ABSTRACT
Objective:To explore the pathological differences of surgically resected specimens of advanced esophageal squamous cell carcinoma (ESCC) to different neoadjuvant therapies (neoadjuvant radiochemotherapy and toripalimab combined with neoadjuvant radiochemotherapy).Methods:Thirty patients diagnosed with advanced ESCC who underwent surgical operation after neoadjuvant therapy in Jiangsu Cancer Hospital from October 2020 to September 2021 were included. Among them, 15 patients received neoadjuvant radiochemotherapy (radiochemotherapy group) and 15 patients were treated with toripalimab combined with radiochemotherapy (immunotherapy combined with radiochemotherapy group). Surgically resected specimens were collected. The histopathological features of primary esophageal lesions and the responses of involved lymph nodes were analyzed and compared between two groups.Results:The major pathological response (MPR) rate in the radiochemotherapy group was 10/15, and 14/15 in the immunotherapy combined with radiochemotherapy group ( P=0.17). The pathological complete response (pCR) rate of the primary lesions in the radiochemotherapy group was 7/15, and 10/15 in the immunotherapy combined with radiochemotherapy group ( P=0.46). In the radiochemotherapy group, the incidence rate of tertiary lymphoid structure (TLS) was 7/15, and 12/15 in the immunotherapy combined with radiochemotherapy group ( P=0.02). The incidence rate of necrosis in the radiochemotherapy group was 6/15, and 1/15 in the immunotherapy combined with radiochemotherapy group ( P=0.03). In addition, the incidence rate of foam cell infiltration in the radiochemotherapy group was 6/15, and 13/15 in the immunotherapy combined with radiochemotherapy group ( P=0.01). Furthermore, the pCR rate of involved lymph nodes in the radiochemotherapy group was 7/33, and 11/12 in the immunotherapy combined with radiochemotherapy group ( P<0.001). Conclusion:Compared with the radiochemotherapy group, the incidence of TLS and foam cell infiltration is higher, the incidence of necrosis is lower and clinical efficacy of involved lymph nodes is higher in the immunotherapy combined with radiochemotherapy group, prompting that toripalimab combined with neoadjuvant radiochemotherapy exert higher synergistic immune effect.
ABSTRACT
Human Gasdermin B (GSDMB) gene, as a member of the Gasdermin (GSDM) gene family, may be associated with the development of asthma, tumor and immune system diseases. Recent studies have found that cell pyroptosis can be mediated by GSDMB protein. The N-terminus of GSDMB cleaved by Granzyme A (GZMA), which is secreted by cytotoxic lymphocytes, can directly promote cell pyroptosis. Moreover, GSDMB protein promotes the cleavage of Gasdermin D (GSDMD) by binding to cysteinyl aspartate specific proteinase-4 (caspase-4), thus indirectly promoting cell pyroptosis. This article summarized the progress in the mechanism of GSDMB gene-mediated cell pyroptosis and the related diseases.
ABSTRACT
Objective:To express truncated glycoprotein (Gn, Gn1, Gn2, Gn3, Gc1 and Gc2) of Guertu virus (GTV) in Escherichia coli ( E. coli) cells, and prepare polyclonal antibodies against recombinant proteins Gn-His, Gc1-His and Gc2-His after purification. Methods:Gene fragments encoding Gn, Gn1, Gn2, Gn3, Gc1 and Gc2 of GTV DXM strain were amplified by RT-PCR, and cloned into the prokaryotic expression vector pET-32a (+ ) to construct recombinant expression plasmids. The transformed E. coli BL21(DE3) strains carrying expression plasmids were induced by IPTG to express target proteins, which were identified by SDS-PAGE. Recombinant proteins Gn-His, Gc1-His and Gc2-His were purified by nickel affinity chromatography and detected by Western blot using GTV-positive sheep serum for analysis of their antigenicity. New Zealand white rabbits were immunized with the purified recombinant proteins. The titers and specificity of serum antibodies were analyzed by ELISA. Meanwhile, eukaryotic expression vectors pcDNA3.1-Gn, pcDNA3.1-Gc1/Gc2 were constructed and transfected into mammalian Vero cells to evaluate the binding activity of rabbit polyclonal antibodies by indirect immunofluorescence method. The specific reactivity of serum antibodies to recombinant proteins was detected by Western blot. Results:Restriction enzyme analysis and DNA sequencing confirmed that the recombinant expression vectors of pET-32a-Gn, pET-32a-Gn1/Gn2/Gn3, pET-32a-Gc1/Gc2, pcDNA3.1-Gn and pcDNA3.1-Gc1/Gc2 were constructed successfully. The relative molecular mass ( Mr) of the expressed recombinant proteins Gn-His, Gn1/Gn2/Gn3-His, Gc1/Gc2-His were approximately 63.4×10 3, 37.1×10 3, 31.9×10 3, 30.8×10 3, 40×10 3 and 54.4×10 3, respectively. The recombinant proteins could be recognized by GTV-positive sheep serum. The titers of polyclonal antibodies against GTV Gn, Gc1 and Gc2 were 1∶409 600, 1∶204 800 and 1∶6 400, respectively. Indirect immunofluorescence assay and Western blot showed that the prepared rabbit polyclonal antibodies could specifically react with the proteins expressed in eukaryotic cells and the recombinant proteins. Conclusions:The recombinant GTV glycoproteins Gn-His and Gc1/Gc2-His were efficiently expressed and purified and characterized with good immunity. The prepared polyclonal antibodies had high titers and good specificity. This study provided reference for further studying the biological function and detection methods of GTV glycoproteins and research on vaccines.
ABSTRACT
Objective To express and purify two domains GcⅠand GcⅡof Xinjiang hemorrhagic fever virus (XHFV) glycoprotein, and to study its immunogenicity and the effects on immune response in mice. Methods The prokaryotic expression plasmids of pET28a-GcⅠand pET32a-GcⅡwere constructed and transformed into E. coli BL21, respectively. The expression and purification conditions of rGcⅠand rGcⅡproteins were optimized. The antigenicity of the fusion protein was detected by Western Blot and enzyme-linked immunosorbent assay (ELISA). BALB/c mice were immunized by protein immunization and DNA priming-protein boosting. The mice were randomly divided into 5 groups, including pVAX1-GcⅠ+rGcⅠgroup, pVAX1-GcⅡ+rGcⅡgroup, rGcⅠgroup, rGcⅡgroup and saline group (control group) with 7 mice in each group. The serum antibody titer of mice was detected by indirect ELISA, and the immune effect was evaluated by spleen T lymphocyte proliferation assay and cytokine content determination. Results The fusion proteins rGcⅠand rGcⅡwere purified and obtained, which could react with positive serum of sheep and had good antigenicity. After three immunizations, the IgG levels in the serum of each experimental group were significantly higher than those in the control group (all P<0.001). The serum antibody titers of the experimental groups were reached above 1:12800. Among them, the concentration of Th2 type cytokine interleukin-4 (IL-4) in the spleen cell culture supernatant of rGcⅡ[(79.97±7.47) ng/L] and pVAX1-GcⅡ+rGcⅡgroup [(61.43±9.27) ng/L] was significantly higher than (24.29±3.81) ng/L of the control group, respectively (all P<0.01). The highest mass concentration [(42.46 ±2.60) ng/L] of Th1 type cytokine interferon-γ(IFN-γ) was observed in the pVAX1-GcⅡ+rGcⅡ group, which was significantly higher than (20.33±1.67) ng/L of the control group, and the difference was statistically significant (P<0.001). That showed a significant antigen-specific splenic T lymphocyte proliferation (P<0.001). Conclusions The purified recombinant proteins rGcⅠand rGcⅡhave good immunogenicity, which can make the immune system T lymphocytes tend to Th2 response, and pVAX1-GcⅡ combined with recombinant protein GcⅡ can induce better antigen-specific immune effect. And pVAX1-GcⅡ combined with recombinant protein GcⅡis expected to be used as vaccine candidates for the prevention and control of XHFV.
ABSTRACT
Objective@#To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy.@*Methods@#This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS.@*Results@#There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109).@*Conclusion@#There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.
ABSTRACT
The disease caused by Rickettsia is an important zoonotic disease. Due to the complexity of the ecological environment of Xinjiang, the vectors, host and species of Rickettsia are complex and diverse; its natural foci are widely distributed, so they have brought great harm to local human health and livestock production. With global warming, rapid development of social economy and promotion of foreign economic and trade exchanges in Xinjiang, these may become potential factors for the diversity of Rickettsia vectors and hosts. In recent years, some rickettsial disease such as the onset of Q fever, ehrlichiosis and anaplasmosis, are increased at some areas of Xinjiang, thus, the Rickettsia and rickettsial diseases have been widely concerned. At present, a variety of new Rickettsia species have been detected and isolated from several areas of Xinjiang, and their public health significance have also been paid more and more attention. In this paper, we briefly summarized the research status about reported Rickettsia species identification, media host and popular distribution in the Xinjiang region over the past 20 years, to providing scientific basis for a systematic study and diagnosis, prevention and treatment of Rickettsia and rickettsial disease.
ABSTRACT
Objective To compare the impact of tobacco control curriculum on self-efficacy of tobacco control in different major of public health students, and offer theoretical foundation for improving tobacco control curriculum.Methods We offered tobacco control curriculum for 150 public health students (including students of preventive medicine and public service administration research direction) in medical university of Harbin, and investigated twice general self-efficacy, smoking selfefficacy and tobacco control self-efficacy before and after tobacco control curriculum, then compared intervention effects between two major students by t test.Results There were no differences of tobacco control self-efficacy, smoking self-efficacy and general self-efficacy between two major students, t values were-0.832 (P=0.407), 0.190 (P=0.849) and-0.492 (P=0.624).The tobacco control self-efficacy (t=-3.566, P=0.001) and general self-efficacy (t=-2.237, P=0.028) increased remarkably but smoking self-efficacy (t=-1.102, P=0.273) in preventive medicine students after intervention.And all of three kinds of self-efficacy were not gotten better by intervention in public service administration students.There was no obvious difference of intervention effects between tobacco control self-efficacy and general self-efficacy among preventive medicine students (t=1.541, P=0.125).Conclusions The improvement of preventive medicine students by tobacco control curriculum is better than that of public service administration students.We should further perfect the tobacco control course, and on the one hand, improve the intervention effect of students majoring in course of public health management;on the other hand, strengthen the target effects of the tobacco control curriculum to students' tobacco control self-efficacy and smoking self-efficacy.
ABSTRACT
Objective To study the expression changes of CD54 and CD106 in peripheral blood lymphocyte in patients with congest heart failure. Methods With FCM technique, the levels of CD54 and CD106 in lymphocyte from patients with CHF were measured , and those of patients with hypertension , patients with dilated cardiomyopathy and normal controls were measured at the same time. Cardiac function during heart failure episodes and remission stage was monitored by Color Doppler Echocardiography. Results Levels of CD54 and CD106 were significantly elevated in patients with hypertension , patients with active CHF and hypertension , patients with inactive CHF and hypertension when compared with those of normal controls. Levels of CD54 and CD106 were significantly elevated in patients of dilated cardiomyopathy , patients with active CHF and dilated cardiomyopathy , patients with inactive CHF and dilated cardiomyopathy when compared with those of normal controls. Levels of CD54 and CD106 in patients of CHF were elevated with the degree of CHF. There was significantly negative correlation between LVEF and CD54 of CHF. Conclusions CD54 and CD106 may use as the marker to monitor the progress of CHF.
ABSTRACT
OBJECTIVE: To investigate the pathogenic mechanism of aristolochic acid nephropathy (AAN) by observing the renal tubular injury and the change of the expression of bone morphogenetic protein-7 (BMP-7) mRNA in renal tissue of rats induced by aristolochic acid (AA), an active constituent in Caulis Aristolochiae Manshuriensis (CAM). METHODS: Forty-six male Wistar rats were randomly divided into normal control group (n=20) and AA-treated group (n=26). Rats in AA-treated group were intragastrically administered with AA 20 mg/(kg.d), and rats in control group were treated with equal volume of potable water. At the end of the 4th, 8th and 12th week of administration, the 24 h-urine was collected by metabolic cage for detecting the activity of N-acetyl-beta-D-glucosaminidase (NAG) and the blood samples were obtained from abdominal aorta for detecting serum creatinine (SCr). Pathological change and the degree of injury of the kidneys were observed by microscopy. The expression of proliferative cell nuclear antigen (PCNA) was detected by immunohistochemical method, and mRNA expression of bone morphogenetic protein-7 (BMP-7) in the renal tissue was detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Compared with the normal control group, the activity of NAG and the ratio of SCr vs body weight were markedly increased in rats of the AA-treated group after treatment (P<0.05 and P<0.01). Pathological section of renal tissue showed that most renal tubules had cloudy swelling, and vacuolar degenerating in tubular epithelial cells, with brush border dropping off, and parts of tubular basement membrane were exposed. The degrees of injuries were aggravated depending on treating time. The tubulointerstitial injury (TI) parameter in rats of AA-treated group was higher than that of the normal control group. The positive expression of PCNA was observed in the damaged tubular cells. The proliferation index of PCNA was significantly increased after 4- and 8-week treatment (P<0.01), but was decreased after 12-week treatment (P<0.05). The mRNA expression of BMP-7 was markedly decreased in the AA-treated group compared with the normal control group after 4-week treatment (P<0.05), and decreased with the extension of treatment time. CONCLUSION: AA can induce injury of the renal tubules, impair the cell regeneration, and inhibit the expression of BMP-7 mRNA in renal tissue. This may be one of the pathogenic mechanisms of AAN.
ABSTRACT
Based on the accurate cognition about tumor pathology,relying on observation of clinic cases,combining unsettled problems about tumor,instructed by theory of pathogenesis,the project puts forward the cancerous toxin-rooted and rising and falling of vital qi and pathogen-determined tumor viewpoint.The purpose of this paper is to provide methods and theory for treatment of tumor.
ABSTRACT
Object: To explore the effects and the influent agents of early education on the healthy termed infants. Methods: 52 healthy termed infants voluntary to the exercise center as experimental group were early educated. The other 52 infants as control group were in the same elementary conditions comparing to the experimental group. The experimental group was trained the corresponding projects and examined DQ at the beginning and the 3 rd,6 th,9 th month after training. The control group was only examined DQ at the beginning and after 9 months. Results:①At the 9 th month after training, DQ of the experimental group was improved 21.9?13.6, which was much 7 times higher than that of the control group, which was improved 3?3.0 (P
ABSTRACT
Objective To study the tissue pathology and the expression of related factors during renal tubular-interstitial fibrosis in aristolochic acid (AA) nephropathy in rat. Methods 46 male Wistar rats were randomly divided into 2 groups. The test group consisted of 26 rats which were gavaged with the extract of Caulis Aristolochiae Manshuriensis (CAM) (AA 20mg?kg -1 ?d -1 ); the control group consisted of 20 rats which were given with equal volume of potable water. At the end of 4th, 8th, 12th week, the kidneys of each rat were separately harvested. The HE, PAS and Masson staining were used to analyze the degree of tubular damage and interstitial fibrosis, and immunohistochemical method was applied to assess the protein expression of proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF), transforming growth factor-?_1 (TGF-?_1) in the renal specimens. The mRNA expression of VEGF, endothelin-1 (ET-1), bone morphogenetic protein-7 (BMP-7) in renal tissue were determined by RT-PCR respectively. Results A severe renal tubular-interstitial damage and an early fibrosis were observed at the end of 12th week, and the interstitial fibrotic area was 31.36%. The protein expression of PCNA was increased at 4th week, but down-regulated after 8th week; the expression of TGF-?_1 and VEGF was increased at 4th week, while TGF-?_1 was maintatined on a high level with passage of time, but VEGF decreased gradually. The mRNA expression of VEGF and ET-1 increased notably at 4th week, slightly decreased after 8th week, but maintained at a high level. The BMP-7 declined slowly with the progression of pathological changes, reaching its lowest level at 12th week. Conclusion The mechanism of the rapid progression of fibrosis in AAN might be the renal result of severe impairment of regeneration of epithelial cells, lowering of expression of factors of promoting repair and inhibiting fibrosis, while the expression of factors of promoting fibrosis was maintained at a highlevel.
ABSTRACT
Objective To investigate the effects of aristolochic acid on renal function and tissues morphology in rats.Methods Forty-six male Wistar rats were divided into the normal control group(n=20) and the tested group (n=26) at random.The tested group was treated with the extract of Caulis Aristolochiae Manshuriensis(CAM) (aristolochic acid 20 mg?kg-1?d-1) by gastric gavage,the control group with the same volume of potable water.The body weight of rats was meausured weekly.At the end of 4th,8th,12th weeks,the 24-hour-urine were collected,and blood samples were taken from abdominal artery for detection of indexes of renal function.The pathological changes of renal tissues were also observed.Results The body weight increased slowly and urine volume increased in tested group.The urine albumin level and N-acetyl-?-glucosaminidase (NAG) excretion increased significantly,and the ratio of serum creatinin and body weight increased progressively,indicating that renal function has been impaired.The renal pathological changes under light microscope were as follows:degeneration and necrosis of renal tubule in the early stage and serious tubular structure disorders of focal atrophy and interstitial inchoate fibrosis in the late stage.The changes of ultrastructure of renal tissues under the electronic microscope were as follows:obvious organelle impairment and nucleus variation,interstitial inchoate fibrosis occurring at the 8th week and fibrosis rate arriving 31.36 %at the 12th week.Conclusion Aristolochic acid in CAM has renal toxicity,which is shown as remarkably renal pathological changes,tubular-interstitial injury,interstitial inchoate fibrosis,and cell nuclear variation.
ABSTRACT
90 ). ③For the experimental group at baseline, DQ of the gross movement was the highest (71.5?10.5), while that of the other four areas were similar (fine movement 56.7?10.4, adaptability 59.6?12.5, language 54.3?11.7, social 56.8?14.7).Conclusions:Early intervention had significant effects for the development of children with mental retardation, which medicine has less role.
ABSTRACT
AIM: To observe the role of peroxysome proliferator activated receptor-?(PPAR-?) and the relationship of cyclooxygenase-2(COX-2) and PPAR-? in injury of cultured rat cortical neurons induced by hypoxia/reoxygenation.METHODS: Primary rat cortical neurons were cultured.Experiments include control group,hypoxia/ reoxygenation group and hypoxia/ reoxygenation with PPAR-? agonist group.Cell viability was surveyed by MTT assay.COX-2 protein expression was measured by Western blotting.RESULTS: Neuron viability raised dramatically in hypoxia/reoxygenation with PPAR-? agonist group,compared with hypoxia/reoxygenation group(P