Combination of Tertiary Lymphoid Structure and Neutrophil-to-Lymphocyte Ratio Predicts Survival in Patients With Hepatocellular Carcinoma

BackgroundHepatocellular carcinoma (HCC) is the most common pathological type of primary liver cancer. The lack of prognosis indicators is one of the challenges in HCC. In this study, we investigated the combination of tertiary lymphoid structure (TLS) and several systemic inflammation parameters as a prognosis indicator for HCC.Materials and MethodsWe retrospectively recruited 126 postoperative patients with primary HCC. The paraffin section was collected for TLS density assessment. In addition, we collected the systemic inflammation parameters from peripheral blood samples. We evaluated the prognostic values of those parameters on overall survival (OS) using Kaplan-Meier curves, univariate and multivariate Cox regression. Last, we plotted a nomogram to predict the survival of HCC patients.ResultsWe first found TLS density was positively correlated with HCC patients’ survival (HR=0.16, 95% CI: 0.06 − 0.39, p < 0.0001), but the power of TLS density for survival prediction was found to be limited (AUC=0.776, 95% CI:0.772 − 0.806). Thus, we further introduced several systemic inflammation parameters for survival analysis, we found neutrophil-to-lymphocyte ratio (NLR) was positively associated with OS in univariate Cox regression analysis. However, the combination of TLS density and NLR better predicts patient’s survival (AUC=0.800, 95% CI: 0.698-0.902, p < 0.001) compared with using any single indicator alone. Last, we incorporated TLS density, NLR, and other parameters into the nomogram to provide a reproducible approach for survival prediction in HCC clinical practice.ConclusionThe combination of TLS density and NLR was shown to be a good predictor of HCC patient survival. It also provides a novel direction for the evaluation of immunotherapies in HCC.

Nested PCR for the Diagnosis of Feline Sporotrichosis From Formalin-Fixed and Paraffin-Embedded Samples Using Different DNA Extraction Protocols

Sporotrichosis is a chronic, cosmopolitan granulomatous mycosis that affects humans and animals. The infection is caused by the dimorphic fungi Sporothrix sp. The aims of the present study were to evaluate, standardize and validate a nested PCR technique using two DNA purification kits for the extraction of DNA from formalin fixed and paraffin-embedded tissues (FFPE) for Sporothrix sp. detection. FFPE mycological culture pellet samples of different Sporothrix species (S. chilensis, S. mexicana, S. pallida, S. globosa, S. brasiliensis and S. schenckii) were used as positive controls and clinical FFPE tissue samples of animals positive for Cryptococcus sp., Leishmania infantum and Histoplasma sp. were used as negative controls. Ten clinical FFPE skin samples from cats with sporotrichosis were used to validate the nested PCR. These samples were cut into two distinct paraffin sectioning protocols (5 and 16 μm thick). The paraffin sections were subjected to two different DNA extraction kits (chemical and thermal extractions). A nested PCR was performed on the extracted DNA to identify the genus Sporothrix. The chemical extraction protocol with the 5 μm thick paraffin section was more effective in extracting DNA from Sporothrix sp. from FFPE samples and the nested PCR technique showed the highest sensitivities (100% in the positive controls and of 50% in the skin samples of cats) and specificity (100%). Therefore, the nested PCR using this protocol has great potential to be applied in Sporothrix sp. diagnosis in FFPE samples of cats.

Borderline ovarian tumor frozen section diagnoses with features suspicious of invasive cancer: a retrospective study

Background A frozen section diagnosis of a borderline ovarian tumor with suspicious features of invasive carcinoma (“at least borderline” or synonymous descriptions) presents us with the dilemma of whether or not to perform a full ovarian cancer staging procedure. Quantification of this dilemma may help us with the issue of this clinical decision. The present study assessed and compared both the prevalence of straightforward borderline and “at least borderline” frozen section diagnoses and the proportion of these women with a final histopathological diagnosis of invasive carcinoma, with a special interest in histologic subtypes. Methods A retrospective cohort study was performed in three hospitals in The Netherlands. All women that underwent ovarian surgery with perioperative frozen section evaluation in one of these hospitals between January 2007 and July 2018 were identified and included in case of a borderline or “at least borderline” frozen section diagnosis and a borderline ovarian tumor or invasive carcinoma as a final diagnosis. Results A total of 223 women were included, of which 41 women (18.4%) were diagnosed with “at least borderline” at frozen section. Thirteen of forty-one women (31.7%) following “at least borderline” frozen section diagnosis and 14 of 182 women (7.7%) following a straightforward borderline frozen section diagnosis were diagnosed with invasive carcinoma at paraffin section evaluation (p < 0.001). When compared to straightforward borderline frozen section diagnoses, the proportion of women diagnosed with invasive carcinoma increased from 3.1 to 35.7% for serous tumors (p = 0.001), 10.0 to 21.7% for mucinous tumors (p = 0.129) and 50.0 to 75.0% (p = 0.452) in case of other histologic subtypes following an “at least borderline” frozen section diagnosis. Conclusions Overall, when compared to women with a decisive borderline frozen section diagnosis, women diagnosed with “at least borderline” frozen section diagnoses were found to have a higher chance of carcinoma upon final diagnosis (7.7% vs 31.7%). Especially in the serous subtype, full staging during initial surgery might be considered after preoperative consent to prevent a second surgical procedure or chemotherapy in unstaged women. Further studies are needed to evaluate whether additional sampling in case of an “at least borderline” diagnosis may decrease the risk of surgical over-treatment.

Pharmacognostic Identification of the Green Peel of Juglans mandshurica Maxim. from Different Regions and Harvest Times

Background: The green peel of Juglans mandshurica Maxim. (QLY) can be used as medicine, also known as Qinglongyi[1]. The aim of this study was to identify QLY from the different regions and harvest times by pharmacognostic identification.Methods: In this study, the morphological character, microscopic character and juglone’s content of QLY from different regions and harvest times were compared by morphological identification, paraffin section and powder section, physico-chemical identification and High Performance Liquid Chromatography (HPLC).Results: Morphological identification results that the colour of outer surfaces of peels which harvested from Tangyuan county of Heilongjiang Province in July, August and September are respectively yellow brown, tan and black brown , and the color of the solution graded gradually from light brown yellow to brownish black by water-based test (WBT). The colour of outer surfaces of peels which harvested from Changbai county of Jilin Province in July, August and September are respectively yellowish white, brownish yellow and pale brown, and the color of the solution gradually changed from light brown to dark brown by WBT. As well as the colour of outer surfaces of peels which harvested from Qingyuan county of Liaoning Province in July, August and respectively September are Light yellowish green, light brownish green and brown green, and the color of the solution gradually changed from light green to dark brown by WBT. With the increase of growth period, the number of microscopic characteristics of different Qinglongyi increased. Through physico-chemical identification, it was found that the yield of juglone in QLY picked on September 21, 2016 was the highest, and the content of juglone, whose order was Heilongjiang > Jilin > Liaoning, was as high as 203.476 μg/g.Conclusions: Through comparison, it is concluded that Heilongjiang Province is one of the high-quality producing areas of QLY, and its best harvest time is in the middle and first ten-day period of September to harvest QLY without softening, yellowing and decay. This experiment provides a theoretical basis for the determination of the best harvest conditions in QLY and the establishment of identification standards for medicinal materials.

A case report: pseudoxanthoma elasticum diagnosed based on ocular angioid streaks and the curative effect of Conbercept treatment

Background This article is a case report of pseudoxanthoma elasticum (PXE) which was diagnosed based on significant angioid streaks (AS) with choroidal neovascularization (CNV) and regain normal visual function by intravitreal injection with Conbercept. Case presentation A 51-year-old woman was referred to the Ophthalmology Department of Qingdao Municipal Hospital (Qingdao, China) on September 14, 2020 for metamorphopsia and loss of vision in the left eye in the preceding three days. Past history: high myopia for more than 30 years, best corrected visual acuity (BCVA) of both eyes was 1.0 (5 m Standard Logarithm Visual Acuity chart in decimal notations), hypertension for six years, and cerebral infarction two years ago, no history of ocular trauma or surgeries or similar patients in family was documented. We used methods for observation, including fundus examination, optical coherence tomography (OCT), fluorescein angiography combined with indocyanine green angiography (FFA + ICGA). Due to her symptoms and manifestations, along with the appearance of her neck skin, which resembled ‘chicken skin’, we speculated that she should be further examined at the Department of Dermatology by tissue paraffin section and molecular pathology analyses, and the diagnosis of PXE was then confirmed. After intravitreal injection with Conbercept (10 mg/ml, 0.2 ml, Chengdu Kanghong Biotechnologies Co., Ltd.; Chengdu, Sichuan, China) she regained her BCVA. Conclusions This patient regained her best corrected visual acuity through intravitreal injection with Conbercept. To the best of our knowledge, no publications are available on cases in which a vision loss and the normal visual function can be reverted by intravitreal injection with Conbercept. Although PXE is a disease with low incidence and thus no effective cure established, targeted symptomatic treatment can effectively retard the disease progression and improve visual function, such as intravitreal injection with Conbercept.

Morphological changes and functional circRNAs screening of rabbit skeletal muscle development

Background The temporal expression pattern of circular RNAs (circRNAs) across developmental stages is essential for skeletal muscle growth and functional analysis. However, there are few analyses on the potential functions of circRNAs in rabbit skeletal muscle development. Results Initially, the paraffin sections showed extremely significant differences in the diameter, number, area and density of skeletal muscle fibers of the fetus, child, adult rabbit hind legs (P < 0.01). Then, RNA-seq libraries of these three stages were constructed. A total of 481 differentially expressed circRNAs (DE-circRNAs) and 5,658 differentially expressed genes (DEGs) were identified. Subsequently, DE-circRNAs, whose host genes were DEGs or non-DEGs, were analyzed by GO respectively. In the fetus vs. child group, up-regulated DE-circRNAs (whose host genes were DEGs) were related to muscle fiber structure, and down-regulated ones were related to mitosis. The up-regulated DE-circRNAs (whose host genes were non-DEGs) were involved in enzyme activity, methylation and glycosylation, and the down-regulated ones were involved in mitosis and catabolism. In the fetus vs. adult group, the up-regulated DE-circRNAs (whose host genes were DEGs) were related to skeletal muscle basic structure, and the down-regulated ones were also associated with cell proliferation. But the up-regulated DE-circRNAs (whose host genes were non-DEGs) were connected with regulation of histone ubiquitination, chromatin and organelles. The down-regulated DE-circRNAs were connected with the catabolism processes. In addition, novel_circ_0022663 and novel_circ_0005489, which might have coding potential, and novel_circ_0004210 and novel_circ_0001669, which might have miRNA sponge capability, were screened out. Conclusions In this study, hind leg muscles of fetus, child and adult rabbits were collected for paraffin section and RNA-seq to observe the structural changes of skeletal muscle and obtain circRNA expression profiles at different stages. These data provided a catalog of circRNAs related to muscle development in New Zealand rabbits, allowing us to better understand the functional transitions in mammalian muscle development.

Sentinel Lymph Node Biopsy Using a Single-Dye Technique in a Cancer Center of North-East India

Background Sentinel lymph node (SLN) is the first node to receive the drainage directly from a tumor. SLN biopsy can be done in lieu of a formal lymphadenectomy in selected clinically node-negative cancers and minimizes morbidity compared with the latter. Methods This prospective study was done in patients with operable clinically node-negative breast cancer, penile cancer, and malignant melanoma of extremities in a cancer center of North-east India from January 2019 to December 2019. All the patients underwent formal lymph nodal dissection after the SLN biopsy. Besides intraoperative frozen section study of the sentinel node(s), all the specimens, including the sentinel node(s), were subjected to paraffin section histopathology. Results SLN was identified successfully in 96% of patients. Mean number of sentinel node(s) dissected was 2.3. Study of SLN biopsy with methylene blue dye for staging was done with 100% sensitivity and 95.3% specificity. The SLN procedure was able to negatively predict the drainage nodal basin in 100% with an overall accuracy of staging of 96.5%. The true-positive rate noted was 88.8%, and the false-positive rate was 4.6%. Conclusions SLN using a single-dye technique reliably identifies a sentinel node. This procedure can be safely adopted in patients with node-negative cancers as mentioned above to pathologically study the drainage basin.

Mechanisms of thermal treatment on two dominant copepod species in O3/BAC processing of drinking water

Phyllodiaptomus tunguidus and Heliodiaptomus falxus are dominant copepods species in drinking water processing plants in southern China. With a potential penetration risk, the breeding and leakage of copepods are drawing more and more attention in recent years. The current study provided a thermal treatment method to control copepods and their eggs. Results showed that: (1) the immediate death rates of P. tunguidus and H. falxus after heated to 34–40 °C for 5 min are positively correlated to the treatment temperatures (P < 0.01), and all individuals of the both species were eliminated after heated at 40 °C for 5 min; (2) overall hatching rates of P. tunguidus eggs were negatively correlated with treatment temperatures (P < 0.01) between 39–45 °C, with zero percent hatched after treatment at 45 °C for 5 min; (3) hatching rates of H. falxus were negatively correlated with treatment temperatures (P < 0.01) between 37–41 °C, with no nauplii hatched when treated at 41 °C for 5 min; (4) paraffin section histological examination indicated that thermal treatment caused severe damage to internal organs and egg structure. Finally, based on the experimental data, the application of the thermal treatment method was discussed in ozonation combined with biological activated carbon (O3/BAC) processing of drink water treatment.

To Determine the Accuracy of Frozen Section in the Management of Ovarian Tumours

BACKGROUND Frozen section done during surgery provides a valuable tool in the diagnosis and classification of tumours into benign, borderline and malignant. In this study, we evaluated the intraoperative findings and the accuracy of the frozen section and compared it with the final histopathology report. METHODS A retrospective study was done in the Department of Obstetrics and Gynecology in Sri Ramachandra University and Research Centre from January 2017 to January 2020, for a period of 3 years and data was taken from the patients who underwent surgery-staging laparotomy for ovarian masses along with frozen section in the diagnosis of their tumor. The final histopathology report is the gold standard for detection of ovarian tumours. The accuracy of the frozen section was compared with the final histopathological report on paraffin section. RESULTS There was a total of 109 cases during the study period of 3 years. Out of the total, frozen section showed 55 malignancy cases, 7 cases were borderline, and 47 cases were benign tumors; whereas the final histopathology report showed 51 malignant cases, 9 borderline cases, and 49 cases were benign tumors. This shows that frozen section has almost 97 % accuracy. CONCLUSIONS Frozen section is a very good tool in the diagnosis of tumour whether benign, borderline or malignant and when compared with the final histopathological report gives good accuracy. The diagnosis given during surgery is helpful in the further management of patients according to the stage of the tumour. KEY WORDS Frozen Section, Benign, Borderline, Malignant tumours, Final Histopathological Report