Objective To develop a method for simultaneous determination of three hydrophilic components and two lipophilic components in Radix et Rhizoma Salviae Miltiorrhizae. Methods The RP-HPLC method was performed by using a Welchrom C18 column(250 mm×4.6 mm,5 μm)with a mobile phase of acetonitrile (A)-0.1%phosphoric acid(B). The gradient elution program was as follows:0-15 min,10%→12%A;-35 min,12%→20%A;-45 min,20%→60%A;-65 min,60%→65%A;-80 min,65%→80%A;-90 min,10%A. The flow rate was kept at 1.0 mL?min-1 . The detection wavelength was set at 280 nm. The column temperature was 30 ℃ . Results A good linearity was obtained over 0.059 5-2.380 0 μg for tanshinol, 0.346 0-13. 840 0 μg for rosmarinic acid, 0. 656 0 - 26. 240 0 μg for salviamolic acid B, 0. 420 0 - 16.800 0 μg for cryptotanshinone and 0.414 0- 16.560 0 μg for tanshinoneIIA, respectively ( r = 0.999 9). The average recovery rates were between 98.69%-100.91% with RSD less than 1.2%(n = 6). Conclusion The method is rapid, accurate, credible and repeatable, and can provide basis for the quality control of Radix et Rhizoma Salviae Miltiorrhiza.

Objective To explore the efficacy and safety of a raltegravir (RAL)-containing regimen among patients on methadone maintenance therapy.Methods From January 2010 to November 2010, 30 virus (HIV) treatment naive patients who were on methadone maintenance therapy were enrolled from a HIV clinic in Kunming, Yunnan Province and a HIV clinic in Hengyang, Hunan Province.All patients were given RAL, tenofovir (TDF) and lamivudine (3TC) as highly active antiretroviral therapy (HARRT).Patients were followed up for 48 weeks to evaluate the adjustment of methadone dose, opiate withdrawal reaction, antiretroviral efficacy and safety.Results From January 2010 to November 2010, 30 HIV patients were enrolled from the two appointed HIV clinics.The mean age was 39±6 years, with 73.3% male patients and 97% Han population.Before the treatment, their mean CD4+T lymphocyte counts was 210 /μL.Ninety percent of patients were co-infected with hepatitis C.Twenty-nine patients who completed study follow-up were included in final analysis.Five (17.8%) patients reported opiate withdrawal symptoms and increased methadone dose 4 weeks after HARRT.At 24 weeks and 48 weeks of HARRT, the average increase of CD4+T lymphocyte counts were (136±71) /μL and (185±88)/μL, respectively.Among patients who provided valid HIV-1 RNA testing results, 82.6% (19/23) and 95.8% (23/24) of patients had undetectable viral load at week 24 and week 48.Six grade 1-2 adverse events were reported in 4 patients.Conclusions In this pilot study, the new regimen containing RAL, TDF and 3TC appears to be an ideal option for patients on methadone maintenance therapy, because of its limited impact on methadone dose and good efficacy and safety profile.

Objective To study the effects of integrative body-mind training with cognitive rehabilitation based on the sensory integration training theory on executive function and anxiety and depression mood in patients with traumatic brain injury.Methods 85 cases were randomly divided into the control group (42 cases) and the treatment group (43 cases).The control group took targeted cognitive rehabilitation with sensory integration training theory and the treatment group was given targeted integrative bodymind training based on this training.Before training and after 3 months,the cognitive function and executive function were detected by the Behavioral Assessment of Dysexecutive Syndrome (BADS); the cognitive dysfunction were evaluated by Loewenstein Occupation Therapy Cognitive Assessment (LOTCA); Anxiety and Depression were evaluated by Self-rating Depression Scale (SDS) and Self-Rating Anxiety Scale (SAS).Results There was no marked differences in BADS,LOTCA,SDS and SAS scores before treatment; Compared with the control group,they were much improved after training; Moreover,they were much more improved in the treatment group than those of the control group.Conclusions Integrative bodymind training with cognitive rehabilitation based on the sensory integration training theory can improve the executive function and regulate the mood on cognitive dysfunction in patients with traumatic brain injury.

Objective To study the inflammation and expression of myosin light chain kinase(MLCK) through establishing acute lung injury animal model of mice induced by lipopolysacchride(LPS), and approach the role of MLCK in the mechanism of acute lung injury.Methods Twenty female BALB/c mice were randomly divided into LPS group(n=10) and control group(n=10).The BALB/c mice of LPS and control groups were induced by 30 ?L 0.9% NaCl via intranasal instillation,while only LPS group was treated with LPS(20 ?g/each mice).The pathology,wet/dry lung weight ratio and the total cell quantitation in bronchoalveolar lavage fluid(BALF)were compared between these two groups.Furthermore,immunohistochemistry assays were used to determine the status of MLCK expression in the lung.And RT-PCR was adopted to determine the status of MLCKmRNA in the lung. Results Compared with the control group,the LPS group showed more serious pulmonary hemorrhage,edema and infiltration of neutrophils, significantly increased water content in the lungs and total cell quantitation in BALF(P

Objective To study the effect of montelukast(MK)and dexamethasone(Dex)on inflammation of asthma.Methods Asthma model was established and treated with MK or Dex.Bronchoalveolar lavage fluid(BALF) and histopathologic change were observed,IL-5mRNA of lung and bone marrow cells were detected by in situ hybridization,IL-5 immunoreactive cells by immunohistochemistry,and CD34+ and CD3+ of bone marrow cells by flow cytometry. ResultsCompared with asthma group,the number of total cells and eosinophils in BALF of MK group and Dex group were significantly decreased(P0.05). Conclusion MK and Dex can well inhibit airway inflammation and expression of IL-5mRNA in lung and bone marrow cells,though MK may be inferior to Dex in some aspects.The combined treatment of leukotriene receptor antagonist and glucocorticosteroid may be a new direction for asthma.

Objective:To explore the role of group 2 innate lymphoid cells (ILC2) in atopic dermatitis (AD) .Methods:C57BL/6J and Rag1 -/- mice served as research objects. The C57BL/6J mice were divided into 2 groups: model group topically treated with calcipotriol (MC903) on both ears every day for 14 consecutive days, control group topically treated with anhydrous ethanol alone at the same time. On day 15, peripheral blood samples were collected from the mice. After the sacrifice, the ear skin tissues were obtained for histopathological examination, and the spleens were resected. Real-time fluorescence-based quantitative PCR was performed to determine the expression of inflammatory factors in the skin and spleen tissues, and flow cytometry to determine the proportion of ILC2 in the skin tissues. The Rag1 -/- mice were divided into model group, control group and experimental group: the Rag1 -/- mice in the model group and control group received the same treatment and evaluation as the C57BL/6J mice; two days before the topical treatment with MC903, the Rag1 -/- mice in the experimental group started to be intraperitoneally injected with the monoclonal antibody CD90.2 at a dose of 300 μg/150 μl once every other 2 days for 7 sessions, with the purpose of antagonizing the function of ILC2, and other treatments were the same as those in the model group. Skin manifestations were observed, and histopathological features were evaluated. Two-independent-sample t test was used for comparisons between 2 groups, and one-way analysis of variance for comparisons among multiple groups. Results:In the model group, the ear skin of the C57BL/6J mice was apparently red, swollen and dry with crusts, and hematoxylin and eosin (HE) staining showed increased thickness of the epidermis and dermal infiltration of eosinophils; the serum level of IgE (6 751.016 ± 282.324 μg/L) was significantly higher in the model group than in the control group (6 387.038 ± 267.853 μg/L, P= 0.007) , so were the expression of interleukin (IL) -4, IL-13 and interferon (IFN) -γ in the skin tissues ( P= 0.005, 0.012, < 0.001, respectively) , but there was no significant difference in IL-5 expression ( P= 0.190) ; the expression of IL-4, IL-13 and IFN-γ in the spleen was significantly higher in the model group than in the control group (all P < 0.001) , but there was no significant difference in IFN-γ expression ( P= 0.278) ; moreover, the model group showed significantly increased proportion of ILC2 (5.604% ± 2.105%) compared with the control group (1.750% ± 1.104%, P= 0.003) . In the Rag1 -/- mice, the ear skin was obviously red, swelling and dry with crusts in the model group, and HE staining showed increased epidermal thickness and eosinophil infiltration in the dermis; the model group showed significantly increased expression of IL-4, IL-5, thymic stromal lymphopoietin (TSLP) and IL-33 in skin tissues ( P= 0.010, 0.043, 0.034, 0.007, respectively) , but no significant difference in the expression of IL-13 or IFN-γ ( P= 0.274, 0.697, respectively) compared with the control group; the proportion of ILC2 was significantly higher in the model group (5.165% ± 2.436%) than in the control group (0.835% ± 0.578%, P= 0.014) ; the experimental group showed markedly attenuated skin lesions, reduced epidermal thickness and number of eosinophils infiltrating in the dermis, but no significant difference in the expression of IL-4, IL-5, IL-13, TSLP or IL-33 compared with the model group (all P > 0.05) . Conclusion:ILC2 play a role in the mice with AD-like inflammatory response induced by MC903, which dose not depend on adaptive immunity.

This paper is to report the study of the metabolism of forscolin in plasma and liver microsomes for guiding clinical therapy. Forscolin was quantified by HPLC-MS/MS. The metabolic stability of forscolin in rat, Beagle dog, monkey and human plasma and liver microsomes, mediated enzymes of forscolin and its inhibition on cytochrome P450 isoforms in human liver microsomes were studied. Results showed that forscolin was not metabolized in plasma of the four species but metabolized in liver microsomes of the four species. The t1/2 of forscolin in rat, Beagle dog, monkey and human liver microsomes were (52.0 +/- 15.0), (51.2 +/- 5.9), (6.0 +/- 0.2) and (11.9 +/- 1.8) min; CL(int) were (75.6 +/- 18.7), (60.9 +/- 6.8), (513.8 +/- 14.3) and (176.2 +/- 25.6) mL x min(-1) x kg(-1); CL were (34.8 +/- 4.5), (23.3 +/- 1.0), (40.3 +/- 0.5) and (17.9 +/- 0.3) mL x min(-1) x kg(-1), respectively. Forscolin was metabolized by CYP3A4 in human liver microsomes. There was definite inhibition on CYP3A4 at the concentrations of forscolin between 0.1 ng x mL(-1) and 5 microg x mL(-1). Therefore, forscolin is rapidly excreted from liver microsomes. Attention should be paid to the drug interaction when forscolin was used along with other drugs metabolized by CYP3A4 in clinics.
Animals ; Chromatography, High Pressure Liquid ; Coleus ; chemistry ; Colforsin ; blood ; isolation & purification ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Dogs ; Humans ; Macaca ; Metabolic Clearance Rate ; Microsomes, Liver ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Tandem Mass Spectrometry

The self—formulated modified“Blood—activating andQi—benefitting Decoction of Fructus Cirri Sarco-dactylis”with heavy dose of Radix Angelica Sinensiswas applied for the treatment of 50 cases of apoplexycomplicated with pseudo—Bulbar paralysis.The totaleffective rate of dysathria was 98% with a 58% abovemarked effect rate,while the total effective rate for-choking and coughing was 98%,with a 94% abovemarkedly effective rate.

In non-small cell lung cancer(NSCLC),as an improtant oncogenic driver gene,epidermal growth factor receptor exon 20 insertion(EGFR ex20ins)has a unique protein structure and is primarily drug-resistant to tradi-tional EGFR-tyrosine kinase inhibitors(EGFR-TKIs).In recent years,exploration of targeted therapy for EGFR ex20ins has never stopped.Firstly Mobocertinib and Amivantamab obtained approval from U.S.Food and Drug Administration(FDA)for EGFR ex20ins mutant NSCLC patients,then other drugs,such as Sunvozertinib,made breakthroughs and combination therapies also obtained gains.Multi-pronged measures are hopeful to overcome EGFR ex20ins drug resistance.As men-tioned above,it's definitely important to gain deeper understanding of molecular mechanism of EGFR ex20ins and assess ef-fect and difference between novel drugs.This review is devoted to make a summary about newest achievement so to provide valuable reference about precise therapy for patients with EGFR ex20ins.

AIMTo observe the effect of FAK-related non-Kinase (FRNK) plasmid on hepatic stellate cell (HSC) proliferation stimulated by fibronectin (FN).

METHODSFRNK plasmid was transfected into HSC with transient liposomal transfection. The proteins of FRNK, FAK and p-FAK(Tyr397) were assayed by Western blotting analysis. The proliferation of HSC was evaluated by improved MTT assay, and cell cycle pattern was determined by flow cytometry (FCM).

RESULTS(1) The expression of FRNK protein increased after FRNK transfected HSC, and it was at 48 h that the expression of FRNK protein was the highest (P < 0.01). The protein level of FAK was no significant difference between before FRNK plasmid transfection and after transfection (P > 0.05). The expression of p-FAK(Tyr397) protein was down-regulated after FRNK had been transfected in HSC, (P < 0.01). (2) The HSC proliferation inhibition rates at 12 h, 24 h and 48 h after FRNK transfection were 20.07%, 26.16%, 29.77%, respectively (P < 0.01). (3) Compared with the non-FRNK plasmid group, the FRNK-transfected HSCs almost arrested in G0/G1 phase (71.4 +/- 2.81 vs 48.9 +/- 1.66, P < 0.01).

CONCLUSIONAfter FRNK were transfected successfully in HSCs using lipofectamine, the phosphorylation of FAK was inhibited. The HSC proliferation was restrained in a time-dependent manner and the HSC was arrested in G0/G1 phase.

Cell Line ; Cell Proliferation ; Fibronectins ; Hepatic Stellate Cells ; cytology ; Humans ; Phosphorylation ; Plasmids ; genetics ; Protein-Tyrosine Kinases ; genetics ; Transfection