[New progress in the first-line treatment of advanced hepatocellular carcinoma].

Hepatocellular carcinoma is a kind of cancer with a strong invasion, a high incidence rate and mortality, and a poor prognosis. At the time of diagnosis, most patients are already in the advanced stages of a tumor and have lost the chance for radical surgical treatment. Advanced hepatocellular carcinoma treatment has a gradual transition from systemic chemotherapy to targeted therapy, immunotherapy, and combination therapy, especially immune checkpoint inhibitor-based immunotherapy combination therapy, such as combination with bevacizumab monoclonal antibodies and other drugs, or combination with TACE, HAIC, radiotherapy, ablation, and other treatment methods. Combination therapy has significant synergistic effects and thus has already become a future treatment trend for hepatocellular carcinoma. An immunotherapy-based combination therapy plan will run through the whole process of systemic therapy, which is expected to bring better survival benefits to patients with hepatocellular carcinoma. This article reviews the latest research progress in aspects of the first-line treatment of advanced hepatocellular carcinoma.

[Reproducibility evaluation of air-charged catheter for rest urethral pressure profilometry].

We retrospectively included 53 (9 males, 44 females) patients who underwent rest urethral pressure profilometry (RUPP) due to lower urinary tract symptoms (LUTS) in Beijing Chaoyang Hospital from May 2021 to March 2022. The age of patients was (55±16) (25-76) years old. The clinical diagnoses of the included patients with lower urinary tract symptoms were classified as: benign prostatic hyperplasia in 9 cases, interstitial cystitis in 1 case, stress urinary incontinence in 11 cases and non-obstructive dysuria in 32 cases. The full set of urodynamic examination was composed of bladder pressure measurement during the filling period, pressure flow study (PFS) and RUPP. All the urodynamic examination was performed by air-charged catheter (ACC) and corresponding equipment. All patients underwent RUPP measurements twice. The vesical pressure (Pves) values were (35.30±6.15), (35.81±5.91) cmH2O (1 cmH2O=0.098 kPa); the Pura@max were (141.91±36.53), (145.02±38.85) cmH2O; functional urethral length were (41.70±16.34), (42.55±16.40) mm; the maximum urethral closure pressure for the two RUPP measurements were (106.57±36.44), (109.41±39.27) cmH2O. There was no statistical difference between the two RUPP measurements (P>0.05). The reproducibility of the RUPP measurements obtained by ACC is good and deserves further study.

[Diagnostic value of urethral pressure profile for female bladder neck obstruction using air-charged catheters].

Objective: To evaluate the measurements of urethral pressure profile (UPP) using air-charged catheters (ACC) in women with bladder neck obstruction and to preliminarily analyze the diagnostic value of UPP based on ACC for female bladder neck obstruction. Methods: Retrospective inclusion of 63 female patients with bladder neck obstruction diagnosed in the Department of Urology of Beijing Chao-yang Hospital from September 2020 to December 2021 were included as the observation group, and 45 female patients who came to the hospital due to lower urinary tract symptoms with non-bladder neck obstruction during the same period were selected as the control group. All patients received urodynamic examination and UPP examination based on ACC, and urethral pressure measurements [M (Q1, Q3)] were compared between the two groups and analyzed the predictive value of the relevant measurements for female bladder neck obstruction. Results: The age of the observation group was 61 (47, 67) years and body mass index (BMI) was 24.1 (21.6, 27.9) kg/m2, the age of the control group was 65 (58, 71) years and BMI was 25.0 (22.8, 27.6) kg/m2. The urethral pressure at the bladder neck and urinary sphincter position in the observation group were higher than control group[104.00 (83.00, 124.00) cmH2O (1 cmH2O=0.098 kPa) vs 42.00 (31.00, 65.50) cmH2O, 125.0 (107.0, 154.0) cmH2O vs 99.0 (77.0, 124.0) cmH2O, P<0.001]. The results of the receiver operating characteristic (ROC) curve showed that the area under the curve (AUC) of urethral pressure at the bladder neck position had the highest predictive value for female bladder neck obstruction (AUC: 0.946, 95%CI: 0.886-0.981, sensitivity: 87.3%, specificity: 91.1%), and the results of interaction point plot showed that the cut-off value was 72 cmH2O. The cut-off value of urethral pressure at the bladder neck position was used as positive cut-off value to predict female patients with bladder neck obstruction. Sixty-three patients with bladder neck obstruction were detected by UPP measurement using ACC in 59 cases (59/63, 93.7%) and the Kappa value was 0.774 (95%CI: 0.654-0.894), indicating that UPP had high consistency with pressure-flow study. Conclusions: UPP examination based on ACC can show the pressure values of the bladder neck and urethral sphincter position. When the urethral pressure of the bladder neck position is greater than 72 cmH2O, it is helpful for the diagnosis of female bladder neck obstruction and has guiding significance for the diagnosis and treatment of this disease.

Identification and Verification of the Ability of Cdk5 to Phosphorylate Deubiquitinating Enzyme BRCC3 In Vitro.

It is known that the expression of the deubiquitinating enzyme BRCA1-BRCA2-containing complex subunit 3 (BRCC3) and cyclin-dependent protein kinase 5 (Cdk5) is increased in Parkinson's disease (both are involved in neuroinflammatory response). However, the regulatory mechanism of Cdk5 on the post-translational modification of BRCC3 remains unclear. Here we studied whether Cdk5 phosphorylates BRCC3. Phosphorylation of BRCC3 by Cdk5 was predicted by GPS 5.0 software. His-BRCC3 plasmid was constructed by cloning the BRCC3 gene into pGEX-6P-1 vector, and then His-BRCC3 fusion protein was induced with isopropyl ß-d-1-thiogalactopyranoside and purified using His-Tag affinity chromatography purification agarose. Phosphorylation of BRCC3 fusion protein by Cdk5 in vitro was detected by mass spectrometry and Western blotting. The results showed that multiple phosphorylation sites were predicted by GPS 5.0, and the His-BRCC3 fusion protein was successfully induced and purified. In vitro kinase assay, Western blotting, and mass spectrometry showed that Cdk5 can phosphorylate BRCC3. It has been demonstrated that protein kinase Cdk5 can phosphorylate the deubiquitinating enzyme BRCC3 in vitro, which provides new data for further study on the mechanism of neurodegeneration.

Two genetically similar H9N2 influenza viruses isolated from different species show similar virulence in minks but different virulence in mice.

The H9N2 influenza virus has been frequently endemic in poultry, infected mammals and humans and has threatened public health. It is therefore imperative to understand the molecular mechanism enabling this virus to jump from avian to mammalian species. In this study, two H9N2 influenza viruses were isolated from the same region in eastern China but from different hosts; one was isolated from mink and named A/Mink/Shandong/WM01/2014(H9N2)(WM01), while the other was isolated from chicken and named A/Chicken/Shandong/LX830/2014(H9N2)(LX830). Sequencing and phylogenetic analysis showed that both H9N2 influenza viruses had similar genetic backgrounds. The results of infection in minks suggested that both viruses caused significant weight loss and pathological changes in the lungs. Mouse infection showed that LX830 was nonpathogenic in mice, but WM01 resulted in 25% mortality and pathological changes in the lungs, such as severe edema and diffused inflammation of the interalveolar septa. Comparison of the full genomes of both H9N2 influenza viruses showed 52-nucleotide-synonym mutations in 8 gene segments and 7-nucleotide-antonym mutations, resulting in 7 amino acid (AA) substitutions distributed in the PB1, PA, NA and M gene segments. None of these mutations did affect splicing of the M and NS gene segments at the nucleotide level or minor open reading frames (ORFs), such as PB1-F2 and PA-X. Phylogenetic analysis showed that both H9N2 influenza viruses belong to the prevalent epidemic genotype in Asia. Keywords: H9N2 influenza virus; chicken; minks; pathogenicity; phylogenetic.

[AMG-102 inhibits proliferation and induces apoptosis of laryngeal squamous cell carcinoma cells by regulating c-Met/PI3K/Akt pathway].

Objective: To investigate the effects of c-Met inhibitor AMG-102 on the proliferation and apoptosis of laryngeal squamous carcinoma Hep-2 cells and the underlying mechanism. Methods: Laryngeal squamous carcinoma cell line Hep-2 cells were treated with 2.5, 5 and 10 µmol/L AMG-102, respectively. The proliferation activities of Hep-2 cells were detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT). The apoptotic rate of Hep-2 cells was detected by flow cytometry analysis and Hoechst staining. The mRNA expression levels of apoptosis-related genes were detected by real-time quantitative polymerase Chain reaction (RT-qPCR), and the protein expressions of c-Met/PI3K/AKT pathway were detected by western blot. Results: Compared with the control group, the proliferation rates of Hep-2 cells treated with 2.5, 5 and 10 µmol/L AMG-102 for 24 hours were (89.8±1.1)%, (79.8±1.0)% and (69.1±1.2)%, respectively; for 48 hours were (76.8±2.0)%, (60.2±1.1)% and (49.8±1.2)%, respectively; for 72 hours were (50.1±2.0)%, (41.5±1.1)% and (33.6±1.0), respectively, with significant differences (all P<0.05). The apoptotic rates of Hep-2 cells treated with 2.5, 5 and 10 µmol/L AMG-102 for 48 hours were (16.09±1.53)%, (27.51±2.02)% and (36.57±1.42)%, respectively, which were significantly higher than (3.62±0.10) % in the control group (all P<0.05). After treated with 2.5, 5 and 10 µmol/L AMG-102 for 48 hours, the relative expression levels of Bcl-2 mRNA in Hep-2 cells were 0.58±0.13, 0.38±0.12 and 0.20±0.13, respectively; the relative protein expression of p-Met were 80.0±3.8, 50.6±4.2 and 28.5±1.3, respectively; the relative protein expression of p-PI3K were 87.1±0.9, 54.2±1.2 and 21.0±1.2, respectively; the relative protein expression of p-AKT were 98.7±5.6, 56.9±3.2 and 32.2±4.3, respectively; which were significantly lower than those in the control group (all P<0.05). The relative expression levels of Bax mRNA were 1.78±0.13, 2.37±0.14 and 3.05±0.13, respectively, and the relative expression levels of caspase-3 mRNA were 1.98±0.14, 2.47±0.14 and 3.15±0.13, respectively, which were significantly higher than those in the control group (all P<0.05). Conclusion: c-Met inhibitor AMG-102 could inhibit the proliferation and induce apoptosis of laryngeal squamous carcinoma Hep-2 cells by regulating the c-Met/PI3K/Akt pathway.

[Comparison of epidemic characteristics between severe acute respiratory syndrome and coronavirus disease 2019].

Objective: To compare epidemic characteristics between severe acute respiratory syndrome (SARS) and coronavirus disease 2019 (COVID-19). Methods: The general information, including epidemiological and clinical data of the confirmed cases during the epidemic period of the two infectious diseases was collected. The data of SARS in Guangzhou was derived from the technical files of Guangzhou Center for Disease Control and Prevention (GZCDC), including the statistical report and brief report of the epidemic situation (from January 2 to May 11, 2003);The data of COVID-19 in Wuhan was derived from the epidemic data published by Wuhan health and Health Committee's official website and other publicly reported documents (from December 9, 2019 to March 11, 2020). Descriptive analysis was used for a comparativeanalysis of the time and age characteristics, the number of cases, basic reproduction number (R0), proportion of medical staff in confirmed cases (%), crude mortality, etc. Results: A total of 1 072 cases of SARS in Guangzhou were included in the study. The incidence ratio of male to female was 1∶1.26. 43 cases of death were reported with a mortality rate of 4.01%. The median age was 36 years old. The proportion of medical staff in the early stage of the epidemic was 29.04% (88 cases). As to COVID-19 in Wuhan, a total of 49 978 cases were included, The incidence ratio of male to female was 1.04∶1. The 2 423 cases of death were reported with a mortality rate of 4.85%. The median age was 56 years old. The proportion of medical staff in the early stage of the epidemic was 30.43% (42 cases). Conclusion: The COVID-19 in Wuhan has the characteristics of high incidence and wide population. However, the epidemic situation is falling rapidly, and the prevention and control strategy needs to be adjusted timely. The prevention and control of nosocomial infection should be addressed in future.

[Comparison of single incision mini slings and tension-free vaginal tape obturator in the treatment of pure female stress urinary incontinence: results of 2-year investigation].

Objective: To compare the clinical efficacy and safety of single incision mini slings (contasure needless, C-NDL) and tension-free vaginal tape obturator (TVT-O) in the treatment of pure female stress urinary incontinence(SUI). Methods: Clinical data of 107 female SUI patients who underwent C-NDL (n=51) or TVT-O (n=56) procedure during October 2013 and October 2017 in Department of Urology, Beijing Chaoyang Hosptial, Capital Medical University were enrolled.The age of the patients in the C-NDL and TVT-O group was (58.8±9.3) years (range: 43 to 81 years) vs.(56.9±11.4) years (range: 32 to 87 years), the cough leak point pressure was 70(20) cmH(2)O (M(Q(R)), 1 cmH(2)O=0.098 kPa) vs. 80(25) cmH(2)O. The pre- and peri-operative data, subjective cure rate, objective cure rate, quality of life, International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF), Incontinence Impact Questionnaire-Short Form (IIQ-7) and other complications were compared. The t test, Wilcoxon rank sum test, analysis of variance, χ(2) test andFisher exact test were applied to compare the indexes. Results: In the C-NDL and TVT-O group, the median operation time was 30(10) minutes vs. 40(30) minutes (Z=-3.73, P<0.01). The postoperative follow-up period was 32.0(17.4) months vs.41.0(23.2)months. At the latest follow-up, the objective cure rate was 48/51 vs. 54/56 (χ(2)=0.01, P=0.94), the subjective cure rate was 43/51 vs. 50/56 (χ(2)=0.03, P=0.86), ICIQ-SF was 2.29±4.02 (range:0 to 13) vs. 1.71±3.67 (range:0 to 18), and IIQ-7 was 1.12±1.85 (range: 0 to 7) vs.0.70±1.70 (range: 0 to 6). The statistically significant difference in the postoperative visual pain analogue scale (VAS) were found between the two groups (0(1.5) vs.1.0(1.0), Z=3.41, P<0.01). Dysuria occurred in 2 patients in each group, vaginal pain occurred in 1 patient in the C-NDL group, groin pain occurred in 2 patients and urinary urgency occurred in 3 patients in the TVT-O group too. There were no significant difference between the complication rates of the 2 group (3/51 vs.7/51, P=0.325). Conclusion: Both C-NDL and TVT-O may be effective and reliable in the treatment of female SUI.

[Effect of c-Met inhibitor AMG-102 on radiosensitivity in laryngeal squamous carcinoma cells].

Objective: To investigate the effect of c-Met inhibitor AMG-102 on proliferation and radiosensitivity in laryngeal squamous carcinoma cells. Methods: The effects of AMG-102 on proliferation and radiosensitivity of laryngeal squamous carcinoma cell lines Hep-2 and KBV200 were detected by 3-(4, 5-dimethy-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay and colony formation assay, respectively. The apoptosis of Hep-2 and KBV200 cells was detected by flow cytometry. The expression levels of c-Met, phospho-Met (p-Met), cleaved caspase-3 and Akt/p-Akt, Erk/p-Erk were detected by Western blot. Specific small interfering RNA targeting c-Met or plasmid of c-Met were transfected into Hep-2 and KBV200 cells to investigate the cell sensitivity to AMG-102. Results: Compared with KBV200 cells, Hep-2 cells were more sensitive to AMG-102 with IC(50) of 14 and 9 µmol/L, respectively. The relative expression levels of c-Met and p-Met proteins in Hep-2 cells were 194.48±0.57 and 177.76±1.53, respectively, which were significantly higher than those in KBV200 cells (171.24±1.00 and 115.37±0.56, respectively, P<0.001 for both). Exogenous hepatocyte growth factor (HGF) was added to increase the expression level of p-Met protein in KBV200 cells. The results showed that AMG-102 significantly reduced the expression of p-Met in KBV200 cells treated with HGF (P<0.001). Compared with the dimethyl sulfoxide (DMSO) group, AMG-102 treatment combined with radiotherapy significantly increased the radiosensitivity of Hep-2 cells (SER=1.28, P<0.001). However, AMG-102 had little effect on the radiosensitivity of KBV200 cells (SER=1.18, P=0.002). Compared with the 4 Gy radiotherapy alone group and the 5 µmol/L of AMG-102 alone treatment group, the apoptosis rate of Hep-2 cells in the combined treatment group was significantly increased. Meanwhile, the expression level of cleaved caspase-3 protein was also markedly increased. However, there were no significant changes in the apoptotic rate and cleaved caspase-3 expression in each treatment group of KBV200 cells. Compared with DMSO treatment group, the expression levels of p-Met, p-Akt and p-Erk were significantly decreased in the 4 Gy radiotherapy group, 5 µmol/L of AMG-102 treatment group and combined treatment group of Hep-2 cells. And the levels of p-Met, p-Akt and p-Erk in the combined treatment group were significantly lower than those in the 4 Gy radiotherapy alone group and 5 µmol/L of AMG-102 treatment alone group. By contrast, in KBV200 cells, the expression of p-Met, p-Akt and p-Erk in each group was not changed. The relative expression of p-Met in Hep-2 cells before and after radiotherapy at 30 min, 1 h, 4 h, 8 h, 24 h were 99.89±0.61, 138.62±1.00, 163.07±5.00, 87.80±1.85, 90.67±0.65 and 94.09±1.41, respectively. The level of p-Met was slightly increased after radiotherapy at 30 min and 1 h (P<0.001 for all), whereas it was significantly decreased from 4 h to 24 h after radiotherapy (P<0.05 for all). By contrast, the expression of p-Met in KBV200 cells did not change with time after radiotherapy (P>0.05). The sensitivity of Hep-2 cells to AMG-102 was decreased after silencing of c-Met, while the sensitivity of KBV200 cells to AMG-102 was not significantly changed (P>0.05). Moreover, the radiosensitivity of Hep-2 cells in c-Met knockdown group had a slightly increasing trend (SER=1.07, P=0.068). After the treatment with 10 µmol/L of AMG-102, the proliferation rate of c-Met ectopically expressed KBV200 cells was 60.05%±3.23%, It was significantly lower than that of the blank control 90.08%±1.04% and siRNA negative control (90.12%±1.01%, P<0.001). The results suggested that the overexpression of c-Met in KBV200 cells increased the radiosensitivity to AMG-102, whereas depletion of c-Met resulted in resistance to AMG-102 in Hep-2 cells. Furthermore, the radiosensitivity of KBV200 cells that overexpressed c-Met showed a decreased trend (SER=0.7, P=0.005). Conclusions: c-Met inhibitor AMG-102 has a significant inhibitory effect on the proliferation of c-Met overexpressing laryngeal squamous carcinoma cells, leading to increased radiosensitivity. It suggests that molecular targeted therapy against c-Met receptor is more effective in c-Met overexpressed subtype of laryngeal squamous cell carcinoma.

[Application of recurrent neural network in prognosis of peritoneal dialysis].

OBJECTIVE: Deep learning models, including recurrent neural network (RNN) and gated recurrent unit (GRU), were used to construct the clinical prognostic prediction models for peritoneal dialysis (PD) patients based on routine clinical data. The performance of the RNN and GRU were compared with logistic regression (LR), which is commonly used in medical researches. The possible underlining clinical implications based on the result from the GRU model were also investigated. METHODS: We used the clinical data from the PD center of Peking University Third Hospital as the data source. Both the baseline data at the beginning of dialysis, and the follow-up and prognostic data of the patients were used by the RNN and GRU prediction models. The hyper-parameters were tuned based on the 10-fold cross-validation. The risk prediction performance of each model was evaluated via area under the receiver operation characteristic curve (AUROC), recall rate and F1-score on the testset. RESULTS: A total of 656 patients with the 261 occurrences of death were included in the experiment. The total number of all diagnostic records were 13 091. The results on the testset showed that the AUROC of the LR model, RNN model, and GRU model was 0.701 4, 0.786 0, and 0.814 7, respectively. The predictive performances of the GRU and RNN models were significantly better than that of the LR model. The performances of the GRU and RNN models assessed by recall rate and F1-score were also significantly better than that of the LR model, in which the GRU model reached the best performance. In addition, the recall rates were different among different causes of death or by different prediction time windows. CONCLUSION: The recurrent neural network model, especially the GRU model, is more effective in predicting PD patients' prognosis as compared with the LR model. This new model may be helpful for clinicians to provide timely intervention, thus improving the quality of care of PD.

Videolaryngoscopy vs. Macintosh laryngoscopy for double-lumen tube intubation in thoracic surgery: a systematic review and meta-analysis.

Double-lumen intubation is more difficult than single-lumen tracheal intubation. Videolaryngoscopes have many advantages in airway management. However, the advantages of videolaryngoscopy for intubation with a double-lumen tube remain controversial compared with traditional Macintosh laryngoscopy. In this study, we searched MEDLINE, Embase, Cochrane Library and the Web of Science for randomised controlled trials comparing videolaryngoscopy with Macintosh laryngoscopy for double-lumen tube intubation. We found that videolaryngoscopy provided a higher success rate at first attempt for double-lumen tube intubation, with an odds ratio (95%CI) of 2.77 (1.92-4.00) (12 studies, 1215 patients, moderate-quality evidence, p < 0.00001), as well as a lower incidence of oral, mucosal or dental injuries during double-lumen tube intubation, odds ratio (95%CI) 0.36 (0.15-0.85) (11 studies, 1145 patients, low-quality evidence, p = 0.02), and for postoperative sore throat, odds ratio (95%CI) 0.54 (0.36-0.81) (7 studies, 561 patients, moderate-quality evidence, p = 0.003), compared with Macintosh laryngoscopy. There were no significant differences in intubation time, with a standardised mean difference (95%CI) of -0.10 (-0.62 to 0.42) (14 studies, 1310 patients, very low-quality evidence, p = 0.71); and the incidence of postoperative voice change, odds ratio (95%CI) 0.53 (0.21-1.31) (7 studies, 535 patients, low-quality evidence, p = 0.17). Videolaryngoscopy led to a higher incidence of malpositioned double-lumen tube, with an odds ratio (95%CI) of 2.23 (1.10-4.52) (six studies, 487 patients, moderate-quality evidence, p = 0.03).

Peroxisome proliferator-activated receptor delta regulates lipid droplet formation and transport in goat mammary epithelial cells.

Even though recent evidence in goat mammary epithelial cells (GMEC) suggest a role of peroxisome proliferator-activated receptor delta (PPARD) in regulating lipid homeostasis, its role is not fully understood. Our hypothesis was that PPARD regulates lipid transport processes in GMEC and, thus, plays a crucial role in regulating fat formation. The PPARD was overexpressed using an adenovirus system (Ad-PPARD) with recombinant green fluorescent protein (Ad-GFP) as the control. Results revealed that overexpression of PPARD markedly upregulated the mRNA abundance of PPARD. Compared with the control (Ad-GFP+dimethyl sulfoxide), overexpression of PPARD alone had no effect on mRNA expression of CD36, SCD1, FABP4, ACSL1, and ADRP. The cultures overexpressing PPARD with the PPARD ligand GW0742 (GW) upregulated the expression of CD36, FABP3, FABP4, ACSL1, and ADRP. Overexpression of PPARD in GMEC plus GW increased the concentration of 16:1 and 18:1-trans and was associated with upregulation of SCD1. Compared with the control (Ad-GFP+dimethyl sulfoxide), the decrease of triacylglycerol concentration coupled with upregulation of genes related to lipid droplet secretion (e.g., ADRP and ACSL1) induced by PPARD overexpression suggests a role in lipid droplet (LD) secretion. Luciferase assay revealed that GW increased the ADRP promoter activity in a dose-dependent manner. Knockdown of PPARD impaired the increase of ADRP promoter activity induced by GW, whereas GW enhanced the activity of ADRP promoter in GMEC overexpressing PPARD. Data with the ADRP 5'-flanking truncated luciferase reporter suggest a core region (-1,444 to -990 bp) response element for the induction of GW. This core region contains a known PPARG response element (PPRE) at -1,003 to -990 bp. When the PPRE was mutated, the overexpression of PPARD had no effect on ADRP promoter activity. Collectively, these results reveal a novel role for PPARD in lipid homeostasis via promoting fatty acid transport and LD formation through a mechanism of direct binding to the promoter of key genes. Hence, PPARD activity may contribute to fatty acid transport and LD formation during lactation.

Fatty acid elongase 5 (ELOVL5) alters the synthesis of long-chain unsaturated fatty acids in goat mammary epithelial cells.

Increased production of long-chain unsaturated fatty acids (LCUFA) can have a positive effect on the nutritional value of ruminant milk for human consumption. In nonruminant species, fatty acid elongase 5 (ELOVL5) is a key enzyme for endogenous synthesis of long-chain unsaturated fatty acids. However, whether ELOVL5 protein plays a role (if any) in ruminant mammary tissue remains unclear. In the present study, we assessed the mRNA abundance of ELOVL5 at 3 stages of lactation in goat mammary tissue. Results revealed that ELOVL5 had the lowest expression at peak lactation compared with the nonlactating and late-lactating periods. The ELOVL5 was overexpressed or knocked down to assess its role in goat mammary epithelial cells. Results revealed that ELOVL5 overexpression increased the expression of perilipin2 (PLIN2) and decreased diacylglycerolacyltransferase 2 (DGAT2) and fatty acid desaturase 2 (FADS2) mRNA, but had no effect on the expression of DGAT1, FADS1, and stearoyl-CoA desaturase 1 (SCD1). Overexpression of ELOVL5 decreased the concentration of C16:1n-7, whereas no significant change in C18:1n-7 and C18:1n-9 was observed. Knockdown of ELOVL5 decreased the expression of PLIN2 but had no effect on DGAT1, DGAT2, FADS1, FADS2, and SCD1 mRNA expression. Knockdown of ELOVL5 increased the concentration of C16:1n-7 and decreased that of C18:1n-7. The alterations of expression of genes related to lipid metabolism after overexpression or knockdown of ELOVL5 suggested a negative feedback regulation by the products of ELOVL5 activation. However, the content of triacylglycerol was not altered by knockdown or overexpression of ELOVL5 in goat mammary epithelial cells, which might have been due to the insufficient availability of substrate in vitro. Collectively, these are the first in vitro results highlighting an important role of ELOVL5 in the elongation of 16-carbon to 18-carbon unsaturated fatty acids in ruminant mammary cells.

A method for improving the accuracy of non-invasive prenatal screening by cell-free foetal DNA size selection.

Background Non-invasive prenatal screening (NIPS) using cell-free foetal DNA (cfDNA) has been widely used for identifying common foetal aneuploidies (e.g. trisomy 21 (T21), trisomy (T18) and trisomy 13 (T13)) in clinical practice. The sensitivity and specificity of NIPS exceeds 99%, but the positive prediction value (PPV) is approximately 70% (combined T21, T18 and T13). Thus, some 30% of pregnant women who have positive NIPS results are eventually identified as normal by amniocentesis. These women therefore must undertake needless invasive tests and risk miscarrying healthy babies because of false positive NIPS results. Methods In order to achieve higher accuracy, we amended the standard NIPS (s-NIPS) protocol with an additional cfDNA size selecting step in agarose-electrophoresis. The advantage of the new method (named e-NIPS) was validated by comparing the results of e-NIPS and s-NIPS using 114 retrospective cases selected from 15,930 cases. Results Our results showed that the foetal cfDNA fraction can be enriched significantly by a size selection step. With this modification, all 98 negative cases and 9 of 11 false positive cases of s-NIPS were correctly identified by e-NIPS, resulting in an increased PPV from 71% to 77%. Additionally, a simulation test showed that e-NIPS is more reliable than s-NIPS, especially when the foetal cfDNA concentration and sequencing coverage are low. Conclusion cfDNA size selection is an important step in improving the accuracy of non-invasive prenatal screening for chromosomal abnormalities.

[Curative effects of Ningmitai capsules on the residual fragments and postoperative complications following lithotripsy treatment of upper urinary calculi].

Objective: To observe the clinical efficacy of Ningmitai capsules, a traditional Chinese medicine using for clearing heat and dampness, in the treatment of residual fragments and postoperative complications following lithotripsy for upper urinary stones. Methods: During October 2016 and March 2018, patients from Wuhan 1st Hospital, Wuhan 2nd Hospital, Wuhan 3rd Hospital, and Wuhan Puai Hospital having upper urinary residual fragments following minimally-invasive stone treatment were randomly assigned to control group and Ningmitai group with a proportion of 1∶3. The patients in control group were treated with antibiotics or sodium diclofenac suppository on demand, while patients in Ningmitai group took additional Ningmitai capsule orally (4 capsules per time, 3 times per day). The observation was started when a patient was enrolled in this study and continued for a maximum of 12 weeks or until stone-free status. During the observation, the stone expulsion time, stone-free time, stone-free rate were observed, and the difference in curative effect between the two groups on postoperative complications such as pain and infection were compared. Statistical analysis was done using t-test or χ2 test by GraphPad Prism 5 software. Results: Totally 269 cases enrolled in this study. Eighty-six patients were from Wuhan 1st Hospital, 69 patients from Wuhan 2nd Hospital, 58 patients from Wuhan 3rd Hospital, 56 patients from Wuhan Puai Hospital, respectively. There were 66 cases in control group and 203 cases in Ningmitai group. The residual fragments expulsion time in Ningmitai group was significantly earlier than that in control group ((4.5±0.4) days vs. (7.5±1.3) days, t=2.877, P=0.004), the residual fragments clearance time in Ningmitai group was significantly shorter than that in control group ((13.6±1.0) days vs.(25.6±3.8) days, t=4.252, P=0.000), and the stone-free rate within 4 weeks post operation in Ningmitai group was significantly higher than control group (91.6% vs. 68.2%, χ2=22.57, P=0.000). After 12 weeks of treatment, the total effective rate of control group was 89.4%, and the total effective rate of Ningmitai group was 99.5%, with statistically significant difference (χ2=17.65, P=0.000). The proportion of caregivers that offered analgesia in Ningmitai group was significantly lower than that in control group (16.3% vs. 30.3%, χ2=6.212, P=0.013), the recovery rate of routine urinalysis following 4 weeks' treatment was significantly higher in Ningmitai group than that in control group (88.2% vs.71.2%, χ2=10.67, P=0.001). No obvious adverse effects were observed in both groups. Conclusions: Ningmitai capsule can facilitate the stone passage and increase the stone-free rate in the treatment of residual fragments and postoperative complications of upper urinary stones. It is also helpful for the prevention and treatment of postoperative pain, infection and other complications.

[Epidemiological investigation on the incidence of heat stroke in Huzhou from 2013 to 2017].

Objective: To understand the status of heat stroke in Huzhou and take preventive strategy and measures against the heat stroke. Methods: Case reports between 2013 and 2017 were collected. The differents between the population and age data from different types of heat stroke were tested by F test, and the genders tested by χ(2) test. The difference was statistically significant (P< 0.05) . Statistical significance by R 3.4.1. Results: A total of 1 307 patients were reported from 2013 to 2017, among which 878 cases (67.18%) were mild and 429 cases (32.82%) were severe. 15 fatalities (1.15%) were from the severe heat stroke cases. Male patients (947 cases, 72.46%) were more than female ones (360 cases, 27.54%) and the difference was statistical significant (χ(2)=8.541, P<0.05) . Heat stroke cases in age 45~55 (25.33%) accounted for the largest proportion and the difference was statistical significant (F=8.87, P<0.05) . The cases for heat stroke have increased year by year and have obvious seasonality, mainly in June to September. Conclusion: Taken together, these findings indicated that the incidence rate of heat stroke were mainly mild and middle-aged men were susceptible to heat stroke in the city. However, to decrease the numbers of heat stroke, the relevant departments should take preventive strategy and measures against the heat stroke.

[Applied study of Singapore occupational semi-quantitative risk assessment technique in occupational health risk assessment in a chair furniture manufacturing enterprise].

Objective: To assess the occupational health risk level of a small-scale furniture manufacturer, and to explore the applicability of the Singapore-developed semi-quantitative occupational risk assessment model for chemical exposure (Singapore model) in small-scale furniture manufacturers, and to provide a basis for the continuous occupational health management of manufacturers. Methods: A small-scale furniture manufacturer was selected as the study subject; an on-site occupational hygiene investigation was performed on the above manufacturer during April to June in 2017, and a risk assessment was carried out using the Singapore model. Results: The assessment results of the Singapore model indicated that risk levels of occupational exposure to harmful chemicals for the key positions in the workplace were inconsistent between the actual exposure level method and the exposure index method except for the following: high risk for formaldehyde exposure (risk level: 3.5 and 4.1, respectively) during woodworking process, high risk for dimethyl benzene exposure (risk level: 3.5 and 3.5, respectively) during burnishing process, medium risk for methyl benzene and dimethyl benzene exposure (risk level: 3.0 and 3.4, respectively) as well as for dimethyl benzene exposure (risk level: 3.0 and 3.3, respectively) during primer coating process, medium risk for methyl benzene exposure (risk level: 3.0 and 3.4, respectively) during gel painting process, and medium risk for cyclohexanone exposure (risk level: 2.8 and 2.8, respectively) during oil polishing process. The exposure index method yielded a higher risk level than the actual exposure level method, especially in the risk level of benzene exposure, which was rated as "high" and "very high" by the former but "low" by the latter. Conclusion: The Singapore model is suitable for risk assessment of occupational exposure to harmful chemicals in small-scale furniture manufacturers, which can provide a basis for further prevention and control measures taken by manufacturers.

[Effect of occupational stress on recurrent spontaneous abortion in women of childbearing age].

Objective: To investigate the influence of occupational stress on recurrent spontaneous abortion (RSA) in women of childbearing age. Methods: From January to December, 2017, 75 working women of childbearing age (25-35 years) who were admitted to a provisional hospital in Lanzhou, China and diagnosed with RSA were assigned into patient group. At a 1∶4 ratio, 300 age-matched working women who had normal first pregnancy were randomly selected as controls. A case-control study was conducted by a self-made questionnaire and the effort-reward imbalance scale. The impact of occupational stress on RSA in women of childbearing age was analyzed by evaluation of occupational harmful factors, regularity, effort-reward ratio, and sleep quality. Results: There were significant differences in the distribution of sleep, daily exercise, night shift, extrinsic-effort/low-reward score, and effort/low-reward score between the patient group and the control group (χ(2)=7.867, P<0.05; χ(2)=7.377, P<0.05; χ(2)=3.714, P<0.05; χ(2)=6.651, P<0.05; χ(2)=8.556, P<0.05) . With controlled factors such as general conditions and living habits, logistic regression analysis showed that poor sleep quality and high-effort/low-reward were risk factors for RSA (odds ratio[OR]=1.462, 95% confidence interval[CI]: 1.032~2.073; OR=3.253, 95%CI: 1.169~9.053) . A regular work was a protective factor against RSA (OR=0.644, 95%CI: 0.438-0.946) . Conclusion: In occupational stress, irregular working hours, lack of sleep, and high-effort/low-reward are risk factors for RSA. Working women of childbearing age should ensure adequate sleep, pay attention to effort-reward balance, and make a regular work schedule.

Identification and characterization of Laodelphax striatellus (Insecta: Hemiptera: Delphacidae) neutral sphingomyelinase.

The neutral sphingomyelinase (nSMase) 1 homologue gene LsSMase was cloned from Laodelphax striatellus, a direct sap-sucker and virus vector of gramineous plants, and expressed via a Bac to Bac baculovirus expression system. The LsSMase-enhanced green fluorescent protein fusion protein was located in the endoplasmic reticulum in a similar manner to mammalian nSMase 1. The biochemical properties of LsSMase were determined in detail. The optimal pH and temperature for recombinant LsSMase were 8 and 37 °C, respectively. LsSMase was an Mg2+ or Mn2+ dependent enzyme, but different concentration of each were needed. The activity of LsSMase was significantly stimulated by Ethylene glycol bis(2-aminoethyl ether)tetraacetic acid (EGTA), whereas it was inhibited by ethylenediaminetetraacetic acid. Millimolar concentrations of Zn2+ completely inhibited LsSMase. The reducing agents dithiothreitol and ß-mercaptoethanol varied in their effects on activity. Phospholipids were not found to stimulate LsSMase.