Advances in Diagnosis and Treatment of HER2-positive Non-small Cell Lung Cancer / 中国肺癌杂志

Lung cancer is the most common malignancy in the world and the leading cause of cancer death. Human epidermal growth factor receptor 2 (HER2) positive non-small cell lung cancer (NSCLC) refers to the NSCLC caused by mutation, amplification or overexpression of the HER2 gene, resulting in its dysfunction. HER2 is the most active receptor in the HER family and can combine with other members to form dimers, which can activate multiple signaling pathways and regulate cell proliferation, differentiation, migration and apoptosis. In NSCLC, HER2 positivity is usually considered a poor prognostic marker. At present, the diagnosis and treatment of HER2-positive NSCLC are not mature. Immunohistochemistry (IHC), next generation sequencing (NGS) and other technologies are often used to detect the positive status of HER2 mutation, amplification or overexpression. In previous studies, antitumor drugs did not show ideal therapeutic effects in HER2-positive NSCLC. However, in recent years, related researches have shown that antibody-drug conjugates (ADCs) and new tyrosine kinase inhibitors (TKIs) in targeted therapy show good antitumor activity against HER2 positive NSCLC. This article summarized the progress in diagnosis and treatment of HER2-positive NSCLC, so as to provide reference for subsequent researches.
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Clinicopathological Characteristics and Therapeutic Effect of Patients with Non-small Cell Lung Cancer and Uncommon EGFR Mutations / 肿瘤防治研究

Objective To investigate the clinicopathological characteristics and treatment effect of patients with non-small cell lung cancer (NSCLC) and uncommon epidermal growth factor receptor (EGFR) gene mutations. Methods Real-time fluorescence quantitative PCR was used to detect the mutation of EGFR in 674 samples of patients with NSCLC.The correlation between uncommon EGFR mutations and clinicopathological characteristics was analyzed. Results The EGFR mutation rate was 47.92%, of which the incidence of uncommon EGFR mutations was 5.19%, showed the presence of ex18 G719 A/S/C (G719X)(1.63%), ex20ins (1.04%), ex21 L861Q (0.74%), and compound mutations (1.78%).Correlation analysis showed that uncommon EGFR mutations were more common in women, non-smokers, patients with high-medium differentiation and adenocarcinoma, and patients were more prone to brain and bone metastasis (all P < 0.05).NSCLC with uncommon EGFR mutations showed no significant differences in clinical and pathological features compared with those with common sensitive mutations (all P > 0.05).Follow-up information was available on 31 patients, with a median follow-up time of 10 months, of which 23 were in advanced stage.Among eight patients with G719X mutation in late stage, seven patients used EGFR tyrosine kinase inhibitor (EGFR TKIs)(five of them used afatinib) in the first line and had a median PFS of 12 months; one patient received chemotherapy with pemetrexed and carboplatin and had PFS of seven months, which was lower than that of the TKI group.Among four patients with L861Q mutation in late stage, one patient was untreated and the three remaining were treated with TKI in the first line and had a median PFS of eight months.The patient who was treated with afatinib and bevacizumab was still stable after 11 months of follow-up.Two patients with EGFR ex20ins in advanced stage were treated with chemotherapy and bevacizumab.Nine patients with compound mutations in advanced stage were treated with TKI; among which, five patients harboring T790M compound mutations were treated with third-generation TKI and had a median PFS of more than 10 months. Conclusion The correlation between specific uncommon EGFR mutation and clinical pathological characteristics varies.For advanced patients with uncommon EGFR mutations (except for ex20ins), TKI is generally chosen as the first-line clinical treatment.Afatinib is recommended for advanced NSCLC patients with G719X and L861Q mutations.Third-generation TKI has significant efficacy in patients with complex mutations containing T790M.

Overview of Multiplex Immunohistochemistry and Immunofluorescence Techniques in the Lung Cancer Immunotherapy / 中国肺癌杂志

Lung cancer is the most common malignancy and is a major public health problem worldwide. Programmed cell death receptor 1 (PD-1)/programmed cell death ligand 1 (PD-L1) inhibitors provide a new treatment strategy for non-small cell lung cancer (NSCLC). The existing biomarkers all have advantages and defects in selecting patients who benefit from immunotherapy. The multiplex immunohistochemistry/immunofluorescence (mIHC/IF) provides multiplex staining, allows comprehensive studies of cellular composition, cellular functions and cell-cell interactions. A number of studies have used mIHC/IF to explore specific immune cells in tumor immune microenvironment (TIME) and found that it is helpful for clinical prognosis and efficacy prediction in patients with lung cancer. In the era of immunotherapy for lung cancer, this technique has a bright future in translational research and clinical practice. The research progress of mIHC/IF detection in lung cancer immunotherapy is summarized in this review.
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Clinical significance of multi-sample next-generation sequencing for detection of gene mutations in non-small cell lung cancer / 肿瘤研究与临床

Objective:To explore the clinical significance of multi-sample next-generation sequencing (NGS) in detecting gene mutations in non-small cell lung cancer (NSCLC), so as to provide a basis for individualized targeted treatment.Methods:The data of 51 patients with NSCLC in the First Affiliated Hospital of Zhejiang University Medical College from June 2016 to February 2019 was retrospectively analyzed. The patients' age, gender, smoking status, pathological type, tumor staging, pleural invasion status and meningeal invasion status were collected, and the tissues, cerebrospinal fluid, pleural effusion and plasma samples were detected by NGS. The correlations between the driver gene mutations [epidermal growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK), and ROS proto-oncogene 1, receptor tyrosine kinase (ROS1)] and the clinicopathological features of patients were analyzed by χ 2 test or Fisher exact probability method. Cohen Kappa coefficient analysis was used to compare the consistency of NGS detection results in different samples. Results:Among the 51 patients, 3 different types of specimens were examined in 7 patients. Of which, the driver gene mutations were all positive in 3 cerebrospinal fluid samples and double gene mutations were found in 2 hydrothorax samples. Driver gene mutations were more common in female [90.48% (19/21) vs. 50.00% (15/30), χ 2 = 9.107, P = 0.003], non-smokers [80.00% (24/30) vs. 47.62 (10/21), χ 2 = 5.829, P = 0.016], adenocarcinoma patients [72.34% (34/47) vs. 0, P = 0.017] and patients with malignant pleural effusion [92.86% (13/14) vs. 56.76% (21/37), χ 2 = 4.443, P = 0.035], and the differences were statistically significant. However, there was no statistically significant difference in mutations of driver gene among patients with different age or tumor stages (both P > 0.05). The consistency rate of genetic test results of the driver gene mutations in 37 plasma samples and matched tissue samples was 70.27% (26/37), κ value was 0.430, which suggested a good consistency between them. Conclusions:In patients with advanced NSCLC, gene detection of cerebrospinal fluid is of high application value for patients with meningeal metastasis; for patients with pleural invasion, gene detection of pleural effusion is an effective means to screen targeted therapy drugs. NGS detection of multiple specimens can better reflect the status of gene mutations and guide the individualized targeted therapy of lung cancer patients.

Mechanism of promoted neovascularization by SVF after fat grafting / 中华整形外科杂志

Stromal vascular fraction(SVF)are the remaining cells after removing mature fat cells in the adipose tissue. Containing a certain amount of adipose derived stem cells(ADSCs), SVF also includes many other cells, which may have the potential of promoting angiogenesis. In this review, the role of SVF in angiogenesis after fat transplantation was summarized by intensive reading relative literature in recent years. The result is that angiogenesis and fat graft revascularization are regulated by various factors: SVF promotes secretion of a diverse array of cytokines and growth which are capable of stabilizing endothelium vascular network. ADSCs have the potential of differentiating into smooth muscle cells and endothelial cells which can coroperate to form new blood vessels.

Expression of leukocyte immunoglobulin-like receptor A3 in CD14 monocytes of patients with rheumatoid arthritis / 浙江大学学报·医学版

OBJECTIVE@#To investigate the expression of leukocyte immunoglobulin-like receptor A3 (LILRA3) in CD14 monocytes of patients with rheumatoid arthritis (RA).@*METHODS@#Fifty three RA patients admitted in the Second Affiliated Hospital of Zhejiang University School of Medicine from February 2017 to August 2017, and 21 healthy subjects were enrolled in the study. The expression of LILRA3 in CD14 monocyte subset was determined by flow cytometry, and its correlations with clinical features, laboratory examination results, antibodies and disease activity were analyzed.@*RESULTS@#LILRA3 percentage in the CD14 monocyte subset of RA patients was higher than that in the healthy controls (0.05). In addition, the percentages of LILRA3 in the monocytes of rheumatoid factor (RF)-positive or anti-cyclic citrullinated peptide (CCP) antibody-positive patients were significantly higher than those of the RF-or anti-CCP antibody-negative patients (all 5.1 was higher than that in patients with DAS28 ≤ 5.1 (<0.05).@*CONCLUSIONS@#The expression of LILRA3 is up-regulated in CD14 monocyte subset isolated from RA patients, and it is correlated with disease activity.

Effect of low-dose insulin on spinal N-methyl-D-aspartate receptor levels and the downstream signals in streptozotocin-induced diabetic rats / 中华老年医学杂志

Objective To investigate the effect and its significance of low-dose insulin on Nmethyl-D-aspartate receptor (NMDAR) level and its downstream signaling of c-Jun N-terminal kinase (JNK) and the extracellular signal-regulated protein kinase (ERK,p44/42MAPK) in streptozotocininduced diabetic rats.Methods Ten-week-old male SD rats were randomly divided into 3 groups:control group,diabetic group and insulin-treated diabetes group.Diabetes was induced by streptozocin (STZ,60mg/kg) injected intraperitoneally.Two weeks after STZ injection,insulin glargine (92u/day for 8 weeks) was subcutaneously injected in the insulin-treated diabetes group.Then,the rat lumbar spinal cords were collected.The protein levels of phospho-Insulin receptor substrate 1 (P-IRS1),phospho-NMDAR NR1 subunit (P-NR1),P-JNK and P-p44/42MAPK were evaluated by Western blotting.One week before the terminal of the study,paw thermal response latency was measured in all groups.Results Blood glucose levels were tremendously high in both the diabetic group and insulintreated diabetes group.Compared with the control group,paw thermal response latency was markedly shortened in the diabetic group (P＜ 0.001) and the insulin-treated diabetes group (P＜ 0.001),and the alteration was more pronounced in the diabetic group (P＜0.05).Compared with the control group,the protein levels of P-IRS1,P-NR1,P-JNK and P-p44/42MAPK were increased by 79.2％,35.1％,47.6 ％,64.3 ％ and 87.6 ％,respectively in diabetic group and 49.4 ％,19.1％,16.5 ％,31.8％ and 39.9％,respectively in insulin-treated diabetes group (all P＜0.001 or 0.05).In comparison with diabetic group,the increased amplitudes of above 4 parameters were decreased by 29.8％,16.0％,31.2％,32.5％ and 47.7％ respectively in the insulin-treated diabetes group (all P＜0.05).Conclusions NMDAR and its downstream signals,such as JNK and p42/44MARK,are involved in the pathogenesis of painful diabetic neuropathy.Although it could not efficiently control the blood glucose level,low-dose insulin treatment may partly inhibit the occurrence of thermal hyperalgesia through inhibiting NMDAR signal pathway.

Application of fractional radiofrequency in cosmetic dermatology / 中华皮肤科杂志

Fractional radiofrequency(FRF)is an aesthetic technique that utilizes electric current emanating from electrode or microneedle arrays to heat the dermis in a fractional pattern, with only little damage to the epidermis. The recovery process following the heat damage involves multiple heat shock proteins, matrix metalloproteinases, cytokines, etc, which can stimulate the proliferation of collagen and elastic fibers in the dermis. It has been applied to treat wrinkles, skin laxity, acne scars and other aesthetic skin problems, and has proved to be a safe and effective cosmetic method for the improvement of atrophic acne scars, inflammatory acnes and postinflammatory erythema. It has few adverse effects, including tolerable pain, transient erythema, edema and mild crusting, so the downtime is short. FRF is more suitable for populations with dark complexions because of low risks of postinflammatory hyper?pigmentation. Recently, it has been used in combination with other devices such as lasers, or been used to assist transdermal drug delivery, and has shown remarkable therapeutic effects and favorable application prospects.

Effect of ginkgolide B on TLR4 and inflammatory protein expression in high glucose treated human umbilical vein endothelial cells / 中国药理学通报

Aim To investigate the effect of ginkgolide B on TLR4 expression in glucose-treated endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs)were stimulated by high concentra-tion of glucose.TLR4,inflammatory protein expression and Akt phosphorylation were analyzed by Western blot.Transcription factor NF-κB nuclear translocation was analyzed by immunofluorescence.Results The expression of TLR4 and PAF receptor was increased in high glucose-treated HUVECs. In contrast, both ginkgolide B and CV3988 dose-dependently decreased TLR4 and PAF receptor expression in high glucose-treated cells,respectively.Ginkgolide B decreased in-flammatory protein ICAM-1 ,VCAM-1 expression.Mo-reover,ginkgolide B potently abolished Akt phospho-rylation and NF-κB p65 nuclear translocation.Conclu-sion Ginkgolide B can reduce TLR4,PAF receptor, ICAM-1 and VCAM-1 expression in high dose of glu-cose-treated HUVECs,the mechanism might be linked to inhibition of Akt phosphorylation and NF-κB activa-tion.

Effects of metformin on expression of AMP-activated protein kinase in rat glomerular mesangial cells / 中华肾脏病杂志

Objective To observe the effects of metformin on expression of Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK),nuclear factor-κB (NF-κB) and transforming growth factor β1 (TGF-β1) in cultured rat glomerular mesangial cells (MCs),and explore its renoprotective mechanisms.Methods MCs were cultured in the medium with normal glucose (group NG,5.6 mmol/L),high glucose (group HG,25mmol/L) and different concentrations of metformin (group M1,M2,M3).After 48 h exposure,the supernatants and MCs were collected.The expression of NF-κB and TGF-β1 mRNA was analyzed by real time-PCR.Total-AMPK,phospho-Thr-172 AMPK (p-AMPK),NF -κB p65 and TGF-β1 were visualized by Western blot.Results The real time-PCR and Western blot result showed MCs could express AMPK,NF-κB and TGF-β1 mRNA and protein.After stimulated by HG,the levels of intracellular NF-κB and TGF-β1 expressions were significantly increased compared with group NG (P ＜ 0.05); The levels of NF-κB and TGF-β1 were significantly decreased in group M1,M2 and group M3 compared with group HG in a dose-dependent manner.After stimulated by HG,the level of intracellular p-AMPK were down-regulated compared with group NG(all P ＜ 0.05); The expression of p-AMPK increased with the rising of metformin concentration,presenting the opposite trend (P ＜ 0.05),while the level of total-AMPK protein was unchanged with exposure to HG or different concentrations of mefformin(P ＞ 0.05).Conclusion Metformin can suppress the expression of NF-κB and TGF-β1 of glomerular MCs induced by HG via AMPK activation,which may partly contribute to its reno-protection.

Effect of ginkgolide B on junctional proteins in oxidized LDL-stimulated human umbilical vein endothelial cells / 中国药理学通报

Aim To investigate the effect of ginkgolide B on junctional proteins in ox-LDL-stimulated human umbilical vein endothelial cells ( HUVECs) . Methods After incubation with ginkgolide B ( 0 . 2 ,0 . 4 ,0 . 6 g · L-1 ) for 1 h, HUVECs were treated with ox-LDL (0. 1 g·L-1 ) for 4 h. The expressions of JAM-A and Cx43 were analyzed with Western blot and immunofluo-rescence. The effect of ginkgolide B on vascular per-meability was analyzed by Transwell experiments. Re-sults JAM-A and Cx43 expressions increased by 22%and 24% in ox-LDL-treated HUVECs, respectively. Whereas ginkgolide B significantly decreased JAM-A and Cx43 expressions. LY294002, a specific inhibitor of PI3K, suppressed JAM-A and Cx43 expressions in ox-LDL-stimulated cells. Ginkgolide B potently re-duced monocyte migration in ox-LDL-treated cells. Conclusion Ginkgolide B significantly suppresses JAM-A and Cx43 expressions, and reduces monocyte migration in ox-LDL-stimulated cells. This demon-strates that ginkgolide B can improve vascular permea-bility. The mechanism might be associated with the in-hibition of PI3K/Akt signaling pathway.

Metformin inhibits nuclear factor-κB activation and inflammatory cytokines expression induced by high glucose via adenosine monophosphate-activated protein kinase activation in rat glomerular mesangial cells in vitro / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The renoprotective mechanisms of adenosine monophosphate (AMP)-activated protein kinase (AMPK) agonist - metformin have not been stated clearly. We hypothesized that metformin may ameliorate inflammation via AMPK interaction with critical inflammatory cytokines. The aim of this study was to observe the effects of metformin on expression of nuclear factor-κB (NF-κB), monocyte chemoattractant protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1) and transforming growth factor-beta 1 (TGF-β1) induced by high glucose (HG) in cultured rat glomerular mesangial cells (MCs).</p><p><b>METHODS</b>MCs were cultured in the medium with normal concentration glucose (group NG, 5.6 mmol/L), high concentration glucose (group HG, 25 mmol/L) and different concentrations of metformin (group M1, M2, M3). After 48-hour exposure, the supernatants and MCs were collected. The expression of NF-κB, MCP-1, ICAM-1, and TGF-β1 mRNA was analyzed by real time polymerase chain reaction. Western blotting was used to detect the expression of AMPK, phospho-Thr-172 AMPK (p-AMPK), NF-κB p65, MCP-1, ICAM-1, and TGF-β1 protein.</p><p><b>RESULTS</b>After stimulated by HG, the expression of NF-κB, MCP-1, ICAM-1, TGF-β1 mRNA and protein of MCs in group HG increased significantly compared with group NG (P < 0.05). Both genes and protein expression of NF-κB, MCP-1, ICAM-1, TGF-β1 of MCs induced by high glucose were markedly reduced after metformin treatment in a dose-dependent manner (P < 0.05). The expression of p-AMPK increased with the rising of metformin concentration, presenting the opposite trend, while the level of total-AMPK protein was unchanged with exposure to HG or metformin. Conlusion Metformin can suppress the expression of NF-κB, MCP-1, ICAM-1 and TGF-β1 of glomerular MCs induced by high glucose via AMPK activation, which may partly contribute to its reno-protection.</p>

The molecular characteristics of heteroresistant vancomycin-intermediate Staphylococcus aureus in China / 中华检验医学杂志

Obiective To investigate the molecular characteristics of heteroresistant vancomycinintermediate Staphylococcus aureus(hVISA)in China and analyze the differences of the molecular characteristics between hVISA and VSSA(vancomycin-susceptible Staphylococcus aureus)isolates.Methods A total of 3 15 non-repetitive MBSA were collected from the national surveillance program in China in 2007.The isolates of hVISA were confirmed by modified population analysis profile-area under the curve(PAP-AUC).The genotypes of agr and SCCmec were determined by multiplex PCR,and spa typing was performed bv PCR and DNA sequencing.The pvl gene was detected bv PCR Results The prevalence of hVISA was 9.5%(30/315).Among 315 MRSA,SCCmec Ⅲ was the most popular type,which was found in 234 isolates(234/315,74.3%),followed by SCCmec Ⅱ,which was identified in 56 isolates (56/315,17.8%).The rate of SCCmec Ⅱ in hVISA(46.7%)was significantly hisher than in VSSA (14.7%,X~2=18.93,P＜0.001).The most prevalent agr type among 315 MRSA was agr 1 accounting for 73.6%(232/315).The agr 2 accounted for 18.7%(59/315),and agr 3 and agr 4 were very rare in clinical isolates.It was different in agr types between the two groups.The rate of agr 2 in hVISA(53.4%)was higher than in VSSA(15.1%).X~2 value was 26.08 and P value was less than 0.001 through X~2 test.There was a statistical significance in the result.There were 4 spa types in hVISA isolates,including t002 (13 isolates),t037(9 isolates),t030(6 isolates),and 1548(2 isolates).The pvl positive MRSA isolates were very low,accounting for 1.6%(5/315).Conclusions The prevalence of hVISA was relatively higher in China.Compared to VSSA,the majority(53.4%)of the hVISA strains were agr 2,which was obviously different from VSSA.hVISA isolates were more diverse by spa typing,

Determination of Fluconazole in Human Plasma by RP-HPLC / 中国药房

OBJECTIVE: To establish RP-HPLC method for the determination of fluconazole in human plasma.METHODS: After liquid-liquid extraction,the plasma sample was analyzed on Diamonsil C18 column with column temperature set at room temperature.The mobile phrase consisted of methanol-0.07% aqueous solution of triethylamine(50∶ 50) with a flow rate of 1mL? min-1.The UV detection wavelength was set at 210nm,and phenacetin was used as the internal standard.RESULTS: The linear range of fluconazole was 0.15～ 10.0mg? L-1(r=0.999 2),with lower quantification limit at 0.15mg? L-1.The intra-day RSD was between 3.37% and 7.48% and the inter-day RSD was between 7.03% and 12.05%;the method recovery was 103.64%(RSD=5.04%),and the average extraction recovery was between 83.81%(RSD=5.77%).CONCLUSION: This method was sensitive,accurate,rapid and reproducible,and suitable for the content determination of fluconazole in plasma.