blood glucose, blood lipids, blood calcium, serum C-reactive protein (CRP), soluble intercellular adhesion molecule-1 (slCAM-1), and soluble vascular cell adhesion molecule-1 (sVCAM-1) were measured in all rats. The expression of SRA in the aortic wall of atherosclerotic rats was observed by immunohistochemical assay. The correlation between the expression of SRA and levels of in-flammatory factors was also observed. RESULTS: Compared with the control group, blood glucose and blood calcium obviously increased (P < 0.05); levels of TG, TC, and LDL-C were significantly elevated (P < 0.01); neointimal areas were significantly thickened, increased intima percentage was significantly enlarged, narrowed lumen index was significantly reduced; levels of CRP, sICAM-1, and sVCAM-1 were significantly elevated in the model group (all P < 0.01). Compared with the model group, blood glucose and blood calcium obviously decreased (P < 0.05); levels of TG, TC, and LDL-C significantly decreased (P < 0.01) in the GbT group. Aortic lumens were obviously narrower in the model group than in the GbT group (P < 0.05). SRA expressed at the aortic wall. The aforesaid 3 indices were significantly improved in the GbT group than in the model group (P < 0.01). Serum levels of CRP, sICAM-1, and sVCAM-1 were significantly decreased in the GbT group than in the model group (P < 0.01). Serum levels of CRP, sICAM-1, and sVCAM-1 were positively correlated with the percentage of SRA positive expression area (r = 0.701, 0.604, 0.581, all P < 0.01). CONCLUSIONS: Serum levels of inflammatory factors in atherosclerotic rats were elevated, and the expression of SRA in the aortic wall was enhanced. The expression of SRA was closely correlated with serum levels of inflammatory factors. GbT could decrease serum levels of inflammatory factors and inhibit the expression of SRA.

[Attenuation mechanism of gamboges during processing based on inflammatory toxicity and AQP3, AQP4 protein and mRNA expressions in rat gastric and duodenal tissues].

To expand the clinical application of gamboges, it is necessary to study crude gamboges' toxicity after oral administration and attenuation mechanism during processing. In this study, crude gamboges' toxicity was judged by multiple assays, including inflammatory mediums [such as nitric oxide (NO), tumor necrosis factor alpha (TNF-α), and interleukin 6 (IL-6)] released by macrophage RAW264.7, and pathological manifestations of rat stomach and duodenal tissues after oral administration with crude and processed gamboges. The attenuation mechanism during processing was studied by detecting AQP3, AQP4 protein and mRNA expression in rat gastric and duodenal tissues using immunohistochemical assay and real-time fluorescent quantitative PCR technique. According to the results, crude gamboges group showed promotion in release of NO, TNF-α and IL-6 by macrophage RAW264.7 in a dose-dependent manner; Compared with crude gamboges group, processed gamboges group showed reduction in release of NO and IL-6, with increase in TNF-α. Crude gamboges could cause rat diarrhea, white blood cells increase, lymphocytes decrease, hyperemia and edema in rat gastric mucosa, duodenal mucosal necrosis and inflammatory cells infiltration. All of these results proved that gamboges had the inflammatory toxicity in gastric and duodenal tissues after oral administration in a dose-dependent manner, which however reduced after processing. In addition to the inflammatory toxicity, the mRNA and protein expressions of aquaporin 3 (AQP3), aquaporin 4 (AQP4) in gastric and duodenal tissues of high-dose crude gamboges group were increased significantly (P<0.05), while the protein and mRNA expressions of AQP3, AQP4 were weakened in processed gamboges group. The results showed that AQP3, AQP4 protein and mRNA expressions were positively correlated with the inflammatory toxicity. In conclusion, down-regulation of AQP3, AQP4 protein and mRNA expressions may be one of attenuation mechanisms in processing gamboges.