Erlotinib,a kind of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs),has been effectively used in the treatment of non-small cell lung cancer (NSCLC).Although it prolongs patients survival time,erlotinib is limited to be further applied for its resistance.It has been proved that threonine to methionine mutations in codon 790 (T790M),Kirsten rat sarcoma viral oncogene homolog (KRAS) mutation and the amplification of met oncogene play important roles in the drug resistance.Based on the different molecular mechanisms of resistance,multiple clinical trials of the second generation TKIs,retreatment of chemotherapy or erlotinib and subsequent treatment according to failure modes have been developed.

Breast Neoplasms ; diagnosis ; Diffusion Magnetic Resonance Imaging ; Female ; Humans

Objectives To investigate the effect of proliferation and invasiveness by turmeri cvolatile oil on human neuroblastoma cell line SH-SY5Y. Methods Cells were incubated with different concentrations of TVO in vitro. Then cell survival rate was measured by MTT assay. The effect of 160 mg/L TVO on cell migration was assessed by cell scuffing test. Invasive ability of cell was detected by Transwell test. Apoptosis of cells was detected observed by flow cytometry assay. Results Survival rate of SH-SY5Y cells decreased and apoptisis rate was abated with elevated TVO concentration and prolonged cultivation time. Level of cell migration was lower than that in control group after being cultured with 160 mg/L TVO solution for 12 , 24 and 48h. With the in-crease of TVO concentration , the invasion ability of cells gradually decreased , and the invasive force and cis-platin had no obvious difference when the concentration of drug reached 160 mg/L. Conclusion The prolifera-tion of cells can be inhibited by inhibiting the proliferation and invasiveness ability with TVO.

Objective To evaluate the effects of turmeric volatile oil (TVO) combined with cisplatin on the proliferation and apoptosis of a human cutaneous squamous cell carcinoma cell line A431,and to explore their mechanisms.Methods Some cultured A431 cells at exponential growth phase were divided into several groups to be treated with 5,10,20,40 and 80 mg/L TVO,as well as high-glucose Dulbecco's modified Eagle's medium (DMEM) containing 1％ dimethyl sulfoxide (DMSO,control group),respectively.After 24-hour treatment,cell counting kit 8 (CCK8) assay was performed to estimate the proliferative activity of A431 cells in the above groups.Some other A431 cells were divided into 4 groups:control group treated with high-glucose DMEM containing 1％ DMSO,TVO group treated with 40 mg/LTVO,cisplatin group treated with 10 mg/L cisplatin,and TVO + cisplatin group treated with 40 mg/L TVO and 10 mg/L cisplatin.After 24-hour treatment,CCK8 assay was performed to estimate the cellular proliferative activity,inverted microscopy to observe changes in cell morphology,fluorescence microscopy to detect cell apoptosis after acridine orange (AO)/ethidium bromide (EB) double-staining,colorimetry to evaluate the activity of Caspase-3 and Caspase-9,and Western blot analysis to determine the protein expression of Caspase-3 and p-glycoprotein.Results After 24-hour treatment with 5,10,20,40 and 80 mg/L TVO,the cell proliferation rates were inhibited by (12.83 ± 6.4)％,(16.27 ± 11.4)％,(21.61 ± 9.1)％,(33.11 ± 2.0)％ and (46.00 ± 3.3)％ respectively,and the inhibition rates were all significantly higher in these groups than in the control group (4.03％ ± 1.4％,all P ＜ 0.05).The 50％ inhibitory concentration (IC50) of TVO at 24 hours was (61.66 ± 1.03) mg/L.Compared with the control group,the proliferation inhibition rates significantly increased in the TVO group,cisplatin group and TVO + cisplatin group (all P ＜ 0.05),suggesting that the combination of TVO and cisplatin showed synergistic inhibitory effects with a combination index of 1.366.Moreover,A431 cells turned round to different extents and became apoptotic in the TVO group and cisplatin group,and the TVO + cisplatin group showed obviously decreased number of cells and a large number of cell debris.The TVO + cisplatin group also showed significantly increased activity of Caspase-3 (1.520 ± 0.115) and Caspase-9 (2.760 ± 0.297) as well as protein expression of Caspase-3 (1.482 ± 0.016) compared with the TVO group (Caspase-3 activity:1.117 ± 0.095;Caspase-9 activity:1.259 ± 0.059;Caspase-3 protein expression:1.156 ± 0.006,all P ＜ 0.01) and cisplatin group (Caspase-3 activity:1.381 ± 0.089;Caspase-9 activity:1.829 ± 0.171;Caspase-3 protein expression:1.296 ± 0.021,all P ＜ 0.01),but significantly decreased p-glycoprotein expression (0.528 ± 0.014) compared with the TVO group (1.311 ± 0.011,P ＜ 0.01) and cisplatin group (1.169 ± 0.012,P ＜ 0.01).Conclusion TVO combined with cisplatin can synergistically inhibit the proliferation of A431 cells and induce cell apoptosis,which may be associated with activation of the caspase system and decreased expression of pglycoprotein.

Objective To study the therapeutic effects of Ag85A plasmid DNA vaccines in a mouse model of multi-drug resistant-(MDR-) Mycobacterium tuberculosis infection. Methods BALB/c mice were infected with Mycobacterium tuberculosis clinical strain HB361 with isoniazid and rifampin resist-ance by intratail-vein injection and were subsequently divided into 6 groups. At the third day after infection, the mice were treated with saline (group A), vector pVAX1 (greup B), rifampin (group C), vaccae (group D), Ag85A plasmid DNA vaccines (group E),rifampin and Ag85A plasmid DNA vaccines (group F) for 60 d. The lungs and spleens from the mice were taken and their pathological changes, weight and number of myeobacterial colony were examined at the third week after the end of treatment. Results At third week af-ter the end of treatment, the gross pathological observation and histopathological examination in lung showed that the lung lesions were limited, the profile of the alveoli was relatively clear, and normal structure could be seen in 2/3 areas of the lung sections in group D, E and F. The extent of lung lesion was 50% in group D,20% in group E and F. The pathological changes in group A, B, and C were more severer than those in group D, E and F. Compared with group A, the colony-forming units (CFU) in the lungs from mice in group D,E and F decreased 52%, 68%, 78%, respectively. The CFU in the spleens from mice in group D,E and F decreased 48%, 65%, 79%, respectively. Conclusion Ag85A plasmid DNA vaccines alone or Ag85A plasmid DNA vaccines along with chemotherapy have significant therapeutic effects on the mouse model of MDR-Mycobacterium tuberculosis infection.

Objective To explore the clinical effects of transvaginal acupoint injection in the treatment of chronic pelvic pain induced by pelvic floor myofascitis. Methods All 80 patients with pelvic floor myofascitis were randomly assigned to an observation group and a control group, with 40 patients in each group. The observation group was given the treatment of transvaginal pain spot injection, and the control group was given transvaginal non-pain spot injection. Changes of clinical symptoms, signs, pain scores and other indices before the treatment, 1 week and 2 weeks after the treatment between the two groups of patients were observed and compared. Results Comparison of pain scores between the treatment group and the control group before the treatment was not significantly different (P>0.05); pain scores after the treatment in the two groups were significantly lower compared to those before the treatment, and the differences in the two groups were both significant (P<0.01). The curative effects in the treatment group were better than those in the control group (P<0.01). Conclusion Transvaginal acupoint injection is able to significantly alleviate chronic pelvic pain induced by pelvic floor myofascitis, but accurate pain spot injection is required.

OBJECTIVETo study the correlation between the MRI apparent diffusion coefficient (ADC) value and histological grade and molecular biology of breast invasive ductal carcinoma (IDC).

METHODSThis retrospective study included 125 patients with IDC verified by pathology from February 2010 to February 2013. Conventional MRI and diffusion-weighted imaging (DWI) examination were performed using a 3.0T scanner with diffusion factor of 0 and 800 s/mm(2). The region of interest (ROI) was drawn on the largest lesion and/or its two adjacent slices. The ADC value of the whole tumor was calculated as the mean ADC value. The correlation between mean ADCs and histological grade and biological factors was analyzed.

RESULTSThe mean ADC of pathological grade I, II and III IDC was (1.152 ± 0.072)×10(-3) mm(2)/s, (1.102 ± 0.101)×10(-3) mm(2)/s, and (1.035 ± 0.107)×10(-3) mm(2)/s, respectively. There was a statistically significant difference among them (P = 0.003). Statistically a significant difference was observed between grade III and I (P = 0.034), grade III and II (P = 0.006), but not between grade I and II (P = 0.741). A significant correlation was observed between ADC value and pathological grade (r = -0.342, P < 0.001). The median ADC values were significantly higher in the ER-negative than in the ER-positive cases [(1.130 ± 0.115)×10(-3) mm(2)/s vs. (1.060 ± 0.089) ×10(-3) mm(2)/s, P < 0.001)], in PR-negative than in PR-positive cases [(1.121 ± 0.106)×10(-3) mm(2)/s vs. (1.055 ± 0.096) ×10(-3) mm(2)/s, P < 0.001)], and in Ki-67-negative than in Ki-67-positive cases [(1.153 ± 0.090)×10(-3) mm(2)/s vs. (1.063 ± 0.101) ×10(-3) mm(2)/s, P < 0.001]. A statistically significant correlation was observed between ADC value and expressions of ER, PR, and Ki-67 (r = -0.311, r = -0.317, r = -0.414, P < 0.001).

CONCLUSIONADC value of breast invasive ductal carcinoma is correlated with histological grade, and expression of ER, PR and Ki-67.

Objective:To study the intervention effect of ganoderma triterpenoids combined with exogenous monosialotetrahexosyl ganglioside(GM1) on cognitive dysfunction and synaptic ultrastructure of hippocampal neurons in rats with epilepsy caused by pentylenetetrazol(PTZ).Methods:A total of 40 Sprague-Dawley rats were divided into blank control group, epileptic model group, ganoderma triterpenoids group, GM1 group and GM1 combined with ganoderma triterpenoids group according to the random number table method( n=8 in each group). The rats were intraperitoneally injected with PTZ subconvulsant dose (35 mg·kg -1·d -1) once a day for 28 days to replicate the models of chronic epilepsy. And the rats in different medication groups were given corresponding administration based on daily intraperitoneal injection of PTZ(GM1: intraperitoneal injection of 30 mg·kg -1·d -1, ganoderma triterpenoids: gavage 1 000 mg·kg -1·d -1). Morris water maze was used to test the spatial exploration and learning and memory ability of epileptic rats.Transmission electron microscopy was used to observe the ultrastructure of hippocampal neurons in epileptic rats.Immunofluorescence staining was used to observe expression levels of cofilin and SYN protein in hippocampus CA1 of rats. In addition, Western blot was used to detect the expression levels of cofilin, p-cofilin and synaptophysin(SYN) protein in hippocampus of rats. SPSS 17.0 software was used for statistical analysis. Repeated one-way ANOVA was used for comparing among groups, LSD test was used for pairwise comparisons. Results:Morris water maze results showed that there were statistically significant differences in escape latency, times of crossing the platform and time spent in the target quadrant among the groups( F=5.259, 8.240, 5.961, all P<0.05). Compared with the epilepsy model group, the escape latencies((20.31±7.39) s, (21.81±6.05) s, (17.66±4.76) s) of the ganoderma triterpenoids group, GM1 group and GM1 combined with ganoderma triterpenoids group were shorter (all P<0.05), the numbers of crossing the platform ((4.63±1.41) times, (4.50±1.93) times, (5.50±1.77) times) were more (all P<0.05), the residence time in target quadrant ((31.91±5.00) s, (30.49±5.72) s, (35.70±5.34) s) were longer (all P<0.05). And the most obvious change was found in the GM1 combined with ganoderma triterpenoids group ( P<0.01). The results of transmission electron microscope showed that there were significant differences in the numbers of hippocampal neurons synapses, the synaptic gap, the density of postsynaptic membrane and length of active area of postsynaptic membrane among the groups( F=3.693, 7.201, 5.012, 4.033, all P<0.05). Compared with the epilepsy model group, the numbers of synapses ((8.00±1.79), (7.83±1.84), (8.50±1.87)) in the ganoderma triterpenoids group, GM1 group and GM1 combined with ganoderma triterpenoids group were all more (all P<0.05), synaptic gap ((33.83±3.81)nm, (32.43±4.14)nm, (30.23±3.08)nm)were narrower, and the postsynaptic dense substances ((57.50±6.03)nm, (58.10±2.40)nm, (60.73±3.81)nm) were all thicker (all P<0.05). The length of active region of postsynaptic membrane ((271.66±11.80) nm, (279.06±13.58) nm) in ganoderma triterpenoid group and GM1 combined with ganoderma triterpenoids group were longer than that in epilepsy model group (both P<0.05). Immunofluorescence results showed that the average fluorescence intensity of cofilin in the epilepsy model group was higher than that in the blank control group, and the average fluorescence intensity of SYN was lower than that in the blank control group (both P<0.05). The average fluorescence intensity of cofilin in GM1 group and GM1 combined with ganoderma triterpenoids group were lower than that in epilepsy model group (both P<0.05), and the average fluorescence intensity of SYN in ganoderma lucidum triterpenoids combined with GM1 group was higher than that in epilepsy model group ( P<0.05). Western blot showed that the expression levels of cofilin protein in the epilepsy model group was higher than that in the blank control group ((1.454±0.080), (1.092±0.099), P<0.05), and the expression of p-cofilin and SYN were lower than those in the blank control group ((1.103±0.120) vs (1.420±0.934), (1.650±0.062) vs (1.958±0.062), both P<0.05). The expression of cofilin protein ((1.227±0.071), (1.262±0.078), (1.162±0.129), P<0.05) in ganoderma triterpenoids group, GM1 group and GM1 combined with ganoderma triterpenoids group were lower than that in epilepsy model group, and the expression levels of p-cofilin(1.357±0.199) and SYN protein(1.873±0.010) in ganoderma triterpenoids combined with GM1 group were higher than that in epilepsy model group (both P<0.05). Compared with ganoderma lucidum triterpenoids group and GM1 group, there was no significant difference in each index of GM1 combined with ganoderma triterpenoids group (all P>0.05). Conclusion:GM1 combined with ganoderma triterpenoids may promote the synaptic plasticity of neurons, improve the learning and memory ability of epileptic rats.Combination medication is better than single medication in some observed indicators.

Objective:To investigate the serum levels of soluble growth stimulation expression gene 2 protein (sST2) and inflammatory factors in patients with acute left ventricular ejection fraction reduction heart failure (HFrEF) treated with sacubitril/valsartan.Methods:Ninety six patients with acute HFrEF admitted in The Affiliated Hospital of Qingdao University from March 2020 to March 2021 were enrolled. The patients were treated with sacubitril/valsartan,the dose was gradually increased from 50 mg b.i.d to the target dose of 200 mg b.i.d according to hemodynamics. After 12 weeks, the target dose was achieved in 72 cases (compliance group), and did not achieved in 24 cases (non-compliance group). The serum levels of sST2, IL-1β, IL-6, TNF-αand IL-10 were measured and compared between the two groups. The changes in left atrial anteroposterial diameter (LA), left ventricular end-diastolic diameter (LVDd) and left ventricular ejection fraction (LVEF) values were assessed with echocardiography. The adverse reactions, readmission rate and all-cause death within 3 months after discharge were compared between the two groups.Results:A total of 96 patients with acute HFrEF completed the follow-up, including 72 patients (75.0%) in the compliance group and 24 (25.0%) in the non-compliance group; aged 50-75 (66.1±6.7) years old, and 68 (70.8%) males. After treatment, the serum levels of sST2, IL-1β, IL-6 and TNF-α were decreased, and the IL-10 level was increased in both groups ( P<0.05); while the improvement of serum indicators in the compliance group was more marked ( P<0.05). Echocardiography showed that the LA, LVDd, and LVEF were significantly increased after treatment ( P<0.05) in compliance group, while there was no significant changes before and after treatment in the non-compliance group. SST2, inflammatory factors and echocardiographic measurements of patients in the standard group had statistical significance before and after treatment ( P<0.05), and the difference showed a downward trend. No deterioration of renal function and angioedema were observed in both groups, and there was no significant difference in hyperkalemia (two in compliance group and one in non-compliance group), symptom hypotension (each in two groups) between the two groups (χ 2=0.12, 0.68; P>0.05). In the non-compliance group, 10 patients (41.7%) were readmitted due to heart failure, and 6 patients (25.0%) died; while there were no readmitted cases or fatal cases in compliance group (χ 2=33.49, 19.20; P<0.05). Conclusion:Early application of sacubitril and valsartan sodium in patients with acute HFrEF after hemodynamic stabilization can significantly improve left ventricular remodeling, for those with earlier escalation to the target dose, it is more beneficial. The changes of serum sST2 and inflammatory factor level after treatment may predict the efficacy of sacubitril/valsartan therapy.