The Effect of Doxycycline on the Expression of MMP-2 and MMP-9 in Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in multiple myeloma (MM) patients, and analyze the effect of doxycycline (DOX) on the expression of MMP-2 and MMP-9 in MM cells.@*METHODS@#The peripheral blood and bone marrow samples of MM patients were collected, and the patients were divided into three groups: newly diagnosed group, remission group and relapsed/refractory group, while the peripheral blood samples of 34 health people and the bone marrow samples of 17 IDA patients were selected as normal control and control group. The levels of MMP-2 and MMP-9 were detected by ELISA. The protein levels of MMP-2 and MMP-9 in H929 cells treated by different concentrations of DOX were analyzed by Western blot. After H929 cells was treated by Akt inhibitor MK-2206 2HCl in combination with DOX, Western blot was used to detect the levels of MMP-2 and MMP-9.@*RESULTS@#The levels of MMP-2 and MMP-9 in newly diagnosed MM patients were higher than those in control (P<0.05), while for the patients in the remission group were decreased, but still higher than those in control. The levels of MMP-2 and MMP-9 were increased again for the patients in relapsed/refractory group, and showed no significant difference as compared with those in newly diagnosed group. The levels of MMP-2 and MMP-9 could be inhibited by 10 mg/L and 15 mg/L DOX treated by H929 cell. The protein levels of MMP-2 and MMP-9 showed no altered in H929 cells treated by 5 nmol/L MK-2206 2HCl alone. DOX exerted more profound inhibitory effect to MMP-2 and MMP-9 expression in H929 cells when Akt inhibitor MK-2206 2HCl was combined with DOX.@*CONCLUSION@#The levels of MMP-2 and MMP-9 are increased in MM patients and related to the disease status of MM. DOX can inhibit the expression of MMP-2 and MMP-9 in MM cells, and antagonizing its activation of Akt signaling pathway can further enhance the inhibitory effect.

Effect of Shugan Yishen Prescription on Tamoxifen-resistant Cell Line from Exosome-mediated Crosstalk in Breast Cancer Microenvironment / 中国实验方剂学杂志

Objective:To explore the effect of Shugan Yishen prescription（SGYS） on the tamoxifen （TAM） -resistant cell line LCC9 by the intervention of exosome-mediated crosstalk in the breast cancer microenvironment. Method:Four groups of serum were set up， specifically， a blank group， a TAM （2 mg·kg^-1·d^-1） group，an SGYS（113.2 g·kg^-1·d^-1） group，and a combination group. The exosomes of LCC9 cells were extracted by ultracentrifugation and identified by transmission electron microscopy （TEM），nanoparticle tracking analysis （NTA）， and Western blot. Then the collected LCC9 exosomes （LCC9-EXO） were co-cultured with bone marrow mesenchymal stem cells（BMMSCs），and 10% of the above four groups of serum were added to the co-culture system. After 48 hours of co-culture，the exosomes of BMMSCs （BMMSCs-EXO） were extracted and incubated with LCC9 cells. Fluorescence microscope was used to observe the uptake of exosomes by cells. Cell counting kit-8 （CCK-8） assay，flow cytometry， and Transwell assay were used to detect the effects of drug-containing serum in the four groups on the proliferation，apoptosis， and migration of LCC9 cells. Western blot was used to detect the protein expression of CD24，CD44，human epidermal growth factor 2（HER2）， and estrogen receptor <italic>α</italic> （ER<italic>α</italic>） in each group. Result:Fluorescence microscope observed that LCC9-EXO could be taken up by BMMSCs，and BMMSCs-EXO could be taken up by LCC9 cells. CCK-8 assay revealed that compared with the TAM group，the SGYS group and the combination group showed reduced cell proliferation ability at each period （<italic>P</italic><0.05），especially the combination group，but no statistically significant difference between the SGYS group and the combination group was observed （<italic>P</italic><0.05）. Flow cytometry revealed that compared with the TAM group，the SGYS group and the combination group showed increased levels of apoptosis （<italic>P</italic><0.05）. Transwell assay revealed that compared with the TAM group，the SGYS group and the combination group showed decreased cell migration ability （<italic>P</italic><0.05）. Western blot revealed that compared with the TAM group，the SGYS group and the combination group showed up-regulated expression of ERα and CD24（<italic>P</italic><0.05），and down-regulated expression of HER2 and CD44 （<italic>P</italic><0.05）. The effect of the combination group on protein expression was superior to that of the SGYS group （<italic>P</italic><0.05）. Conclusion:SGYS reverses the TAM resistance of LCC9 cells by interfering with the crosstalk between BMMSCs-EXO and LCC9-EXO.

Regulatory Effect of Huadu Sanyinfang on PI3K/Akt/NF-κB Pathway in Triple-negative Breast Cancer Patients with Qi-deficiency Constitution： An Exploration Based on Differential Expression of miRNA / 中国实验方剂学杂志

Objective:To explore the regulatory effect of Huadu Sanyinfang on phosphatidylinositol 3 kinase（PI3K）/protein kinase B（Akt）/transcription factor nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） in triple-negative breast cancer （TNBC） patients with qi-deficiency constitution based on the differential expression of miRNA. Method:Based on previous research results， this study conducted the bioinformatics analysis to predict the target genes responsible for regulating the differential expression of miRNA between patients with qi-deficiency constitution and those with moderate constitution， which were intersected with TNBC target genes. The resulting intersection targets were then subjected to Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis and protein-protein interaction （PPI） network analysis to obtain the key pathways and target genes for differentially expressed miRNA in regulating TNBC. TNBC patients with Qi-deficiency constitution were treated with Huadu Sanyinfang for three years after they completed the standard Western medical treatment. The peripheral blood of the patients was sampled before and after medication for detecting gene expression in the key pathways. Result:The comparison between patients with Qi-deficiency constitution and those with moderate constitution revealed 49 differentially expressed miRNAs （16 up-regulated and 33 down-regulated）， which regulated 1 445 TNBC target genes. As demonstrated by PPI and KEGG pathway enrichment analysis， the key genes were mainly tumor protein p53 （TP53）， Akt1， epidermal growth factor receptor （EGFR）， mitogen-activated protein kinase 3 （MAPK3）， vascular endothelial growth factor A （VEGRA）， and tumor necrosis factor （TNF）. The key pathways included PI3K/Akt， MAPK， and RAS signaling pathways. A total of 11 TNBC patients with qi-deficiency constitution were enrolled. Compared with the situations before treatment， the expression levels of p105 subunit of NF-κB （NF-<italic>κ</italic>B1） and Akt1 in the PI3K/Akt signaling pathway were down-regulated after medication， while the levels of catalytic subunit alpha of PI3K （PIK3CA） and B-cell lymphoma-xL （Bcl-xL） were up-regulated. The differences in NF-<italic>κ</italic>B1 and Akt1 expression were statistically significant. Conclusion:Huadu Sanyinfang is able to affect the gene expression of PI3K/Akt/NF-<italic>κ</italic>B signaling pathway in TNBC patients with Qi-deficiency constitution. Specifically， it down-regulates NF-<italic>κ</italic>B1 and Akt1 expression and up-regulates PIK3CA and Bcl-xL.

Discussion on TCM Syndrome Differentiation and Treatment of Triple Negative Breast Cancer / 中国中医药信息杂志

Triple negative breast cancer (TNBC) is a special subtype of breast cancer which lacks expression of the estrogen receptor, progesterone receptor and HER-2. The treatment of TNBC remains a challenge, due to its high heterogeneity, aggressive pattern and insensitivity to endocrine, so the HER-2-directed therapy is ineffective and prognosis is poor. TCM treatment has certain advantages in treating TNBC. The complementation of traditional Chinese and Western medicine should be paid attention to in the process of syndrome differentiation. Differential diagnosis of disease in Western medicine should correspond to treatment based on syndrome differentiation and the concept of wholism in TCM. For different stages of the disease, it is also important to emphasize the relations between supporting the healthy energy and eliminating the evil factors, and the whole and the part before the prescription to analyze and grasp of TCM constitution, to exert the superiority of TCM in treating TNBC.

Comparison of the Predictive Values of Eight Staging Systems for Primary Liver Cancer in Prognosis of Combined Hepatocellular-cholangiocellular Carcinoma Patients after Surgery / 中国医学科学院学报

<p><b>OBJECTIVE</b>To compare the predictive values of eight staging systems for primary liver cancer in the prognosis of combined hepatocellular-cholangiocellular carcinoma (cHCC-CC) patients after surgery.</p><p><b>METHODS</b>The clinical data of 54 cHCC-CC patients who underwent hepatectomy or liver transplantation from May 2005 to Augest 2013 in Chinese PLA General Hospital were collected. We evaluated the prognostic value of the Okuda staging system, Cancer of the Liver Italian Program (CLIP) score, French staging system, Barcelona Clinic Liver Cancer (BCLC) staging system, 7th edition of tumour-node-metastasis (TNM) staging system for hepatocellular carcinoma and intrahepatic cholangiocarcinoma (ICC), Japan Integrated Staging (JIS) score, and Chinese University Prognostic Index. The distribution, Kaplan-Meier method, Log-rank test, and area under a receiver operating characteristic curve were used to compare the prognosis-predicting ability of these different staging systems in 54 cHCC-CC patients after surgery.</p><p><b>RESULTS</b>The TNM staging system for ICC and JIS score had a better distribution of cases. The 12-and 24-month survivals of the entire cohort were 65.5% and 56.3%, respectively. A Log-rank test showed that there was a significant difference existing in the cumulative survival rates of different stage patients when using TNM staging system for ICC (stage 1 vs. stage 2, P=0.012; stage 2 vs. stage 3-4, P=0.002), Okuda staging system (stage 1 vs. stage 2, P=0.025), and French staging system (stage A and stage B, P=0.045). The 12-and 24-month area under curve of TNM staging system for ICC, BCLC staging system, JIS score, and CLIP score were 0.836 and 0.847, 0.744 and 0.780, 0.723 and 0.764, and 0.710 and 0.786, respectively.</p><p><b>CONCLUSION</b>The 7th edition of TNM staging system for ICC has superior prognostic value to other seven staging systems in cHCC-CC patients undergoing surgical treatment.</p>

Apoptosis and its mechanisms of spleen CD4⁺ CD25⁺ regulatory T cells in severe aplastic anemia mouse model / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the apoptosis and its mechanisms of spleen CD4⁺ CD25⁺ regulatory T (Treg) cells in severe aplastic anemia (SAA) mouse model induced by interferon (IFN-γ) in combination with busulphan (BU).</p><p><b>METHODS</b>The BALB/c female mice SAA model was induced by intraperitoneal injection with IFN-γ and intragastric administration with BU (combined group, n=16), with BU group (n=16), IFN-γ group (n=16) and normal group (n=16) as controls. Spleen Treg cells were purified by using of immunomagnetic beads. Apoptosis was detected by flow cytometry. AKt and TGF-β expression was measured by Western blot.</p><p><b>RESULTS</b>Apoptosis of spleen Treg cells in combined group [(33.21±0.65)%] was significantly higher than that in BU group [(21.58±0.64)%], IFN-γ group [(17.62±1.05)%], and control[(27.38±1.89)%] (P<0.05). A significantly lower expression of AKt and TGF-β protein was also seen in combined group (0.30±0.05 and 0.17±0.05), as compared to the other three groups (P<0.05).</p><p><b>CONCLUSION</b>Excessive apoptosis of Treg cells was found in SAA mouse model, which may be a cause of Treg cells decrease in patients with AA. The down-regulated expression of AKt and TGF-β could play a role in increased apoptosis of Treg cells. Our data may provide a new treatment strategy in AA.</p>

A meta-analysis of the long-term effects of chronic pancreatitis surgical treatments: duodenum-preserving pancreatic head resection versus pancreatoduodenectomy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Surgery is regarded as the most effective treatment to relieve pain and reduce complications in chronic pancreatitis (CP). Two major strategies exist: duodenum-preserving pancreatic head resection (DPPHR) and pancreatoduodenectomy (PD). Many studies suggest that DPPHR offers advantages during surgery and in the short-term; however, the long-term effects have not been thoroughly investigated. We analyzed the long-term outcomes of DPPHR and PD, over follow-up times of at least 1 year, to determine the optimal surgical treatment for CP.</p><p><b>METHODS</b>We systemically reviewed all CP surgical treatment reports, and only included randomized controlled trials (RCT) comparing DPPHR and PD, excluding unqualified studies using several pre-specified criteria. When multiple publications of a single trial were found, the most comprehensive current data were selected. Characteristics of the study populations and long-term postoperative outcome parameters were collected. The quality of the studies and data was analyzed using RevMan 4.2 software.</p><p><b>RESULTS</b>Five trials were qualified for meta-analysis, with 261 participants in total (114 in the DPPHR group and 147 in the PD group). There were no significant differences in the age, gender, or indications for surgery of each group. At the mean of 5.7-year (1 - 14 years) follow-up examination, DPPHR and PD resulted in equally effective pain relief, exocrine and endocrine function, and similar mortality rates (P > 0.05); however, DPPHR patients had improved global quality of life and weight gain, and reduced diarrhea and fatigue (P < 0.05).</p><p><b>CONCLUSION</b>DPPHR and PD result in equal pain relief, mortality, and pancreatic function; however, DPPHR provides superior long-term outcomes.</p>

Application of persistent methylene blue dyeing method for anatomic hepatectomy of hepatocellular carcinoma with bile duct tumor thrombi / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the role of anatomic hepatectomy of hepatocellular carcinoma with bile duct tumor thrombi by application of persistent methylene blue dyeing method.</p><p><b>METHODS</b>From January 2009 to February 2011, 11 hepatocellular carcinoma patients with bile duct tumor thrombi underwent anatomic hepatectomy with removal of the biliary tumor thrombus. There were 10 male and 1 female patients. The average age was 49 years (ranging from 31 to 67 years). The initial symptom of 9 out of the 11 patients was jaundice. After anatomy and ligation of Glissonean pedicle of pre-resection segment, methylene blue was injected into its far-end portal vein in order to dye the segment.</p><p><b>RESULTS</b>Persistent methylene blue dyeing method was successful in all patients. Primary foci were found in all patients. Hepatectomy were performed, including 4 patients of segmentectomy, 3 patients of subsegmentectomy, 2 patients of hemihepatectomy, and 2 patients of hepatic sectionectomy. The mean operation time and blood loss was 137 minutes and 246 ml respectively. Severe complications such as liver function failure and sub-diaphragm abscess was avoided in all patients. No perioperative death. Post-operation radiotherapy was performed on 2 patients . Over a mean follow-up time of 14.6 months, liver cancer recurrence occurred in 2 patients, abdomen seeding metastasis in 1 patient, bile duct tumor thrombi recurrence in 1 case, and 2 patients died.</p><p><b>CONCLUSIONS</b>Anatomic hepatectomy of hepatocellular carcinoma with bile duct tumor thrombi by application of persistent methylene blue dyeing method can make resection more precise and improve curative effect.</p>

Clinical related factors of portal vein tumor thrombosis in patients with hepatocellular carcinoma: a logistic regression analysis / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the related factors of portal vein tumor thrombosis (PVTT) in patients with HCC.</p><p><b>METHODS</b>A total number of 234 patients with hepatocellular carcinoma (HCC) were included in this retrospective study. Uni-variate and multi-variate logistic regression analysis were employed to analyze the association between PVTT and 18 routine clinical parameters.</p><p><b>RESULTS</b>Among the 234 patients with HCC, 15% of patients (35/235) had PVTT. Univariate logistic regression analysis revealed significant association of age (P = 0.016), gamma glutamyl transferase (GGT, P = 0.003), number of segmental invasion (P = 0.007), microvascular invasion (P < 0.01), segment location of S2 (P = 0.001), S3 (P = 0.000), S4 (P = 0.004) and S6 (P = 0.016). Multivariate analysis shows potential significant predictors of PVTT in HCC were age (RR: 0.373; 95% CI: 0.146-0.954; P = 0.040), the tumor location of S3 (RR: 4.625; 95% CI: 1.916-11. 165;P = 0.001), GGT (RR: 4.091; 95% CI: 1.448-11.553; P = 0.008) and microvascular invasion (RR: 20.912; 95% CI: 4.745-92.172; P < 0.01).</p><p><b>CONCLUSIONS</b>PVTT occurred more commonly in the younger (< 50 years old), and those with high level of GGT, segment location of S3 and microvascular invasion.</p>

Impact of surgical operation-related factors on long-term survival of patients with hepatocellular carcinoma after hepatectomy / 中国医学科学院学报

<p><b>OBJECTIVE</b>To evaluate the prognostic value of surgical operation-related factors in patients with hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The clinical data of 234 patients after hepatic resection (214 men and 20 women) were retrospectively studied. Univariate and multivariate COX regression analyses were performed for surgical operation-related prognostic factors including age, gender, intraoperative blood loss, iatrogenic tumour rupture, transfusion, operation duration, hepatectomy extent, Pringle manoeuvre, with or without devarscularization, and complications (e.g. postoperative ascites, biliary leakage, incision infection, and pleural effusion). Kaplan-Meier and log-rank tests were used to compare survival rates. Kendall's tau bivariate analyses were used to examine the correlations of these surgical operation-related factors.</p><p><b>RESULTS</b>Univariate COX regression analysis revealed that iatrogenic blood loss (chi2 = 19.721, P < 0.001), transfusion (chi2 = 7.769, P = 0.005), tumour rupture (chi2 = 6.401, P = 0.011), operation duration (chi2 = 4.793, P = 0.029), and postoperative ascites (chi2 = 4.452, P = 0.035) were statistically significant predictors in patients with HCC after hepatic resection. Multivariate COX regression analysis revealed that pathological factors, such as blood loss (RR: 2.138, 95% CI: 1.556-2.939), tumour rupture (RR: 2.260, 95% CI: 1.182-4.321), and postoperative ascites (RR: 1.648, 95% CI: 1.088-2.469), independently influenced the HCC prognosis. Blood loss correlated with transfusion (Kendall's tau = 0.416, P < 0.001). There was no correlation between hepatectomy extent and blood loss (Kendall's tau = 0.057, P = 0.383), while transfusion closely correlated with the hepatectomy extent (Kendall's tau = 0.185, P = 0.004). The postoperative ascites closely correlated with Child classification (Kendall's tau = 0.151, P = 0.024).</p><p><b>CONCLUSIONS</b>The long-term survival of patients with HCC after hepatectomy may be improved by avoiding blood loss and iatrogenic tumour rupture. The indications of blood transfusion may not be strictly obeyed in some severe cases. Child class B and C cirrhotic patients may experience postoperative ascites and a worse prognosis, and therefore may be candidates for liver transplantation.</p>