Objective:To study the clinicopathologic characteristics and the prognostic factors of triple negative breast cancer,and to study the correlation between clinicopathologic features and EGFR expression.Methods:Clinical characteristics and prognosis of 200 breast cancer patients diagnosed between 1997 and 2004 were reviewed.The paraffin embedded tissues were selected and tissue microarray blocks were made.Immunohistochemical staining was used to evaluate the expression of ER,PR and HER2 for breast cancer molecular type.EGFR expression was evaluated to analyze its correlation with the clinical characteristics and prognosis.Results:Fourty-two cases(21%)of 200 were triple negative breast cancer(TNBC).There was no significant difference between TNBC and non-TNBC in clinicopathologic parameters such as age,tumor size,clinical stage,and lymph node status.However,there was significant difference in histological category and menostasis status between the two groups(P<0.05).Among TNBC,17 0f 42 died during the follow-up(the overall suivival rate was 59.52%),while 26 0f 158 non-TNBC died(the overall survival rate was 83.54%).EGFR was significantly overexpressed in TNBC(69.05%)and it was associated with lymph node status and histological category.Conclusion:Although the incidence of TNBC is low,it has a poorer prognosis than other subtypes.EGFR is highly expressed in TNBC and may serve as a prognostic index and an important therapatic target for breast cancer.

OBJECTIVEIsobaric tags for relative and absolute quantitation (iTRAQ) combined with mass spectrometry were used to screen differentially expressed plasma proteins in cold stress rats.

METHODSThirty health SPF Wistar rats were randomly divided into cold stress group A and control group B, then A and B were randomly divided into 3 groups (n = 5): A1, A2, A3 and B1, B2, B3. The temperature of room raising was (24.0 +/- 0.1) degrees C, and the cold stress temperature was (4.0 +/- 0.1) degrees C. The rats were treated with different temperatures until 12 h. The abdominal aortic blood was collected with heparin anticoagulation suction tube. Then, the plasma was separated for protein extraction, quantitative, enzymolysis, iTHAQ labeling, scx fractionation and mass spectrometry analysis.

RESULTSTotally, 1085 proteins were identified in the test, 39 differentially expressed proteins were screened, including 29 up-regulated proteins and 10 down-regulated proteins. Three important differentially expressed proteins related to cold stress were screened by bioinfonnatics analysis (Minor histocompatihility protein HA-1, Has-related protein Rap-1b, Integrin beta-1).

CONCLUSIONIn the experiment, the differentially expressed plasma proteins were successfully screened in cold stress rats. iTRAQ technology provided a good platform to screen protein diaguostic markers on cold stress rats, and laid a good foundation for further. study on animal cold stress mechanism.

Animals ; Blood Proteins ; chemistry ; Cold Temperature ; Mass Spectrometry ; Rats ; Rats, Wistar ; Stress, Physiological

Abstract: Objective　To develop a real-time fluorescent quantitative RT-PCR (qRT-PCR) method for qualitative and quantitative Chikungunya virus (CHIKV) analysis. Methods Based on the systematic analysis of the genomic sequences of Chikungunya and its related arboviruses, the specific nucleic acid sequences for Chikungunya virus were screened and identified, and then the primers and TaqMan probe were designed. Meanwhile, the human GAPDH gene was used as an internal reference. The reaction system for qRT-PCR was systematically optimized by L9(34) orthogonal design, and a rapid detection method for Chikungunya by qRT-PCR based on TaqMan probe methods was established. The sensitivity, specificity, reproducibility, and coverage of the established method were analyzed in detail. The standard curve was made, and the absolute quantitative method was established using the cloned nucleic acid fragments as positive samples. Results　A real-time fluorescent quantitative RT-PCR assay was developed for the qualitative and quantitative analysis of Chikungunya virus. The reaction system included Chikungunya virus and reference internal gene specific primers and probe, RT/Taq enzyme mixture, reaction buffer, and negative and positive reference. The established method obtained positive results with the ROSS strain of ECSA subtype, LR2006 strain of IOL branch, 181/25 strain of Asian type and Dongguan 2010 epidemic strains of Chikungunya virus, but there was no cross-reaction with other 18 arboviruses belonging to Flaviviruses, Alphaviruses and Bunyavirus. The minimum detection limit of the established method was 5.80 copies/mL, and a linear relationship was observed between the amount of input plasmid DNA and fluorescence signal value over a range of 5.80×102 copies/mL to 5.80×1010 copies/mL, and the correlation coefficient was 0.999 5. The qRT-PCR amplification efficiency was 91%, and the intra-assay variations and inter-assay variations were 0.01-0.07 and 0.03-0.11, respectively. Conclusions　The TaqMan qRT-PCR method developed in this study can qualitatively and quantitatively detect Chikungunya virus rapidly with specificity and sensitivity, providing a technical method for the prevention and control of this viral disease.

AIM To investigate the effect of Wendan Decoction on nerve injury in a mouse model of sleep disorders and its mechanism.METHODS A mouse model of insomnia was established by the modified multiple platform sleep deprivation method.After successful modeling,the mice were randomly divided into the model group,the estazolam tablet group(0.15 mg/kg)and the low-dose and high-dose Wendan Decoction groups(12.5,50 g/kg),with 6 mice in each group,in contrast to the 6 mice of the control group.After 7 days of drug intervention,the mice had their changes of cerebral cortex,hippocampal CA1 area and hypothalamus observed by HE staining;their neuronal damage observed by Nissl staining;their levels of neurofilament light chain(NEFL),neuron-specific enolase(NSE),S100 calcium-binding protein B(S100B),tumor necrosis factor(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)in brain tissue and serum detected by ELISA;their cerebral expression of glial fibrillary acidic protein(GFAP)detected by immunohistochemical method;and their cerebral expressions of GFAP,phosphorylated IκB kinase β(p-IKKβ)and phosphorylated nuclear transcription factor-κB(p-NF-κB)detected by Western blot.RESULTS Compared with the model group,the high-dose Wendan Decoction group displayed increased number of neurons,complete and neatly arranged structure;decreased number of neurons with nuclear shrinkage and deformation;increased Nissl bodies,decreased levels of NEFL,NSE,S100B,TNF-α,IL-6 and IL-1β in serum and brain tissue(P＜0.01);decreased cerebral expression of GFAP(P＜0.01);and decreased phosphorylation levels of cerebral p-IKKβ and p-NF-κB(P＜0.01).CONCLUSION Wendan Decoction can reduce the nerve damage and the expression of proinflammatory mediator in sleep disorders mice,and the mechanism may be related to the inhibited activation of IKKβ/NF-κB pathway.

Objective To investigate the changes and clinical significance of peripheral blood mucosal-associated invariant T(MAIT)cells in children with influenza.Methods Children with influenza who received treatment in the outpatient and inpatient departments of a children's hospital from January to May 2023 were selected and divided into the common type group and the severe type group.Healthy children who underwent physical examination in this hospital during the same period were selected as the healthy control group.Within 24 hours after admission,children's venous blood was drawn for testing;ratios of MAIT cells(CD3+CD161+TCRVα7.2+cells)and MAIT cells expressing PD-1,CD69,perforin,and CD107 α were tested by flow cytometry,respectively.Differences among all the groups were compared.Results Compared with the control group,the proportion of peripheral blood MAIT cells in children with common and severe influenza gradually decreased,while the proportion of CD69-ex-pressing and perforin-positive MAIT cells increased gradually.Differences were statistically significant(all P＜0.05).There was no statistically significant difference in the proportion of MAIT cells expressing CD107(P＞0.05).The proportion of PD-1 positive MAIT cells increased(P＜0.05),but there was no statistically significant difference be-tween the common type and severe type groups(P＞0.05).Conclusion The decrease of peripheral blood MAIT cells accompanied with immune activation plays a role in the pathogenesis of influenza.

The qualitative and quantitative analysis on traditional Chinese medicine and formula components can be made by chemical and instrumental analysis methods. Of both, the instrumental analysis methods play a dominant role, including HPLC, HPLC-MS, HPLC-NMR, GC, GC-MS, biochemical and biological effect. But because traditional Chinese medicines and formula have complicated components, chemical methods are so unspecific that they shall be used less or with caution. While instrumental analysis methods are so specific that they are appropriate for analyzing complicated single component. The analysis techniques for multiple components of traditional Chinese medicines and formula focus on fingerprints, but all of these analysis techniques are limited by the pre-requisite of separation and the lack of general-purpose detectors and therefore being hard to realize the determination of all components of traditional Chinese medicines and formula. In the natural world, however, organisms identify native and alien components through specificity and non-specificity of clusters decided by antigens and antibodies. For example, components of traditional Chinese medicines are directly or indirectly synthesized into antigens and injected into animals, in order to generate specific antibodies and then collect cross reaction information of these components to specific antibodies. As for components without cross reaction, their contents shall be directly read out on the basis of the inhibition rate curve of competitive reaction for specificity of antigens and antibodies. Besides, a cross inhibition rate matrix shall be established first, and them a multiple regression linear equation between cross component concentration or concentration logarithm and inhibition rate by labeling the immunity competitive reaction between antibodies and haptens of traditional Chinese medicine and compound components, and then solved to obtain concentration of each component. The two results are combined to establish the synthetic immunity chip method for traditional Chinese medicine and formula components.
Animals ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; High-Throughput Screening Assays ; methods ; Humans ; Immunohistochemistry ; methods ; Vaccines, Synthetic ; chemistry

Objective To compare the spiral CT and pathological findings of sub-centimetre pulmonary ground glass nodules(GGNs), and to improve the aaccuracy in early diagnosis.Methods The CT findings and pathologic features of 30 patients with sub-centimetre pulmo-nary GGNs in our hospital from May 2013 to June 2016 were reviewed retrospectively.The patients were divided into 3 groups according to their pathological classification,namely the benign group,the preinvasive group and the invasive group.The shape,size,margin,vacuole sign, pleural indentation,the relations with bronchi and blood vessels of the pulmonary GGNs were analyzed.Results There was no significant difference in shape,maximum diameter and margin between the benign group and the other two groups(P>0.05).Adjacent vessels changes of the lesion showed significant differences between benign group and preinvasive or invasive groups(P<0.05).And there were significant differences in size,pleural indentation,vacuole sign and adjacent vessels changes between the preinvasive group and the invasive group(P<0.05).Conclusion The comprehensive analysis of lesions size,vacuole sign,pleural indentation and adjacent vessels changes can be helpful to improve the aaccuracy of differential diagnosis on sub-centimetre pulmonary ground glass nodules.

OBJECTIVETo explore the risk factors for early recurrence (recurrence within 2 years) of proximal gastric cancer after radical resection.

METHODSThe clinical data of 367 proximal gastric cancer patients who underwent radical resection in the Cancer Institute and Hospital of Tianjin Medical University between January 2000 and May 2006 were reviewed. Among them, there are 71 patients (19.3%) with early recurrence. Univariate analysis and multivariate analysis were applied to investigate risk factors for early recurrence.

RESULTSUnivariate analysis showed that Borrmann type (P<0.01), histology type (P<0.01), depth of invasion (P<0.05), negative lymph nodes count (P<0.05) were risk factors for early recurrence of proximal gastric. On multivariate analysis, histology type (P<0.05), depth of invasion (P<0.05), negative lymph nodes counts (P<0.05) were independent risk factors for early recurrence of proximal gastric cancer. Negative lymph nodes in early recurrence patients were 8.4 ± 7.2, which were significantly less as compared to patients without early recurrence (10.7 ± 8.7) (P<0.05).

CONCLUSIONFor T3 proximal gastric adenosquamous cancer, extended resection and lymphadenectomy should be considered. Intraoperative or postoperative adjuvant treatment should be administered as routine.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; pathology ; Postoperative Period ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; pathology ; surgery

OBJECTIVETo investigate the changes in phosphorylated JAK2 and STAT3 protein expression of and cell apoptosis following focal cerebral ischemia-reperfusion injury in rats.

METHODSA rat models of focal cerebral ischemia-reperfusion injury was established by middle cerebral artery occlusion using modified filament method. Immunohistochemistry and Western blot analysis were used to detect the expression of P-JAK2 and P-STAT3 proteins, and TUNEL assay was employed to examine the cell apoptosis.

RESULTSP-JAK2 and P-STAT3 protein expression increased significantly after cerebral ischemia-reperfusion injury in rats. The immunoreactivity was prominent in the peripheral of the ischemic region and reached the peak level at 24 h of reperfusion, followed by slight decrement. The apoptotic cells increased obviously after cerebral ischemia-reperfusion injury, also reaching the peak level at 24 h of reperfusion.

CONCLUSIONThe expression of phosphorylated JAK2 and STAT3 may be involved in the ischemic cellular events including apoptosis. JAK2/STAT3 signaling pathway plays a role in the pathophysiological process of cerebral ischemia/reperfusion cell injury and repair.

Animals ; Apoptosis ; Blotting, Western ; Immunohistochemistry ; In Situ Nick-End Labeling ; Infarction, Middle Cerebral Artery ; physiopathology ; Janus Kinase 2 ; metabolism ; Male ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; genetics ; physiopathology ; STAT3 Transcription Factor ; metabolism

OBJECTIVETo explore the clinicopathological characteristics and prognostic factors of primary gastric lymphoma (PGL).

METHODSThe clinical data of 204 patients with PGL was reviewed and analyzed. There were 106 males and 98 females, their age were 19 to 85 years (average age was 53.7 years). The Focal areas included gastric fundus lesions 41 cases (20.1%), stomach body lesions 127 cases (62.3%), distal gastric lesions 105 cases (51.5%), cardia lesions 13 cases (6.4%), duodenal bulb lesion 1 cases (0.5%). The clinical characteristics and the outcomes in patients with influence were analysed.

RESULTSIn 204 PGL patients, the most common complaints were abdominal pain (62.3%) and weight loss (52.9%). Most of the PGL patients appeared ulcerative (76.0%) and results showed that 62.7% patients involved single location. As to the factors of cellulate grading and pathological characteristics, most patients (87.7%) show low-grade or intermediate-grade lymphoma, Musshoff stages I and II (74.0%). In 186 patients with complete follow-up data, survival rates of 1-, 3- and 5-year were 75.8%, 63.4% and 60.2% respectively. The median overall survival time was 50.0 months. In univariate survival analysis, age (χ(2) = 5.030), level of LDH (χ(2) = 40.084), cellulate grading (χ(2) = 35.238), Musshoff stage (χ(2) = 71.601), tumor diameter (χ(2) = 12.018) and option of managements (χ(2) = 14.140) were obviously correlated with the prognosis (all P < 0.05). Musshoff stage (RR = 2.230, 95%CI: 1.372 - 3.625) and cellulate grading (RR = 1.892, 95%CI: 1.010 - 3.543) were independent prognostic factors in multivariable analysis (both P < 0.05). There was no prognostic difference between surgery and chemotherapy in stage I and II (χ(2) = 1.223, P = 0.542).

CONCLUSIONSMusshoff stage and grade malignancy are independent prognostic factors. For patients with stage I and II, surgical resection is not the first-choice for clinical therapy.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymphoma, Non-Hodgkin ; diagnosis ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; diagnosis ; pathology ; Young Adult