This study is to evaluate the HAART pharmacodynamics with dolutegravir as the "anchor" in vitro. A nucleoside reverse transcriptase inhibitors (NRTIs) resistant recombinant virus model (VSVG/HIV-1(RT-D67N,K70R,T215F)) and an integrase inhibitors (INIs) resistant recombinant virus model (VSVG/HIV-1(IN-G140S,QI48H)) were constructed and established. The anti-viral pharmacodynamics was evaluated with drug combinations including two NRTIs along with one INI or one NNRTI. The results showed that the combination with an INI gave a stronger synergism on wild type HIV-1 replication comparing to that with an NNRTI. Comparing the two INIs as the "anchor" for HAART, DTG exhibited an equivalent CI to that of RAL on wild type HIV-1 replication; but a greater synergy than RAL on INI-resistant HIV-1 replication. Besides of the pharmacodynamics results of DTG-based drug combination, the results may contribute to clinical antiviral therapy.
Antiretroviral Therapy, Highly Active ; Cells, Cultured ; Drug Resistance, Viral ; HIV Integrase Inhibitors ; pharmacology ; HIV-1 ; drug effects ; physiology ; Heterocyclic Compounds, 3-Ring ; pharmacology ; Humans ; Virus Replication ; drug effects

Objective To investigate the inhibitory effect of adriamycin in combination with hyperthermia on apoptosis and bcl-2 expression in the chronic leukemic cell line K562/AO2 in vitro.Methods The working con- centration of adriamycin against K562/AO2 determined by using methyl thiazolyl tetrazolium(MTT) assay was used to treat the chronic leukemic cell line K562/AO2 in vitro alone or in combination with hyperthermia induced using a hot water bath at 40,41 or 42℃.The inhibitory effect was evaluated by MTT assay.The apoptosis rates and bcl-2 ex- pression of K562/AO2 were determined by flow cytometry.Results The working concentration of adriamycin in the study was defined as its 50% inhibition concentration (IC50).A 60 min session of hyperthermia at 40℃,41℃or 42℃was associated with significant growth inhibition of the cell line K562/AO2.Adriamycin chemotherapy alone and with hyperthermia significantly inhibited the growth of K562/AO2.All treatments significantly increased apoptosis rates and down-regulated bcl-2 expression of the K562/AO2 cell line.Conclusion Adriamycin chemotherapy com- bined with 60 min sessions of hyperthermia showed significant suppression effect on K562/AO2 cell proliferation.The treatment can increase apoptosis rates and down-regulate bcl-2 expression.

Objective Kangfuling granules can be used for the treatment of radiation damage , but its quality criteria has not been established .This paper aimed to establish the quality criteria of Kangfuling granules to control its quality . Methods Radix as-tragali and Radix angelicae sinensis in the formula both were identified qualitatively by TLC .The content of astragaloside IV was exam-ined by HPLC:Agilent Zorbax Extend-C18 (4.6 mm ×250 mm, 5μm) was used as the chromatographic column .The mobile phase was consisted of acetonitrile-water (32:68) with a flow rate at 1.0 mL/min.The evaporative light scattering detector was used to detect the compound.The content of astragaloside IV and ferulaic acid was examined by HPLC: Agilent Zorbax SB-C18 (4.6 mm ×250 mm, 5 μm) was used as the chromatographic column .The mobile phase was consisted of acetonitrile-0.085% phosphoric acid water (17:83) with a flow rate at 1.0 mL/min.The wave length was 320 nm and column temperature was set at 35℃. Results The same color spot was shown in the chromatogram of the test sample and the control sample in the corresponding position with no interference of negative control.The linear range for Astragaloside IV 0.030 6 mg/mL~0.612 0 mg/mL with R2 =0.999.The RSD of stability test was 2.17%.The RSD of precision test was 1.89%.The RSD of re-peatability test was 1.58%.The average recovery was 101.26%. The linear range for ferulaic acid 0.24-4.80 μg/mL with R2 =0.999.The RSD of stability test was 1.37%.The RSD of precision test was 0.83%.The RSD of repeatability test was 1.14%.The average recovery was 98.39%.This product was tentatively calculat-ed according to the dry matter , the amount of Astragaloside IV and ferulaic acid should not be less than 0.19 mg/g and 0.08 mg/g, re-spectively. Conclusion In this study, the established TLC was used for the qualitative identification of Kangfuling granules , and the content of Astragaloside IV and ferulaic acid was not less than 0.19 mg and 0.08 mg in per gram of Kangfuling granules , respectively. The established standard is suitable for the quality control of Kangfuling granules .

AIM:To observe the effects of granulocyte colony-stimulating factor (G-CSF) on calcium, sodium and potassium ion channel currents of the ischemic atrial myocytes in guinea pig by whole-cell patch clamp technique.METHODS:The guinea pig atrial myocytes were obtained by enzymolysis.Under ischemia and hypoxia condition, whole-cell patch clamp was used to observe the effects of G-CSF at various concentrations on the changes of the I-V curve, activation curve and availability of L-type calcium channel current (ICa,L) and voltage-dependent sodium channel current (INa), as well as I-V curve of delayed rectifier potassium channel current (IK).RESULTS:Under ischemic condition, the I-V curves of ICa,L were changed by acute G-CSF intervention in a dose-dependent fashion.Except for G-CSF at dose of 300 μg/kg, the other concentrations of G-CSF did not change the activation curve and availability of ICa,L, indicating that the effects of G-CSF on ICa,L were in a voltage-independent fashion.The I-V curves of ICa,L under ischemic condition were gradually approaching the normal levels by the higher dose of G-CSF, while the effect of 300 μg/kg G-CSF on ICa,L was similar to 100 μg/kg G-CSF.Acute G-CSF intervention at different doses did not change I-V curve, activation curve, and availability or steady-state availability of INa.As a part of IK, the rapid activating component (IKr) was improved by 100 μg/kg and 300 μg/kg G-CSF intervention with the similar effects, while the slowly activating component (IKs) was not changed by G-CSF.CONCLUSION:G-CSF affects ion channel electrophysiological properties of ischemic atrial myocytes in a voltage-independent but concentration-dependent manner, thus reducing the incidence of atrial arrhythmia.

The traditional Chinese medicine (Tripterygium wilfordiiHook.f.,TWH) has been clinically used to treat primary and secondary renal diseases and proteinuria for nearly 40 years.However,there is a rare literature about the effect of triptolide (the main active ingredient of TWH) on the expression of oxidative carbonyl protein (OCP) in diabetic nephropathy (DN).This study aimed to provide experimental evidence for triptolide treatment on DN through its effect on the expression of OCP,in order to investigate the effects of triptolide on the expression of OCP in rats with DN.Sixty SD rats were randomly divided into five groups:control group,high-dose triptolide (Th) group,low-dose triptolide (T1) group,DN model group,and positive control (benazepril) group.The DN model was established using streptozotocin.Urinary protein excretion,fasting blood glucose (FBG),superoxide dismutase (SOD) in renal homogenate,malondialdehyde (MDA) in renal homogenate and renal nitrotyrosine by immunohistochemistry,and the expression of OCP by oxyblotimmune blotting were detected.In the DN model group,rat urinary protein excretion and renal MDA were significantly increased,while renal SOD significantly decreased and nitrotyrosine expression was obviously upregulated in the kidney.After triptolide treatment,24-h urinary protein excretion (61.96±19.00 vs.18.32±4.78 mg/day,P＜0.001),renal MDA (8.09±0.79 vs.5.45±0.68 nmol/L,P＜0.001),and nitrotyrosine expression were decreased.Furthermore,renal OCP significantly decreased,while renal SOD (82.50±19.10 vs.124.00±20.52 U/L,P＜0.001) was elevated.This study revealed that triptolide can down-regulate the expression of OCP in the renal cortex of DN rats.

Acute pancreatitis (AP) is an inflammatory disease of the pancreas. Its pathogenesis is not only related to abnormal activation of trypsinogen, but also related to calcium overload, mitochondrial dysfunction, impaired autophagy and endoplasmic reticulum stress. However, the mechanism has not been fully elucidated and needs to be further studied. Currently, there is no effective treatment for AP. It is difficult to prevent the loss of pancreatic function. An in-depth understanding of the pathophysiological mechanisms of AP may help to identify the potential therapeutic targets. Therefore, the purpose of this study is to review recent advances in the mechanism of AP in order to provide more research direction for treatment.

Objective To investigate the diagnosis value of the tissue strain imaging in myocardial dysfunction in systemic lupus erythematosus (SLE). Methods Sixty-two patients and sixty controls underwent conventional and tissue Doppler echocardiography. Peak strain and strain rate value during systolic and diastolic phases as well as E/A, left ventricular fraction shortening (LVFS), left ventricular ejection fraction(LVEF) were measured in both SLE and the control groups. Results ①E/A,LVFS and LVEF did not differ between SLE patients and controls( P ＞0.05). The systolic peak strain and strain rate of SLE patients were lower than those of controls but without significant differences( P ＞0.05). ②The diastolic peak strain and strain rate of SLE patients were significantly lower than those of controls (P ＜0.01). ③ The diastolic peak strain and strain rate of antiheart antibody (AHA) positive patients were significantly lower than those of negative ones( P ＜0.05). Conclusions Strain and strain rate combining with AHA can sensitively detect myocardial dysfunction of SLE.

In order to study the spectral reflectance differences of Glycyrrhizae Radix under different growth conditions and lay the foundation for quantitative monitoring of Glycyrrhizae Radix remote sensing images, spectra of Glycyrrhiza species under different growth period and different varieties and different regions were measured by a portable spectrometer. The results showed that the reflectivity of annual G. uralensis was obviously higher than that of the two years plant in the visible light band own to the contents of crown layer chlorophyll. The reflectivity of two years G. pallidiflora was higher than that of G. uralensis in the near infrared band own to the leaf area index and the content of leaf water. The red edge spectrum of annual plant fluctuated largely than that of two years plant due to vegetation coverage and leaf area index. G. pallidiflora grew well than G. uralensis. Under different regions of the Glycyrrhiza species, spectral data analysis showed that within a certain range, the average annual precipitation and average annual evaporation were the major factors to affect the differences of Glycyrrhiza species spectral data under different regions owe to the leaf water content, the higher leaf water content, the lower spectral reflectance. The principal component analysis and continuum-removed method of the spectral data under different regions found that, within a certain range, the average annual precipitation and average annual evaporation were the major factors caused by the differences of Glycyrrhiza species spectral data under the different regions, Glycyrrhiza species spectral similarity related to the spatial distance.
Geography ; Glycyrrhiza ; chemistry ; Principal Component Analysis ; Spectrum Analysis

OBJECTIVETo analyze the changes of inhibitory killer cell immunoglobulin-like receptors (KIRs), NKG2D receptor and the cytotoxicity of natural killer (NK) cells induced by persistent exposure to CNE2 cells.

METHODSThe HLA-class I genotypes of CNE2 cells and KIR genotypes were determined by PCR with sequence-specific primers (PCR-SSP). The expressions of KIR2DL1, KIR2DL3, KIR3DL1, and NKG2D by the NK cells (freshly isolated NK cells, NK cells cocultured with 100 U/ml IL2 or with 100 U/ml IL2 and CNE2 cells as the control, IL2 and CNE2 groups, respectively) were analyzed by flow cytometry. Cytotoxicity of NK cells against CNE2 cells were detected by LDH releasing assay.

RESULTSThe HLA genotypes of CNE2 cells were A2, 24, B18, 35, Cw4, 7. NK cells isolated from 3 healthy donors expressed KIR2DL1, KIR2DL3, and KIR3DL1. After 4, 24 and 48 h of culture, NK cells in CNE2 group displayed higher KIR2DL1, KIR2DL3 but lower NKG2D expression than those in the control and IL2 groups (P<0.01), whereas the latter two groups showed no significant difference in KIR2DL1, KIR2DL3, and NKG2D expressions (P>0.05), and no difference in KIR3DL1 expression was found between the 3 groups (P>0.05). After 24 h of culture, the cytotoxicity against CNE2 cells mediated by the NK cells in IL2 and CNE2 groups were (26.96-/+1.47) % and (2.74-/+1.64) % at E:T ratios of 10:1, and (35.74-/+3.59)% and (4.57-/+2.41) % at E:T ratio of 20:1, respectively. NK cells in CNE2 group displayed lower cytotoxicity than those in IL2 group (P<0.01).

CONCLUSIONSPersistent exposure to tumor cells expressing NKG2D ligands can lead to downregulated expression of NKG2D receptor, increased expression of KIRs and reduction of NK-mediated cytolysis. These results elucidate the molecular mechanism of reduced cytotoxicity mediated by the edited NK cells and indicate that blocking HLA-class I-bound KIRs or enhancing the expression of NKG2D may promote NK cell-mediated cytolysis.

Cell Line, Tumor ; Cell Survival ; immunology ; Cytotoxicity, Immunologic ; immunology ; Flow Cytometry ; Humans ; Killer Cells, Natural ; cytology ; immunology ; metabolism ; NK Cell Lectin-Like Receptor Subfamily K ; Nasopharyngeal Neoplasms ; immunology ; metabolism ; pathology ; Receptors, Immunologic ; metabolism ; Receptors, KIR ; metabolism ; Receptors, KIR2DL1 ; metabolism ; Receptors, KIR2DL3 ; metabolism ; Receptors, Natural Killer Cell

OBJECTIVETo compare clinical outcomes between two suturing methods using non absorbable materials through drilling the bone and suturing anchors for the treatment of complete rupture of the deltoid ligament.

METHODSFrom January 2009 to January 2013, 58 hospitalized patients with ankle fracture combined with complete rupture of the deltoid ligament were treated with suturing using non absorbable materials through drilling the bone or suturing anchors. There were 29 patients who received suturing treatments using non absorbable materials through drilling the bone (Group A), including 18 males and 11 females, with an average age of (39.76 +/- 11.81) years old. According to the Lauge-Hansen classification, 12 patients had supination external rotation (SER) injuries with IV degree, 5 patients had pronation external rotation (PER) injuries with III degree, 10 patients had PER injuries with IV degrss, and 2 patients had pronation abduction injuries with III degree. There were 29 patients who received treatments with suturing using anchors (Group B), including 14 males and 15 females, with an average age of (41.79 +/- 13.28) years old. According to the Lauge-Hansen classification,9 patients had SER injuries with IV degree, 6 patients had PER injuries with III degree,13 patients had PER injuries with IV degree, and 1 patient had pronation abduction injuries with III degree. All the patients were treated with open reduction and internal fixation, as well as reconstruction of deltoid ligaments to restore the stability of the medial ankle structures. The clinical examination, imaging evaluation, American society for ankle surgery (AOFAS) ankle-hindfoot score and visual analogue scale (VAS) were used to evaluate the clinical results after operation, and the results of the two groups were compared and analyzed statistically.

RESULTSThe follow-up duration of the 58 patients ranged from 23 to 40 months,with an average of 27.3 months. All the patients had fracture union, and the mean healing time was 12.3 weeks (ranged, 10 to 17 weeks). There were no incision complications and ankle instability. There were no significant differences between two groups in AOFAS (P=0.666) and the VAS (P=0.905).

CONCLUSIONTreatments of complete rupture of the deltiod ligaments with the two suturing methods get similar good clinical effects, but the suturing using non absorbable materials through drilling the bone has several advantages such as reducing the financial burden of patients, saving social medical resources and avoiding the shortcoming in difficult removal of anchor suture.

Adolescent ; Adult ; Ankle Fractures ; surgery ; Ankle Injuries ; surgery ; Ankle Joint ; surgery ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; Humans ; Lateral Ligament, Ankle ; injuries ; surgery ; Male ; Middle Aged ; Young Adult