Filtered Saliva for Rapid and Accurate Analyte Detection for POC Diagnostics.

Saliva has shown considerable promise as a diagnostic medium for point-of-care (POC) and over-the-counter (OTC) diagnostic devices due to the non-invasive nature of its collection. However, a significant limitation of saliva-based detection is undesirable interference in a sensor's readout caused by interfering components in saliva. In this study, we develop standardized sample treatment procedures to eliminate bubbles and interfering molecules while preserving the sample's target molecules such as spike (S) protein and glucose. We then test the compatibility of the pretreatment system with our previously designed SARS-CoV-2 and glucose diagnostic biosensing systems for detecting S protein and glucose in subject saliva. Ultimately, the effectiveness of each filter in enhancing biomarker sensitivity is assessed. The results show that a 20 mg nylon wool (NW) filter shows an 80% change in viscosity reduction with only a 6% reduction in protein content, making it an appropriate filter for the salivary S protein diagnostic system. Meanwhile, a 30 mg cotton wool (CW) filter is identified as the optimal choice for salivary glucose detection, achieving a 90% change in viscosity reduction and a 60.7% reduction in protein content with a minimal 4.3% reduction in glucose content. The NW pretreatment filtration significantly improves the limit of detection (LOD) for salivary S protein detection by five times (from 0.5 nM to 0.1 nM) and it reduces the relative standard deviation (RSD) two times compared to unfiltered saliva. Conversely, the CW filter used for salivary glucose detection demonstrated improved linearity with an R2 of 0.99 and a sensitivity of 36.6 µA/mM·cm2, over twice as high as unfiltered saliva. This unique filtration process can be extended to any POC diagnostic system and optimized for any biomarker detection, making electrochemical POC diagnostics more viable in the current market.

Metaanálisis sobre el protocolo de recuperación acelerada tras la cistectomía radical / Enhanced pharmaceutical recovery as postoperative standard care after radical cystectomy: A meta-analysis

Introducción: Se realizó un metaanálisis para evaluar el efecto de los protocolos de recuperación acelerada después de la cistectomía radical.Métodos: Se realizó una búsqueda bibliográfica sistemática hasta abril de 2021 y se incluyeron 33 estudios con 6.596 sujetos tratados mediante cistectomía radical; 3.143 de ellos fueron sometidos a protocolos de recuperación acelerada tras la cirugía y 3.453 eran controles. Los estudios incluían informes sobre los efectos del protocolo de recuperación acelerada tras la cirugía de cistectomía radical. Se calculó el odds ratio (OR) y la diferencia de medias (DM) con intervalos de confianza (IC) del 95% para evaluar los efectos del protocolo de recuperación acelerada tras la cistectomía radical mediante los métodos dicotómicos y continuos modelos de efectos aleatorios o fijos.Resultados: En los sujetos que recibieron el protocolo de recuperación acelerada después de la cistectomía radical se redujeron significativamente la duración de estancia hospitalaria (DM: −2,78; IC del 95%: −3,59 a −1,97, p<0,001), la tasa de complicaciones (OR: 0,75; IC del 95%: 0,60 a 0,94, p=0,01), el reingreso en un plazo de 30días (OR: 0,80; IC del 95%: 0,69 a 0,94, p=0,007) y el tiempo hasta la recuperación de la función intestinal (DM: −1,30; IC del 95%: −2,22 a −0,37, p=0,006), en comparación con los controles.Conclusiones: La aplicación de protocolos de recuperación acelerada después de la cirugía puede asociarse a una reducción de la estancia hospitalaria, complicaciones, reingreso en 30días y tiempo hasta la recuperación de la función intestinal tras la cistectomía radical. Sin embargo, se requieren más estudios para validar estos hallazgos. (AU)

Introduction: We performed a meta-analysis to evaluate the effect of enhanced pharmaceutical recovery as postoperative standard care after radical cystectomy.Methods: A systematic literature search up to April 2021 was done and 33 studies included 6596 subjects submitted to surgery for radical cystectomy at the start of the study; 3143 of them received enhanced pharmaceutical recovery after surgery and 3453 were controls. The studies reported relationships about the effects of enhanced pharmaceutical recovery as postoperative standard care after radical cystectomy. We calculated the odds ratio (OR) and mean difference (MD) with 95% confidence intervals (CIs) to assess the effects of enhanced pharmaceutical recovery as postoperative standard care after radical cystectomy using the dichotomous and continuous methods with a random or fixed-effect model.Results: Enhanced pharmaceutical recovery after surgery had significantly lower length of hospital stay (MD: −2.78; 95%CI: −3.59 to −1.97, P<.001), complications (OR: 0.75; 95%CI: 0.60 to 0.94, P=.01), readmission within 30days (OR: 0.80; 95%CI: 0.69 to 0.94, P=.007), and time to defecation (MD: −1.30; 95%CI: −2.22 to −0.37, P=.006) compared to control in subjects submitted to radical cystectomy.Conclusions: Enhanced pharmaceutical recovery after surgery may reduce the length of hospital stay, complications, readmission within 30days, and time to first bowel movement compared to control in subjects with surgery for radical cystectomy. Furthers studies are required to validate these findings. (AU)

Deleterious effects of SARS-CoV-2 infection on human pancreatic cells

COVID-19 pandemic has infected more than 46 million people worldwide and caused more than 1.2 million deaths. It is transmitted by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and affects the respiratory tract as well as extra-pulmonary systems, including the pancreas, that express the virus entry receptor, Angiotensin-Converting Enzyme 2 (ACE2) receptor. Importantly, the endocrine and exocrine pancreas, the latter composed of ductal and acinar cells, express high levels of ACE2, which correlates to impaired functionality characterized as acute pancreatitis observed in some cases presenting with COVID-19. Since acute pancreatitis is already one of the most frequent gastrointestinal causes of hospitalization in the U.S. and the majority of studies investigating the effects of SARS-CoV-2 on the pancreas are clinical and observational, we utilized human iPSC technology to investigate the potential deleterious effects of SARS-CoV-2 infection on iPSC-derived pancreatic cultures containing endocrine and exocrine cells. Interestingly, SARS-CoV-2 is capable of infecting iPSC-derived pancreatic cells, thus perturbing their normal molecular and cellular phenotypes. The infection increased a key inflammatory cytokine, CXCL12, known to be involved in pancreas dysfunction. Transcriptome analysis of infected pancreatic cultures confirmed that SARS-CoV-2 hijacks the ribosomal machinery in these cells. Notably, the SARS-CoV-2 infectivity of the pancreas is confirmed in post-mortem tissues from COVID-19 patients, which showed co-localization of SARS-CoV-2 in pancreatic endocrine and exocrine cells and increased the expression of some pancreatic ductal stress response genes. Thus, we demonstrate for the first time that SARS-CoV-2 can directly infect human iPSC-derived pancreatic cells with supporting evidence of presence of the virus in post-mortem pancreatic tissue of confirmed COVID-19 human cases. This novel model of iPSC-derived pancreatic cultures will open new avenues for the comprehension of the SARS-CoV-2 infection and potentially establish a platform for endocrine and exocrine pancreas-specific antiviral drug screening.

The phosphorylation of hCDC14A modulated by ZIPK regulates autophagy of murine pancreatic islet ß-TC3 cells upon glucose stimulation.

OBJECTIVE: Depletion of islet ß cells plays a crucial role in the onset of diabetes mellitus. Cell autophagy, as a self-healing process, contributes to maintaining metabolic homeostasis and can protect islet ß cells from apoptosis upon starvation or high glucose stress. However, the underlying regulatory network of the autophagic process in islet ß cells has not been fully explored. MATERIALS AND METHODS: Murine ß-TC3 cells treated with different concentrations of glucose, and wild-type or the Ser484 mutant human cell division cycle gene 14A (hCDC14A) was transfected. Cell viability, proliferation and autophagy as well as islet secretion were studied. The mTOR and AMPK signaling pathways were investigated by western blots. Zipper-interacting protein kinase was studied using mass spectrometry and immunoprecipitation. RESULTS: Overexpression of wild-type hCDC14A, but not the Ser484 mutant hCDC14A, promoted cell viability, proliferation and autophagy accompanied by enhanced islet secretion and reduced cell apoptosis via mTOR pathway inhibition as well AMPK pathway activation in ß-TC3 cells and vice versa. Furthermore, Zipper-interacting protein kinase (ZIPK), also known as DAPK3, was found to interact with hCDC14A primarily for Ser484 phosphorylation, and ZIPK knockdown could affect the phosphorylation of hCDC14A and weaken cell death or cell cycle modulation. CONCLUSIONS: Taken together, our results may provide new insight into the role of hCDC14A in the autophagy of islet ß cells and suggest the potential therapeutic value of hCDC14A phosphorylation in the prevention and treatment of diabetes.

Distribution characteristics of hepatitis C virus genotypes in Xinjiang / 中华肝脏病杂志

To understand the hepatitis C virus (HCV) genotype distribution characteristics in Xinjiang region. 6462 cases with chronic HCV infection that were treated at the First Affiliated Hospital of Xinjiang Medical University from January 2013 to September 2018 were selected, and repeated testers were excluded. A total of 4773 cases with HCV genotypes were efficiently included. PCR-reverse dot hybridization method was used to retrospectively analyze the genotypes distribution. (2) test or F-test was used for intergroup comparison. < 0.05 was considered as statistically significant. Five genotypes were detected in 4 773 samples: genotype 1b 2928 cases (61.3%), genotype 2a 1241 cases (26%), and genotype 3a 375 cases (7.9%), genotype 3b 205 cases (4.3%), and genotype 6a 24 cases (0.5%). Patients were 48.14 ± 13.93 years old. Genotype 1b was mainly detected in all age groups. There were 2 965 cases of Han ethnicity and 1808 cases of 19 ethnic minorities, of which 1798 cases (60.6%) and 1130 cases (62.5%) were genotype 1b, and 235 cases (7.9%) and 345 cases (19.1%) were genotype 3, respectively. Among the ethnic minorities, Uyghur were the predominant, and genotype 6a could be detected; however, no other ethnic groups had detected genotype 6a. There were 704 Uyghur of genotype 1b (62.1%), 269 Uyghur of genotype 3 (23.7%), and 235 Hans of genotype 3 (7.9%). There were 2 413 males and 2 360 females, of which 1 418 males (58.8%), and 1 510 females (64%) were of genotype 1b, and 424 males (17.6%), and 156 females (6.6%) were of genotype 3. There was a statistically significant difference between the gender of patients with genotype 1b and non-genotype 1b ( < 0.05). There was a statistically significant difference in the detection rate of genotype 2a, 3a, 3b, 6a between Han and ethnic minority patients ( < 0.05). HCV genotypes distribution in Xinjiang region is diverse, and is mainly type 1b. An ethnic minority has higher proportion of HCV genotype 3 than that of Han ethnicity. HCV genotypes detection in Xinjiang region enriches the distribution characteristics of HCV genotypes and provides a basis for individualizing treatment for patients in China.

MiRNA-485-5p suppresses the proliferation of acute myeloid leukemia via targeting SALL4.

OBJECTIVE: To examine the expression level of microRNA-485-5p (miRNA-485-5p) in acute myeloid leukemia (AML) and its biological function in regulating the proliferative ability of AML through targeting SALL4. PATIENTS AND METHODS: Serum level of miRNA-485-5p in AML patients and healthy controls was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). MiRNA-485-5p level in AML cell lines was detected by qRT-PCR as well. Proliferative and apoptotic changes in AML5 and U937 cells overexpressing miRNA-485-5p were assessed. Subsequently, the regulatory effect of miRNA-485-5p on SALL4 level was evaluated. Rescue experiments were conducted to uncover the role of miRNA-485-5p/SALL4 regulatory loop in regulating cellular behaviors of AML. RESULTS: Compared with healthy controls, serum level of miRNA-485-5p was lower in AML patients. MiRNA-485-5p was similarly downregulated in AML cell lines. Overexpression of miRNA-485-5p stimulated proliferation and alleviated apoptosis in AML. SALL4 level was downregulated by transfection of miRNA-485-5p mimics in AML5 and U937 cells. Overexpression of SALL4 could reverse the regulatory effect of miRNA-485-5p on proliferative and apoptotic abilities of AML. CONCLUSIONS: MiRNA-485-5p is downregulated in AML. Overexpression of miRNA-485-5p alleviates the malignant progression of AML through downregulating SALL4.

The clinical value of China Savin pollen extract used for skin prick test / 临床耳鼻咽喉头颈外科杂志

The aim of this study is to evaluate the effectiveness and safety of China Savin pollen extract which was used for skin prick test (SPT) in the diagnosis of China Savin pollen allergy. Patients with diagnosis of allergic diseases were collected from Allergy Department of Peking Union Medical College Hospital. All patients were given SPT with China Savin pollen extract, and the mean wheal diameter (MWD) was measured after 15 minutes. Receiver operating characteristic curve (ROC) analysis was performed based on the results of serum specific immunoglobulin E (sIgE). The effectiveness of SPT in the diagnosis of China Savin pollen allergy was evaluated under different diagnostic cutoff values. Adverse events were also recorded to evaluate the safety. A total of 1 029 patients were enrolled in this study without drop out case. There were 1 007 patients in full analysis set (FAS) and 765 patients in per protocol analysis set (PPS). The elimination rate was 25.66%. The area under the ROC curve of FAS is 0.814 (95%: 0.788-0.839); which of PPS is 0.829 (95%: 0.801-0.857). Based on the ROC curve of PPS, the optimal and the 95% specificity diagnostic cutoff values of MWD were 3.25 mm and 4.75 mm respectively. Based on different diagnostic cutoff value (3.00, 3.25 and 4.75 mm), the sensitivities of SPT with China Savin pollen extract were 0.740 0 (95%: 0.701 6-0.778 4), 0.700 (95%: 0.659 8-0.740 2) and 0.532 (95%: 0.488 3-0.575 7) respectively, whereas the specificity was gradually increased in sequence, which was 0.769 8 (95%: 0.719 1-0.820 5), 0.826 4 (95%: 0.780 8-0.872 0) and 0.950 9 (95%: 0.924 9-0.976 9) respectively. There were 7 adverse events observed among 6 patients (rate: 0.583%, 6/1 029). The manifestation was mild. There was no severe adverse event. SPT with China Savin pollen extract is an effective and safe tool for the diagnosis of China Savin pollen allergy. The effectiveness of diagnosis could be improved based on integration of medical history and different diagnostic threshold values of SPT.

Risk factors of liver metastasis from advanced pancreatic adenocarcinoma: a large multicenter cohort study.

BACKGROUND: Clinical prognostic parameters of liver metastasis from pancreatic adenocarcinoma have not been specifically identified.This study is to explore the risk factors of liver metastasis in advanced pancreatic adenocarcinoma (PDAC) patients in China. METHODS: A multicenter cohort study was conducted to explore whether liver metastasis in locally advanced and metastatic PDAC could be reflected by some common laboratory indexes. We collected 1787 advanced PDAC patients from three participating hospitals between 2004 and 2014. The associations between some laboratory indexes and risks of liver metastases were analyzed. RESULTS: Results have shown that 87% of stage IV patients developed synchronous liver metastasis. Primary tumor location (body/tail vs. head/neck, OR 0.55, 95% CI 0.36-0.83), primary tumor diameter (≥20 mm vs. <20 mm, OR 1.77, 95% CI 1.16-2.70), elevated ALT and AST (OR 1.62, 95% CI 0.92-2.83), and elevated CA19-9 (OR 2.72, 95% CI 1.85-3.99) upon diagnosis are significantly associated with risk of synchronous liver metastasis. Among stage III patients, 30.1% developed metachronous liver metastasis. However, no risk factors were identified among these patients. CONCLUSIONS: Primary tumor location, diameter, elevated ALT and AST, and increased CA19-9 are independent risk factors of synchronous liver metastasis in PDAC patients.

Development and application of Real-time PCR assay for detection of Theilera annulata infection in cattle in Xinjiang Uygur Autonomous Region, China

Theileria annulata (T. annulata), the causative agent of tropical theileriosis, is a protozoan parasite that also causes lymphoproliferative diseases in cattle. Development of reliable and fast methods are necessary in the epidemiological investigation of T. annulata in ticks and animals. Real-time PCR possesses merits of rapidity, accuracy, reliability, automation and ease of standardization, which is widely used for the detection of blood borne parasites. In this study, species-specific primers and TaqMan probe were designed on the basis of the 18s rRNA gene sequence of T. annulata, and the real-time PCR assay was developed by optimizing the reaction parameter. The performance of real-time PCR was assessed by testing 47 blood samples from cattle and comparing with the results from conventional PCR. The results show that this real-time PCR assay could specifically detect 10 copies DNA of T. annulata, which is 10-fold sensitivity more than conventional PCR. No cross-reactions were observed with Babesia bovis, Babesia bigemina, Trypanosoma evansi and Theileria equi. Of the 47 field samples collected from Xinjiang Uygur Autonomous Region, China, 36.17% were detected by real-time PCR, and 25.53% were found positive for T. annulata infection by conventional PCR. These results indicated that the real-time PCR assay is a useful approach for detecting T. annulata infections and has potential as an alternative tool for ecological and epidemiological surveillance of ovine theileriosis.

Analysis of Structures, Functions, and Epitopes of Aminopeptidase from Trichinella spiralis

We have previously reported that the recombinant T. spiralis aminopeptidase (rTsAP) could induce a partial protective immunity against T. spiralis infection in mice. The aim of this study was to predict the structures and functions of TsAP protein by using the full length cDNA sequence of TsAP gene. TsAP sequence was 1515 bp length with a 1515 bp biggest ORF encoding 504-amino acid protein. The molecular weight and isoelectric point of TsAP were 54.7 kDa and 6.69, respectively. TsAP structure domains contained a Peptidase_M17_N and a Peptidase_M17 domain, which has the function of catalysis of the hydrolysis of N-terminal amino acid residues. TsAP had no signal peptide site and transmembrane domain, and located in cytoplasm. The secondary structure of TsAP contained 16 α-helix, 14 β-strand and 29 coils. The TsAP had 11 and 21 potential antigenic epitopes of T cell and B cell, respectively. Based on the phylogenetic analyses of TsAP, T. spiralis have the closest relationship with Plasmodium falciparum. TsAP was a kind of proteolytic enzyme with a variety of biological functions and its antigenic epitopes could provide important insights on the diagnostic antigens and target molecular of anti-Trichinella drugs

Sequence variation and bioinformatics analysis of Toxoplasma gondii GRA16 Gene

Toxoplasmosis is caused by the intracellular protozoan Toxoplasma gondii. It is anopportunistic zoonosis in warm-blooded animals and humans, with a worldwide distribution. Toxoplasma gondii dense granule protein 16 (TgGRA16) can modulate some functions in host cells and is considered a significant virulent factor of the parasite. The present study reports sequence variation in TgGRA16 gene among T. gondii strains from different hosts and geographical locations, and the construction of phylogenetic relationships of these T. gondii strains based on sequences of TgGRA16, and analysis of B cell epitopes in TgGRA16. Our results showed that all TgGRA16 gene sequences were 1518 bp and the C+G contents ranged from 52.17% to 52.59%. Sequence variation in the TgGRA16 gene was 0-1.51%. Phylogenetic analysis revealed that TgGRA16 gene sequence could not be used to differentiate the different T. gondii genotypes. Six B cell epitopes were predicted in TgGRA16. These results indicated that TgGRA16 gene is not an ideal marker for studying genetic relationships of T. gondii isolates, but may represent a good vaccine candidate against toxoplasmosis.

Curcumin inhibits 19-kDa lipoprotein of Mycobacterium tuberculosis induced macrophage apoptosis via regulation of the JNK pathway.

Recently, synthetic curcumin analogs are reported as potential active compounds against Mycobacterium tuberculosis (MTB). During the process of MTB infection, macrophages show increased apoptosis. The candidate virulence factors such as 19-kDa lipoprotein secreted by the MTB (P19) strongly influences macrophages by activation of Toll-like receptor 2 (TLR2) and mitogen-activated protein kinases (MAPKs). It has been reported that curcumin could affect the apoptosis of tumor cells via regulation of MAPKs. However, its effect on the P19-induced apoptosis of macrophages is unclear. This study investigates the effect of curcumin on the MAPKs signaling and apoptosis in human macrophages. The results showed that curcumin and P19 induced macrophage apoptosis in a time- and dose-dependent manner Low doses of curcumin (10 and 20 µM) protected macrophages from P19 induced apoptosis, accompanied by decreased production of cytokines and reduced activation of the c-Jun amino-terminal kinase (JNK) and p38 MAPK. The protective effect of curcumin on P19 induced apoptosis of macrophages were enhanced by treatment with the JNK-specific inhibitors, whereas SB203580, the inhibitor of p38 MAPK had no effect. Curcumin had no effect on the activity of extracellular signal-regulated protein kinase (ERK). Taken together, our data show that the JNK pathway, but not the p38 or ERK pathway, plays an important role in the protective effect of curcumin against P19 induced macrophage apoptosis, and regulation of the JNK pathway may partially elucidate the anti-tuberculosis activity of curcumin.

A prospective multicentre study of mycophenolate mofetil combined with prednisolone as induction therapy in 213 patients with active lupus nephritis.

Mycophenolate mofetil (MMF) with prednisolone has been associated with high remission rates when used as induction treatment for lupus nephritis. This prospective, multicentre, cohort study investigates the efficacy and safety of this regimen over 24 weeks in 213 Chinese patients with active lupus nephritis (Classes III, IV, V or combination). Baseline activity index (AI) was 6.91+/-3.33 and chronicity index (CI) was 1.9+/-1.2. The remission rate was 82.6% at 24 weeks (complete remission, 34.3%; partial remission, 48.4%). There were significant (P<0.01) improvements in kidney function shown by reductions in proteinuria, serum albumin, serum creatinine and creatinine clearance, as well as in systemic lupus erythematosus disease activity index (SLEDAI) scores. Independent risk factors influencing remission were pathological classification (including Class V and III or Class V and IV nephritis) and elevated serum creatinine at baseline (OR 2.967, 95% CI: 1.479-6.332, P=0.001 and OR 1.007, 95% CI: 1.002-1.011, P=0.001, respectively). Patients with concomitant membranous features on biopsy had a lower remission rate than those with Class III and IV nephritis (66.7% vs 87.3%, P=0.002). Renal biopsy was repeated in 25 patients following treatment. There was a transition to less severe pathological morphologies in majority of subjects. Infections were monitored throughout treatment: eight patients (3.8%) experienced bacterial infections, whereas herpes zoster occurred in seven patients. Nine patients (4.2%) suffered from gastrointestinal upset, which resolved without discontinuation of MMF. One patient became leucopenic, whereas another died from active disease unrelated to kidney symptoms. MMF combined with prednisolone is an effective and well-tolerated induction treatment for patients with active lupus nephritis and for controlling SLE systemic activity.