Diffuse large B-cell lymphoma successfully treated with amplified natural killer therapy alone: A case report

BACKGROUND The prognosis of patients with advanced diffuse large B-cell lymphoma(DLBCL)is poor,with a 5-year survival rate of approximately 50%.The mainstay of treatment is multidrug combination chemotherapy,which has been associated with serious side effects.Amplified natural killer(ANK)cell therapy amplifies and activates natural killer(NK)cells to attack only malignant tumors.As ANK cells attack programmed death ligand 1(PD-L1)-positive tumor cells,ANK therapy is considered effective against adult T-cell lymphoma and malignant lymphoma.CASE SUMMARY Herein,we report a case of an older patient with advanced DLBCL who was successfully treated with ANK immunotherapy.A 91-year-old female visited our hospital with sudden swelling of the right axillary lymph node in April 2022.The patient was diagnosed with stage II disease,given the absence of splenic involvement or contralateral lymphadenopathy.ANK therapy was administered.Six rounds of lymphocyte sampling were performed on July 28,2022.To reduce the occurrence of side effects,the six samples were diluted by half to obtain 12 samples.Cultured NK cells were administered twice weekly.The treatment efficacy was evaluated by performing computed tomography and serological tests every 1 or 2 mo.The treatment suppressed lesion growth,and the antitumor effect persisted for several months.The patient experienced mild side effects.PD-L1 immunostaining was positive,indicating that the treatment was highly effective.CONCLUSION ANK therapy can be used as a first-line treatment for malignant lymphoma;the PD-L1 positivity rate can predict treatment efficacy.

Liver metastasis is the only independent prognostic factor in AFP-producing gastric cancer

AIM:To investigate differences between common gastric cancer andα-fetoprotein(AFP)-producing gastric cancer according to the presence or absence of liver metastasis.METHODS:Between 1997 and 2011,1299 patients underwent gastrectomy for gastric cancer(GC)at our institute and their hospital records were reviewed retrospectively.Patients were immunohistochemically divided into two groups:23 patients(1.8%)with AFPproducing GC and 1276 patients(98.2%)without it.RESULTS:AFP-producing GC patients had a significantly higher incidence of deeper tumors,venous invasion,lymphatic invasion,lymph node metastasis,and liver metastasis and a poorer prognosis(P<0.005)than those without AFP-producing GC.However,multi-variate analysis revealed that AFP-positivity was not an independent prognostic factor.The prognosis of AFPproducing GC was similar to that of AFP-non producing GC according to the presence or absence of liver metastasis.Concerning recurrence,47.8%of patients(11/23)with AFP-producing GC and 20.0%of patients(256/1276)without AFP-producing GC exhibited recurrence.Liver metastasis[90.9%(10/11)]was the most prevalent in AFP-producing GC patients.Multivariate analysis revealed that liver metastasis was the only independent prognostic factor in AFP-producing GC(HR=17.6,95%CI:2.1-147.1;P=0.0081).CONCLUSION:AFP-producing GC is similar to common GC without liver metastasis,which should be specifically targeted in an effort to improve the prognosis of AFP-producing GC patients.

Interrelationship between chromosome 8 aneuploidy,C-MYC amplification and increased expression in individuals from northern Brazil with gastric adenocarcinoma

AIM: To investigate chromosome 8 numerical aberra- tions, C-MYC oncogene alterations and its expression in gastric cancer and to correlate these findings with histo- pathological characteristics of gastric tumors. METHODS: Specimens were collected surgically from seven patients with gastric adenocarcinomas. Immu- nostaining for C-MYC and dual-color fluorescence in situ hybridization (FISH) for C-MYC gene and chromosome 8 centromere were performed. RESULTS: All the cases showed chromosome 8 aneu- ploidy and C-MYC amplification, in both the diffuse and intestinal histopathological types of Lauren. No significant difference (P < 0.05) was observed between the level ofchromosome 8 ploidy and the site, stage or histological type of the adenocarcinomas. C-MYC high amplification, like homogeneously stained regions (HSRs) and double minutes (DMs), was observed only in the intestinal-type. Structural rearrangement of C-MYC, like translocation, was observed only in the diffuse type. Regarding C-MYC gene, a significant difference (P < 0.05) was observed between the two histological types. The C-MYC protein was expressed in all the studied cases. In the intestinal- type the C-MYC immunoreactivity was localized only in the nucleus and in the diffuse type in the nucleus and cytoplasm. CONCLUSION: Distinct patterns of alterations between intestinal and diffuse types of gastric tumors support the hypothesis that these types follow different genetic path- ways.

Decreased histone H2B monoubiquitination in malignant gastric carcinoma

AIM:To investigate H2B monoubiquitination(uH2B)and H3K4 di-and tri-methylation(H3K4-2me,H3K4-3me)levels and their clinical significance in gastric cancer(GC).METHODS:Immunohistochemistry(IGC)was used to detect the differential levels of uH2B,H3K4-2me and H3K4-3me modifications in GC specimens from chemo/radiotherapy-na ve patients who underwent potentially curative surgical resection(n=159)and in a random sampling of non-tumor gastric epithelium specimens(normal controls,n=20).The immunohistochemistry(IHC)-detected modifications were classified as negative,low-level,or high-level using a dual-rated(staining intensity and percentage of positively-stained cells)semi-quantitative method.The relationships between uH2B modification levels and clinicopathological parameters of GC were assessed by a Wilcoxon rank sum test(pairwise comparisons)and the Kruskal-Wallis H test(multiple comparisons).The correlation between uH2B modification and survival was estimated by Kaplan-Meier analysis,and the role of uH2B as an independent prognostic factor for survival was assessed by multivariate Cox regression analysis.RESULTS:The presence and level of H3K4-2me and H3K4-3me IHC staining was similar between the normal controls and GC specimens.In contrast,the level of uH2B was significantly lower in the malignant gastric tissues(vs normal control tissues)and decreased along with increases in dedifferentiation(well differentiated>moderately differentiated>poorly differentiated).The level of uH2B correlated with tumor differentiation(P<0.001),Lauren’s diffuse-and intestinal-type classification(P<0.001),lymph node metastasis(P=0.049)and tumor-node-metastasis stage(P=0.005).Patients with uH2B+staining had higher 5-year survival rates than patients with uH2B-staining(52.692±2.452 vs23.739±5.207,P<0.001).The uH2B level was an independent prognostic factor for cancer-specific survival(95%CI:0.237-0.677,P=0.001).CONCLUSION:uH2B displays differential IHC staining patterns corresponding to progressive stages of GC.uH2B may contribute to tumorigenesis and could be a potential therapeutic target.

A comparative analysis of differentially expressed genes in rostral and caudal regions after spinal cord injury in rats

The initial mechanical damage of a spinal cord injury(SCI)triggers a progressive secondary injury cascade,which is a complicated process integrating multiple systems and cells.It is crucial to explore the molecular and biological process alterations that occur after SCI for therapy development.The differences between the rostral and caudal regions around an SCI lesion have received little attention.Here,we analyzed the differentially expressed genes between rostral and caudal sites after injury to determine the biological processes in these two segments after SCI.We identified a set of differentially expressed genes,including Col3a1,Col1a1,Dcn,Fn1,Kcnk3,and Nrg1,between rostral and caudal regions at different time points following SCI.Functional enrichment analysis indicated that these genes were involved in response to mechanical stimulus,blood vessel development,and brain development.We then chose Col3a1,Col1a1,Dcn,Fn1,Kcnk3,and Nrg1 for quantitative real-time PCR and Fn1 for immunostaining validation.Our results indicate alterations in different biological events enriched in the rostral and caudal lesion areas,providing new insights into the pathology of SCI.

主要胆汁的肝硬化和世袭出血性的毛细管扩张: 当二稀罕疾病共存时

Primary biliary cirrhosis is a slowly progressive cholestatic autoimmune liver disease that mainly affects middle-aged women with an estimated prevalence ranging from 6.7 to 402 cases per million. Hereditary hemorrhagic telangiectasia, or Rendu-Osler-Weber disease, is an autosomal dominant disorder characterized by angiodysplastic lesions (telangiectases and arteriovenous malformations) that can affect many organs, including liver, with a prevalence of 1-2 cases per 10000. We describe the coexistence, for the first time to our knowledge, of these two rare diseases in a 50-year old Caucasian woman. In this setting, the relevance of an accurate medical history, the role of liver histology and the characterization of liver involvement through dynamic imaging techniques can be emphasized.

Disseminated carcinomatosis of the bone marrow caused by granulocyte colony-stimulating factor:A case report and review of literature

BACKGROUND Disseminated carcinomatosis of the bone marrow(DCBM)is a widespread metastasis with a hematologic disorder that is mainly caused by gastric cancer.Although it commonly occurs as a manifestation of recurrence long after curative treatment,the precise mechanism of relapse from dormant status remains unclear.Granulocyte colony-stimulating factor(G-CSF)can promote cancer progression and invasion in various cancers.However,the potential of G-CSF to trigger recurrence from a cured malignancy has not been reported.CASE SUMMARY A 55-year-old Japanese woman was diagnosed with Ewing sarcoma localized on the fifth lumbar vertebrae 6 years after curative gastrectomy for T1 gastric cancer.After palliative surgery to release nerve compression,pathological diagnosis of the resected specimen was followed by curative radiation and chemotherapy.During treatment,G-CSF was administered 32 times for severe neutropenia prophylaxis.Eight months after completing definitive treatment,she complained of severe back pain and was diagnosed as multiple bone metastases with DCBM from gastric cancer.Despite palliative chemotherapy,she died of disseminated intravascular coagulation 13 d after the diagnosis.Immunohistochemical examination of the autopsied bone marrow confirmed a diffuse positive staining for the G-CSF receptor(G-CSFR)in the relapsed gastric cancer cell cytoplasm,whereas the primary lesion cancer cells showed negative staining for G-CSFR.In this case,G-CSF administration may have been the key trigger for the disseminated relapse of a dormant gastric cancer.CONCLUSIONWhen administering G-CSF to cancer survivors,recurrence of a preceding cancer should be monitored even after curative treatment.

Expression of the transcription factor Xvent-2 in <i>Xenopus laevis</i>embryogenesis

Till now the transcription factor Xvent-2 has been studied in Xenopus embryos only by the mRNA testing. We use immunochemical methods for testing of the Xvent-2 protein and gradient-centrifugation methods for estimation of activity of its mRNA. Our results show that the Xvent-2 protein is present in eggs and early embryos. The Xvent-2 mRNA is absent at any of these developmental stages. The majority of mRNA synthesized on the zygotic genome was stored in informosomes, while only its small part could be revealed in polysomes. The spatial patterning of the Xvent-2 protein at different developmental stages did not entirely agree with that of its mRNA. These data indicate that the Xvent-2 protein functioning in Xenopu embryos is regulated not only at the transcription, but at translation and posttranslation as well. We propose that the activation of translation on the masked Xvent-2 mRNA may lead to blood differentiation and cell migration.

Isolation of L. interrogans Serovar Pomona in 14 Human Cases and an African Lion, All with Chronic Leptospirosis

Depending on its time of evolution, leptospirosis presents two phases: acute and chronic. The chronic leptospirosis is widely accepted in animals and, until recently, this was denied in humans. Since the 1990s, we have studied dozens of cases demonstrating the chronicity of leptospirosis, including an asymptomatic or undetermined form that may remain so throughout life time or may evolve into a definitive chronic disease and/or worsen, becoming severe cases. A fundamental part for its demonstration is the isolation and characterization of the etiologic agent. In this work, the first 14 isolations of Leptospira sp. are reported from human chronic leptospirosis and an African lion diagnosed with chronic leptospirosis, obtained between 2002 and 2006, in urine, blood, and skin microbiopsies. From the 14 isolations from patients with chronic leptospirosis, one of the cases is apparently congenital and two are from patients with chronic asymptomatic leptospirosis. The diagnosis was performed by means of videorecording in darkfield microscopy in blood and urine, specific culture media, and MAT serology. Argentic impregnation and detection of specific antigen by immunostaining in blood were performed in seven patients;in two patients, skin microbiopsies and biopsies were performed. All cases were positive by video-recording in darkfield, one of them MAT was 1:160, and the rest were positive at low titers (≤1:80). The two skin biopsies were positive by argentic impregnation and immunostaining. Eleven isolates were obtained from blood, two from skin, and one from urine. The isolates were characterized by monoclonal antibodies as Leptospira serovar Pomona. L. Pomona is considered of low virulence and high adaptability to the host, and is frequently associated to chronic leptospirosis in animals. In our study, it was the only isolated serovar, including the one isolated from the lion, which would seem the most common one in our country and responsible for a large portion of cases of chronic human leptospirosis.

Clinicopathologic diagnosis of dVIN related vulvar squamous cell carcinoma:An extended appraisal from a tertiary women's hospital

Background:Differentiated vulvar intraepithelial neoplasia(dVIN)is a non-human papilloma virus(HPV)-related high-grade precursor lesion to vulvar squamous cell carcinoma(vSCCa).Although TP53 gene mutations have been identified in 80%of dVIN,its role in dVIN pathogenesis as well as malignant transformation is still being poorly understood.Poor reproducible diagnostic criteria and ambiguous p53 immunostaining patterns,along with morphologic discordance still pose a diagnostic challenge.Methods:A series of 60 cases of dVIN-related vSCCa along with adjacent dVIN were evaluated.Clinicopathological features as well as immunohistochemical results were recorded on the resection-confirmed dVIN-related vSCCa.Results:The average age of the patients was 71 years.Thirty-five cases(58.4%)of dVIN-related vSCCa were moderately differentiated,fourteen cases(23.3%)were poorly differentiated,and the remaining eleven cases(18.3%)were well-differentiated.Twenty-nine cases(48.3%)were found to have lichen sclerosus adjacent to dVIN.In terms of p53 and p16 expression in dVIN-related vSCCa and the adjacent dVIN,fifty-five(91.7%)dVIN showed mutant p53 immunostaining pattern with strong positive expression in 80%cases(basal/para-basal expression)and null pattern expression in 11.7%cases.Five(8.3%)dVIN showed p53 wild-type staining pattern.The wild-type pattern were seen in 5%of vSCCa and p53 null pattern were seen in 13.3%vSCCa.Six cases demonstrated atypical staining patterns:two cases showed p53 null expression in dVIN but p53 overexpression in invasive carcinoma;three cases exhibited p53 null expression in invasive carcinoma,with the adjacent dVIN showing basal and para-basal mutant(2 cases)and wild-type(1 case)p53 expression patterns.A single case demonstrated p53 wild-type pattern in dVIN and overexpression in invasive carcinoma.In addition,65%dVIN were p16 negative and 31.7%dVIN had patchy p16 staining.Conclusion:Clinical and prognostic value of the ambiguous/inconsistent patterns are uncertain and molecular studies are needed for further characterization.

Primary neuronal-astrocytic co-culture platform for neurotoxicity assessment of di-(2-ethylhexyl) phthalate

Plastics such as polyvinyl chlorides （PVC） are widely used in many indoor constructed environments; however, their unbound chemicals, such as di-（2-ethylhexyl） phthalates （DEHP）, can leach into the surrounding environment. This study focused on DEHP＇s effect on the central nervous system by determining the precise DEHP content in mice brain tissue after exposure to the chemical, to evaluate the specific exposure range. Primary neuronal-astrocyte co-culture systems were used as in vitro models for chemical hazard identification of DEHP. Oxidative stress was hypothesized as a probable mechanism involved, and therefore the total reactive oxygen species （ROS） concentration was determined as a biomarker of oxidative stress. In addition, NeuriteTracer, a neurite tracing plugin with ImageJ, was used to develop an assay for neurotoxicity to provide quantitative measurements of neurological parameters, such as neuronal number, neuron count and neurite length, all of which could indicate neurotoxic effects. The results showed that with 1 nmol/L DEHP exposure, there was a significant increase in ROS concentrations, indicating that the neuronal-astrocyte cultures were injured due to exposure to DEHP. In response, astrocyte proliferation （gliosis） was initiated, serving as a mechanism to maintain a homeostatic environment for neurons and protect neurons from toxic chemicals. There is a need to assess the cumulative effects of DEHP in animals to evaluate the possible uotake and effects on the human neuronal system from exoosure to DEHP in the indoor environment.

Qualitative detection of ginsenosides in brain tissues after oral administration of high-purity ginseng total saponins by using polyclonal antibody against ginsenosides

Given the limited studies and conflicting findings, the transport character of ginsenosides crossing the blood–brain barrier(BBB) remains unclear. The present study was designed to qualitatively determine the distribution of ginsenosides in brain tissues after oral administration of ginseng total saponins, using high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) combined with immunohistochemistry. In brain tissue homogenates, ginsenoside Rg1 was detectable and no other ginsenosides or their metabolites were found. No ginsenosides were detected in cerebrospinal fluid. Immunohistochemistry staining of brain tissue sections by using anti-ginsenoside polyclonal antibodies revealed the localization of ginsenosides in brain tissues. Furthermore, immunofluorescence double staining revealed that ginsenosides widely existed in vascular endotheliocytes and astrocytes, and in few neurons. These results indicated that Rg1 was the main component that entered the brain after oral administration of ginseng total saponins and that ginsenosides could cross the BBB, although the transport capability of ginsenosides through the BBB may be poor.

Laminar Distribution of Neurochemically-Identified Interneurons and Cellular Co-expression of Molecular Markers in Epileptic Human Cortex

Inhibitory GABAergic interneurons are fundamental elements of cortical circuits and play critical roles in shaping network activity. Dysfunction of interneurons can lead to various brain disorders, including epilepsy,schizophrenia, and anxiety. Based on the electrophysiological properties, cell morphology, and molecular identity,interneurons could be classified into various subgroups. In this study, we investigated the density and laminar distribution of different interneuron types and the coexpression of molecular markers in epileptic human cortex.We found that parvalbumin(PV) and somatostatin(SST)neurons were distributed in all cortical layers except layer I, while tyrosine hydroxylase(TH) and neuropeptide Y(NPY) were abundant in the deep layers and white matter.Cholecystokinin(CCK) neurons showed a high density in layers IV and VI. Neurons with these markers constituted*7.2%(PV), 2.6%(SST), 0.5%(TH), 0.5%(NPY), and4.4%(CCK) of the gray-matter neuron population. Doubleand triple-labeling revealed that NPY neurons were also SST-immunoreactive(97.7%), and TH neurons were more likely to express SST(34.2%) than PV(14.6%). A subpopulation of CCK neurons(28.0%) also expressed PV, but none contained SST. Together, these results revealed the density and distribution patterns of different interneuron populations and the overlap between molecular markers in epileptic human cortex.

Postnatal germ cell development in cryptorchid boys

Cryptorchidism is associated with infertility in adulthood.Early orchiopexy is suggested to reduce the risk.Information is lacking on the potential link between infant germ cell maturation and the risk of future infertility.The objective of the study was to evaluate age-related germ cell development in cryptorchidism.Immunostaining for markers of germ cell development(octamerbinding transcription factor 3/4[0CT3/4],placental alkaline phosphatase[PLAP],KIT proto-oncogene[C-KIT],podoplanin[D2-40],Lin-28 homolog A[LIN28],and G antigen 7[GAGE-7])was performed in testicular biopsies from 40 cryptorchid boys aged 4-35 mon ths.Germ cell nu mbers and distributi ons were evaluated in cross sectio ns of semi niferous tubules,with and without immuno staini ng.OCT3/4,D2-40,and LIN 28 were gen erally expressed in the early stages of germ cell development,as show n by positive expression in germ cells in the central region of seminiferous tubules・In contrast,PLAP and GAGE-7 were expressed in both central and peripheral parts of the tubules in the early stages of development and expressed mainly in a peripheral position with advancing age.Germ cell maturation was delayed in this study population as compared with that observed in our previous study on germ cell markers in a healthy population.The number of GAGE-7-positive germ cells per tubular cross section obtained by immunostaining was significantly higher than that obtained by standard hematoxylin and eosin staining.Double immunostaining revealed heterogeneity in germ cell development in cryptorchid testes.These results shed light on the pathophysiology of germ cell development in boys with cryptorchidism.

Expression of programmed death-1 and its ligands in the liver of biliary atresia

Background:An aberrant immune response is the predominant pathogenetic factor in biliary atresia (BA).Programmed death-1 (PD-1) and its two ligands,programmed death ligand-1 and programmed death ligand-2 (PD-L1 and PD-L2,respectively) play an important inhibitory role in immune reactions.We aimed to illustrate the expression of these molecules in BA.Methods:Liver specimens were obtained from infants with BA during the Kasai procedure (early BA) and fiver transplantation (late BA).Intrahepatic expression of PD-1,PD-L1,and PD-L2 were examined by immunostaining and compared with that in patients with neonatal hepatitis syndrome and normal controls.The correlation between the expression levels of these molecules in the liver and clinicopathological parameters was analyzed for each group.Results:Enhanced expression of PD-1 and its ligands occurred in the livers with early BA.In the BA-affected livers,PD-1 was correlated with the degree of peri-bifiary inflammation,while PD-L2 was linked more directly with portal fibrosis.None of the three molecules was correlated with the prognosis of the Kasai procedure in patients with early BA.Conclusion:Only PD-1 and PD-L1 are involved in the immune reactions of early BA.Elucidation of the detailed role of PD-L2 in BA requires further research.

Florid cystic endosalpingiosis of the uterine subserosa: A case report

Dear Editor:Florid cystic endosalpingiosis(FCE)is a rare type of endosalpingiosis that presents as a mass-like lesion.It may rarely involve the uterus and present as a cystic or tumor-like mass.Herein we report an unusual case of FCE.The patient is a 43-year-old woman presented with pelvic cystic mass,which was diagnosed as an ovarian cyst by ultrasonic examination,and diagnosed as a uterine leiomyoma with cystic degeneration by laparoscopy.Pathologic findings show FCE of the uterine subserosa.Preoperative diagnosis of uterine cystic endosalpingiosis is usually difficult.Thus,awareness about this condition in clinicians may help in preventing misdiagnosis and overtreatment.A 43-year-old woman,gravida 6 para 1,presented a cystic formation at the left adnexal area.