Objective To establish an HPLC method to determine the concentration of inositol nicotinate（IN） in rat skin, and study the pharmacokinetic characteristics of IN after transdermal administration of heparin sodium inositol nicotinate cream in rats. Methods HPLC method was used to establish a simple and rapid analytical method for the determination of IN concentration in the skin of rats at different time points after administration. The established method was used to study the pharmacokinetics of IN after transdermal administration of heparin sodium inositol nicotinate cream in rats, and the pharmacokinetic parameters were fitted with DAS software. Results The linearity of the analytical method was good in the concentration range of 0.25-20 μg/ml, the quantitative limit was 0.25 μg/ml, and the average recovery rate was 96.18%. The pharmacokinetic parameters of IN after transdermal administration of heparin sodium inositol nicotinate cream in rats were as follows: t_1/2 was （4.555±2.054） h, T_max was （6±0）h, C_max was （16.929±2.153）mg/L, AUC_0−t was （150.665±16.568） mg·h /L ,AUC_0−∞ was （161.074±23.917） mg·h /L, MRT_（0−t） was （9.044±0.618）h, MRT_{（0−∞）} was （10.444±1.91） h, CLz/F was （0.19±0.03） L/（h·kg）, and Vz/F was （1.19±0.437） L/（h·kg）. Conclusion IN could quickly penetrate the skin and accumulate in the skin for a long time, which was beneficial to the pharmacological action of drugs on the lesion site for a long time. The method is simple, rapid, specific and reproducible, which could be successfully applied to the pharmacokinetic study of IN after transdermal administration in rats.

Objective:To analyze the Chinese thyroid imaging reporting and data system (C-TIRADS) classification combined with ultrasonic shear wave elastography (SWE) and super microvascular imaging technology (SMI) in the diagnosis of benign and malignant thyroid nodules.Methods:Clinical data of 125 patients undergoing thyroid nodule surgery in the Department of Ultrasound Medicine, Shantou Central Hospital were retrospectively analyzed. There were 35 males and 90 females. The disease duration was (3.45±1.32) years, ranging from 3 months to 7 years; the age was (55.45±3.31) years old, ranging from 25 to 70 years old; the maximum diameter of the nodule is (12.13±5.76) mm, ranging from 5.0 to 42.9 mm. C-TIRADS classification, SWE, SMI and pathological diagnosis results of ultrasonic nodules in patients were analyzed. The diagnostic performance of C-TIRADS classification, SWE, SMI and combined diagnosis of malignant nodules were analyzed. SPSS 21.0 software was used to analyze the data, and the measurement data were consistent with the normal Statistical distribution, independent t test was performed for comparison between two groups; count data were expressed as rate (%), and χ2 test was used. Results:A total of 180 lesions were detected this time. According to pathological diagnosis, there were 114 benign nodules and 66 malignant nodules; the malignant percentage of C-TIRADS 4B nodules was the highest, reaching 72.00% (36/50) ; Emix of malignant nodules, Emax and Emean were (15.98±6.56) kPa, (84.22±24.23) kPa and (63.29±15.89) kPa respectively, which were significantly higher than those of benign nodules (13.56±4.68) kPa, (48.33±14.46) kPa and (37.32±12.63kPa) ( t=2.49, 8.76, 9.07, P<0.05) ; Type IV malignancy accounted for the highest proportion in SMI classification of thyroid nodules, which was 78.95%; the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of the joint diagnosis were 94.64%, 92.85%, 94.64%, 42.85%, 90.16%, higher than the single C-TIRADS classification (89.65%, 75.00%, 89.65%, 37.50%, 80.55%), SMI (82.35%, 55.56%, 89.36%, 32.60%, 78.33%), and SWE diagnosis (81.08%, 56.00%, 90.22%, 28.57%, 77.22%). The difference was statistically significant ( P<0.05) . Conclusions:Compared with the diagnostic efficiency of SWE, SMI and C-TIRADS classification alone, the combined use of the three has higher diagnostic efficiency for benign and malignant thyroid nodules. The abuse of invasive procedures can be reduced or avoided.

Objective To investigate the role of sodium-glucose cotransporter 1(SGLT1)inhibitor mizagliflozin(MIZA)in autosomal dominant polycystic kidney disease(ADPKD).Methods Western blotting,quantitative polymerase chain reaction(qPCR),and immunofluorescence staining were used to determine the expression and distribution of SGLT1 in kidney tissues of PKD1-/-and PKD1+/+mice,human renal cancer adjacent tissue and ADPKD tissue.Renal cyst lining epithelial cells OX161 and renal tubular epithelial cells UCL93 were treated with MIZA,incubated at 37℃for 24,48,and 72 h,and then were subjected to methyl thiazolyl tetrazolium and colony formation assay to observe cell proliferation.The qPCR method was used to determine the mRNA levels of collagen 1α1,collagen 3α1,and fibronectin 1 in OX161 cells treated with 100 μmol/L MIZA for 48 h.The Madin-Darby canine kidney(MDCK)cell 3D cyst formation assay verified the effect of MIZA on cyst formation.The mRNA-seq technology was used to detect differentially expressed genes between UCL93 cells and OX161 cells,and between OX161 cells and OX161 cells treated with 100 μmol/L MIZA for 48 h,and then the differentially expressed genes were analyzed with Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Results The expression level of SGLT1 was significantly increased in the tissues of ADPKD patients and PKD1-/-mice compared to those in normal kidney tissues(P＜0.05,P＜0.01).Immunofluorescence staining revealed that SGLT1 was mainly expressed in the cystic lining epithelial cells.Additionally,MIZA inhibited the proliferation and fibrosis of polycystic kidney cells in a concentration-and time-dependent manner,and also inhibited cyst formation in 3D formation assay in vitro.The mRNA-seq analysis and KEGG enrichment analysis showed that differentially expressed genes between OX161 cells and OX161 cells cultured in 100 μmol/L MIZA for 48 h were mainly enriched in the phosphatidylinositol 3-kinase(PI3K)-protein kinase B(Akt)and mitogen-activated protein kinase(MAPK)signaling pathways,which were the same as those between OX161 cells and UCL93 cells.Conclusion The SGLT1 inhibitor MIZA may inhibit the proliferation and fibrosis of polycystic kidney cells through signaling pathways such as PI3K-Akt and MAPK,delaying the growth of polycystic kidney,and it is a potential therapeutic target for ADPKD.

Objective To evaluate the value of high-frequency color ultrasound for different types of thyroid nodules during healthy examinations. Methods A total of 150 subjects with ultrasound diagnosis of thyroid nodule abnormality were enrolled. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of ultrasound diagnosis for differentiating benign and malignant thyroid nodules were evaluated with histopathological examinations of surgical thyroid specimens as a gold standard. Results A single thyroid nodule was found in all patients, and histopathological examinations identified 102 benign thyroid nodules and 48 malignant nodules. Microcalcification showed the highest accuracy for prediction of benign and malignant thyroid nodules (98.1%; χ² = 45.67, P = 0.001), followed by taller than wider shape of thyroid nodule (92.1%; χ² = 34.06, P = 0.001), central vascularity (82.0%; χ² = 13.29, P = 0.001), halo (76.0%; χ² = 6.15, P = 0.008) and hypoechogenicity (70.3%; χ² = 10.63, P = 0.001). In addition, ultrasound diagnosis showed a 100.0% sensitivity, a 94.1% specificity, a 88.9% positive predictive value, a 100.0% negative predictive value and a 96.0% accuracy for differentiation between benign and malignant thyroid nodules (χ² = 41.830, P < 0.001). Conclusion High-frequency color ultrasound has a high diagnostic accuracy for differentiating benign and malignant thyroid nodules, and microcalcification and taller than wider shape of thyroid nodule exhibit high values for predicting the malignancy of thyroid nodules.

Objective : To discuss and summarize the preventive measures and treatment methods for aspiration/ingestion during dental procedures. Methods : One case of aspiration during an implant operation was reported, and the literature on aspiration/ingestion during oral procedures was reviewed. Results: An implant screwdriver accidentally slipped into the mouth of the patient during implant surgery. The patient experienced no obvious discomfort except a few coughs. The surgeon and assistant paused the procedure immediately to search for the screwdriver, but it was not found. The patient declared that there was no special abnormality, such as breathing disorder or chest distress, so we considered that the foreign body was ingestion. After the implant surgery was completed, no foreign body was found in the stomach via gastroscopy. Chest X-ray and CT showed a dense metal shadow in the lower lobe of the left lung. Under local anesthesia, bronchoscopy and biopsy forceps were used by respiratory physicians to clip out the foreign body. After removal of the foreign body, the patient had no obvious discomfort but a slight cough. Cephalexin and metronidazole were given for three days to prevent infection. Three days later, the patient had no complaints of respiratory discomfort. After reviewing the literature, we found that the operation should be paused immediately after aspiration/ingestion occurs during dental procedures and that the dental chair should be laid down to prevent the foreign body from descending deeper, which may increase the difficulty of removal and cause gastrointestinal and respiratory tract injury. The position of the foreign body should be determined by imaging examination, and the corresponding means to remove the foreign body should be performed. Conclusion Patients may have no obvious symptoms after aspiration/ingestion during dental procedures, and the foreign body can be removed after imaging examination.

Background: Xylazole (Xyl) is a veterinary anesthetic that is structurally and functionally similar to xylazine. However, the effects of Xyl in vitro remain unknown. Objectives: This study aimed to investigate the anesthetic mechanism of Xyl using fetal rat nerve cells treated with Xyl. Methods: Fetal rat nerve cells cultured for seven days were treated with 10, 20, 30, and 40 μg/ mL Xyl for 0, 5, 10, 15, 20, 25, 30, 45, 60, 90, and 120 min. Variations of amino acid neurotransmitters (AANTs), Nitric oxide-Cyclic GMP (NO-cGMP) signaling pathway, and ATPase were evaluated. Results: Xyl decreased the levels of cGMP and NO in nerve cells. Furthermore, Xyl affected the AANT content and Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase activity in nerve cells. These findings suggested that Xyl inhibited the NO-cGMP signaling pathway in nerve cells in vitro. Conclusions This study provided new evidence that the anesthetic and analgesic effects of Xyl are related to the inhibition of the NO-cGMP signaling pathway.

Objective:To compare the laboratory diagnostic methods of Mycoplasma pneumonia(MP) and evaluate its clinical value.Methods:A prospective study.Throat swabs and double sera of children with MP infection were collected from December 2016 to January 2017 in Shengjing Hospital Affiliated to China Medical University; throat swab samples of healthy children aged 3 to 5 in Chaoyang District, Beijing were collected from March to May 2017.Passive agglutination (PA) was used to detect the double serum.Taking the 4-fold increase or decrease of the specific antibody titer of the double serum as the gold standard, the receiver operating characteristic curve (ROC) was drawn, and the laboratory methods for detecting MP infection were compared and evaluated.Results:(1)A total of 93 children with MP infection were clinically diagnosed, including 42 males (45.2%) and 51 females (54.8%), with an average age of 5.5 years.Sixty cases (64.5%) of MP infection were diagnosed.There were 349 healthy children, 198 males and 151 females, with an average age of 4.3 years.The positive rate of throat swab culture was 0.6% (2 cases), and the positive rate of fluorescent quantitative PCR(qPCR) was 18.9% (66 cases). (2) The culture specificity was the highest (100.0%) and the sensitivity was the lowest (65.0%). PA and enzyme linked immunosorbent assay (ELISA) were used to detect a single serum in the acute phase, the sensitivity was 71.7% and 86.5% respectively.ROC curve suggested that the current clinical diagnostic threshold MP specific antibody IgM ≥ 1∶160 was not the best diagnostic threshold.Molecular biological diagnostic methods were the most sensitive, RNA simultaneous and testing (SAT) was 85.0% and qPCR was 93.0%; while the specificity was low, 75.7% (SAT) and 63.6% (qPCR), respectively.(3) At the same time, MP nucleic acid (SAT, PCR) of throat swabs and a single serum (ELISA, PA) of children in acute phase were detected, the sensitivity was increased to 95.0%-100.0%, and the specificity was 63.6%-75.7%.Conclusions:Molecular biology is highly sensitive in diagnosing MP infection.It has asymptomatic infection or is carried after infection.Whether it needs treatment needs to be combined with clinical practice, when MP detection is positive.The detection of a single serum in the acute phase with a course of about 1 week has high sensitivity and is of reference value for the diagnosis of MP infection, but the diagnosis needs to be combined with clinical practice.The sensitivity and accuracy of detecting MP infection by single serological test combined with SAT in acute phase are higher than that by single application.

OBJECTIVE: To analyze the clinical manifestations of heart, liver and kidney damages in the early stage of COVID-19 to identify the indicators for these damages. METHODS: We analyzed the clinical features, underlying diseases, and indicators of infection in 12 patients with COVID-19 on the second day after their admission to our hospital between January 20 and February 20, 2020.The data including CK-MB, aTnI, BNP, heart rate, changes in ECG, LVEF (%), left ventricular general longitudinal strain (GLS, measured by color Doppler ultrasound) were collected.The changes of liver function biochemical indicators were dynamically reviewed.BUN, UCR, eGFR, Ccr, and UACR and the levels of MA, A1M, IGU, and TRU were recorded. RESULTS: The 12 patients included 2 severe cases, 8 common type cases, and 2 mild cases.Four of the patients presented with sinus tachycardia, ECG changes and abnormal GLS in spite of normal aTNI and LVEF; 1 patient had abnormal CKMB and BNP.On the first and third days following admission, the patients had normal ALT, AST and GGT levels.On day 7, hepatic function damage occurred in the severe cases, manifested by elevated ALT and AST levels.Abnormalities of eGFR, Ccr and UACR occurred in 8, 5 and 5 of the patients, respectively.Abnormal elevations of MA, A1M, IGU and TRU in urine protein were observed in 4, 4, 5, and 2 of the patients, respectively. CONCLUSIONS In patients with COVID-19, heart damage can be identified early by observing the GLS and new abnormalities on ECG in spite of normal aTNI and LVEF.Early liver injury is not obvious in these patients, but dynamic monitoring of the indicators of should be emplemented, especially in severe cases. In cases with normal CR and BUN, kidney damage can be detected early by calculating eGFR, Ccr and UACR and urine protein tests.
Betacoronavirus ; COVID-19 ; Coronavirus Infections ; Humans ; Pandemics ; Pneumonia, Viral ; SARS-CoV-2

Objective:To investigate the effects of poly (ADP-ribose) polymerase-1(PARP-1) in intestinal mucosal barrier injury in rat model with severe acute pancreatitis (SAP).Methods:Twenty healthy male Wistar rats were divided into four groups ( n=5 each group) using a random table method: control, SAP, 3-aminobenzamide (3-AB), and 3-AB control groups. The SAP model was induced by intraperitoneal injection of cerulean with lipopolysaccharide. At 30 min, the rats were treated with the PARP-1 inhibitor, 3-AB, or normal saline,separately. After 12 h, all rats were sacrificed to harvest pancreas tissues, intestines tissues, and blood from the hearts for index detection. Serum amylase (AMY) and interleukin (IL)-6 levels were measured using an automatic biochemical instrument and enzyme-linked immunosorbent assay (ELISA), respectively.The protein expression of PARP-1 and nuclear factor (NF-κB) were measured using Western blot and that of occludin was measured using an immunohistochemical test. One-way analysis of variance was used for comparison of multiple groups of variables. Non-parametric tests of rank conversion were used when variances were not uniform. A P <0.05 was considered statistically significant. Results:Compared to the control group, the following indexes in the SAP group were significantly increased: ascites (with serious hemorrhage and necrosis in the pancreas and disordered intestinal villi),serum AMY and IL-6 levels, and the expression of PARP-1 and NF-κB. However, Occludin expression was significantly decreased. There was no significant difference between 3-AB group and 3-AB control group. Compared to the SAP group, the severity of SAP and pancreatitis-associated intestinal injury was significantly attenuated with the administration of 3-AB. Serum AMY and IL-6 levels were significantly decreased (serum AMY: 1 879.25 ± 736.6 U/L vs 5 569.33 ± 1993.48 U/L; IL-6: 77.98 ± 20.65 pg/mL vs 209.14 ± 79.08 pg/mL, both P<0.05), but the expression of PARP-1 and NF-κB were significantly increased (PARP-1: 1.44 ± 0.09 vs 1.49 ± 0.13; NF-κB: 0.63 ± 0.09 vs 0.96±0.08, both P<0.05). Similarly, Occludin expression was significantly decreased (6.7±1.5 vs 3.2±1.1, P<0.05). Conclusions:Inhibition of PARP-1 has protective effects on SAP associated intestinal mucosal barrier damage. The mechanism may be related to the inhibition of NF-κB signaling pathway and increase intestinal mucosal Occludin protein expression.

Objective:To verify the value of whole genomic copy number variation (WGCNV) detection and scoring system in the diagnosis and prognosis of lung adenocarcinoma.Methods:Seventy-six lung adenocarcinoma specimens including ninety-one tumor samples and twenty adjacent non-tumor lung tissue samples were collected using Laser capture microdissection (LCM). Whole genomic amplification (WGA) was used to enrich DNA and construct a sequencing library for next generation sequencing (NGS). Changes of larger than 5Mb CNV in this study were analyzed and scored. The nuclear grading and score of tumor cells in the surgery and pleural effusion cytology of lung adenocarcinoma specimens were evaluated separately. For each case, we evaluated (1) nuclear size, (2) mitotic counts, (3) nuclear atypia, (4) atypical mitoses. The data of disease-free survive (DFS) and overall survive (OS) were collected for assessing the prognostic value of WGCNV score. Meanwhile, receiver operating characteristic (ROC) and area under curve (AUC) were used to define a cut-off value and evaluate the diagnostic significance in lung adenocarcinoma.Results:The WGCNV scores of twenty adjacent non-tumor lung tissue samples were treated as normal control and all of WGCNV scores of tumor samples range from 0 to 9.95, the median score was 2.7. The WGCNV scores were divided into three groups: low score group <1.74, medium score grade 1.74~4.23, high score grade >4.23. The WGCNV score was positively associated with the nuclear grade scoring ( r=0.780 90, P<0.001). The result for evaluation of prognostic value of the WGCNV scores showed that comparing with low WGCNV score group, Hazard Ratio (HR) of medium score group was 4.11 (95% CI=0.72~23.57) and high score group was 2.07 (95% CI=0.30~14.12). These results suggested that the risks of the medium and high WGCNV score group elevated. According to the analysis results of ROC curve, when the cut off value was 0.01, the sensitivity and specificity for lung adenocarcinoma diagnosis were 97.8% and 95.0% respectively, the positive predictive value (PPV) and negative predictive value (NPV) were 99.0% and 90.1%, respectively, the AUC was 0.981. In the differentiation of adenocarcinoma in situ (AIS) and minimally invasive adenocarcinoma (MIA) group and invasive adenocarcinoma group, when the cut off value was 1.8, the sensitivity and specificity between the two groups were 78.1% and 94.4%, and the PPV and NPV were 98.0% and 52.0%, respectively, the AUC was 0.896. Conclusion:This study verifies that WGCNV scoring system has a potential diagnostic and prognostic value in lung adenocarcinoma.