[Mechanism of piRNA in bisphenol A-promoted invasion and migration of prostate cancer cells].

Objective: To investigate the regulatory mechanisms of piwi-interacting RNA (piRNA) in bisphenol A (BPA)-induced prostate cancer cell invasion and migration. Methods: The Cancer Genome Atlas (TCGA) data was used to analyze and screen for piRNAs with significantly increased expression in prostate cancer tissues. PC-3 cells were treated with different concentrations of BPA for 12, 24, and 48 h, respectively, and the 20% inhibitory concentration (IC20) was measured using a CCK-8 assay. The expression levels of piRNAs before and after BPA treatment were determined by reverse transcription-quantitative PCR. Target genes regulated by BPA and associated with prostate cancer were screened in the Comparative Toxicogenomics Database (CTD). Dual-luciferase reporter gene assay was performed to verify the relationship between piRNA and target genes, and the expression change of the piRNA target gene was detected by Western blotting. Cell migration and invasion assays were used to determine the effects of piRNA on the malignant phenotype of prostate cancer cells. Results: After treatment of PC-3 cells with 160 µmol/L BPA, the expression of piR-sno48 was most significantly increased (P<0.05). Transfection of piR-sno48 antagomir resulted in decreased expression of endogenous piR-sno48 and a significant increase in the expression of its target gene GSTP1 (P<0.05). However, the expression of GSTP1 did not change significantly in BPA-treated PC-3 cells after transfection with piR-sno48 antagomir (P>0.05). The dual-luciferase reporter gene confirmed that piR-sno48 inhibited the expression of GSTP1 by forming an inversely complementary sequence with the 3'-UTR of GSTP1. The Transwell assay results showed that treatment with BPA significantly increased the invasion and migration ability of prostate cancer cells (P<0.01), whereas piR-sno48 antagonists significantly inhibited the effects above (P<0.01). Conclusion: BPA promotes the invasion and migration of prostate cancer cells by upregulating the expression of piR-sno48 and suppressing the expression of GSTP1. Interfering with the expression of endogenous piR-sno48 may inhibit the malignant phenotype of prostate cancer cells caused by BPA.

[Characteristics and regulatory mechanisms of lipid metabolism remodeling after malignant transformation of glioma-associated macrophages].

Objective: To observe the lipid metabolism characteristics of tumor-associated macrophages (TAM) after malignant transformation in the glioma micro-environment, and analyze the biological phenotype changes and regulatory mechanisms after inhibiting the lipid metabolism remodeling. Methods: Twelve male Balb/c mice of 6-8 weeks were used in the study. Macrophages (Mφ) were derived from mouse bone marrow, and malignantly transformed macrophages (tMφ1 and tMφ2) were cloned from the model of glioma stem cell (GSC) through interaction with Mφ in vivo and in vitro. Intracellular lipid droplet formation and cellular cholesterol content were measured respectively in Mφ, tMφ1 and tMφ2. qRT-PCR was performed to detect the genes expression level related with lipid metabolism, including sterol regulatory element binding protein (SREBP), fatty acid synthase (FASN), and 3-hydroxy-3-methylglutarate monoacyl coenzyme A reductase (HMG-CoA). Simvastatin (SIM) was used to analyze the proliferation, immigration and invasiveness ability in tMφ1 and tMφ2 after inhibition of the lipid metabolism. Differential expression profiles of miRNAs after SIM treatment were constructed in t-Mφ1 and bio-informatics analysis was screened and verified for miR449a and its target gene sorting micro-tubule connectin 17 (SNX17) associated with lipid metabolism remodeling. The effect on SNX17 by up-regulated miR-449a were analyzed by qRT-PCR and Western blot, meanwhile, the biological phenotype and cholesterol content were observed after up-regulation of miR449a. Low-density lipoprotein receptor (LDLR) protein levels after SNX17 knockdown and intracellular cholesterol content after LDLR knockdown were detected respectively. Results: The numbers of intracellular lipid droplet formation in tMφ1 and tMφ2 were more than that in Mφ (P<0.001). Likewise, the relative contents of cholesterol (3.89±0.68 and 3.56±0.53), SREBP (4.78±0.60 and 2.84±0.41), FASN (4.65±0.70 and 3.01±0.45), and HMG-CoA (5.74±0.55 and 2.97±0.34) were significantly higher in tMφ1 and tMφ2 than those of Mφ (1.01 wel, 1.02 wel and 0.99 wel, respectively) (all P<0.001). The proliferation rates of tMφ1 and tMφ2 decreased from (47.06±5.88) % and (45.29±5.64)% to (23.53±4.70)% and (18.74±5.76)%, respectively after treatment with SIM (both P<0.05). The numbers of migrated cells decreased from 1 025±138 and 350±47 to 205±63 and 99±25, respectively (both P<0.001). And the numbers of invasiveness cells decreased from 919±45 and 527±34 to 220±23 and 114±21, respectively (both P<0.001). While the relative intracellular cholesterol content decreased to 0.52±0.08 and 0.58±0.07 (both P<0.05), respectively. MiR-449a was screened from tMφ1 by SIM, and the target gene was analyzed and verified to be SNX17. SNX17 expression was down-regulated, and the proliferation rate, the number of migration and invasiveness was significantly decreased after miR-449a over-expression (all P<0.05). Low-density lipoprotein receptor (LDLR) expression was down-regulated after knock-down of SNX17, while the cholesterol content was decreased after knock-down of LDLR in tMφ1 and tMφ2 (all P<0.05). Conclusions: Malignantly transformed TAMs undergo lipid metabolism remodeling characterized with enhanced lipid metabolism. MiR-449a regulates the LDLR by targeting SNX17, thereby affecting the lipid metabolism of malignantly transformed macrophages, and subsequently inhibiting its proliferation, migration, and invasion ability. Precise intervention with miR-449a/SNX17/LDLR axis could provide an experimental basis for reversing its tumor-promoting micro-environment remodeled by GSC through metabolic intervention.

[Research progress of new generation intraocular lens calculation formulas].

With the development of the technology of phacoemulsification and intraocular lens (IOL) implantation, the calculation formula of IOL power has been optimized. The existing formulas are still not accurate enough for some special cases, such as patients with short or long axial length and previous corneal refractive surgery, because the calculation of IOL power is a nonlinear function, and these mathematical formulas based on an optical model can not show the nonlinear relationship well. Therefore, new IOL calculation formulas developed by using an artificial intelligence nonlinear mathematical model have come into being. This review demonstrates the development and application of new IOL calculation formulas, in order to provide guidance for improving the visual quality and clinical application of each calculation method in cataract patients.

[Analysis of risk factors for gestational diabetes mellitus in elderly multipara women in the next pregnancy].

Objective: To investigate the risk factors for gestational diabetes mellitus (GDM) in elderly multipara women in the next pregnancy. Methods: A total of 219 elderly multipara women with 2 consecutive delivery records in Tianjin Binhai New Area Tanggu Obstetrics and Gynecology Hospital from January 2018 to May 2019 were included. Among them, 141 had normal glucose tolerance (NGT) and 78 of them had GDM. The clinical data of the previous and current pregnancy were collected to analyze the risk factors of GDM in elderly multipara women. Results: The average ages of 219 elderly women in previous pregnancy and this pregnancy were (31.9±2.2) and (36.7±1.5) years old, and the prevalence of GDM was 35.62% (78 cases). Compared to NGT group, GDM patients had higher fasting blood glucose(previous (5.51±1.43) vs (4.63±0.62) mmol/L; current (5.26±0.63) vs (4.59±0.30) mmol/L, 1 h blood glucose(previous (11.74±2.36) vs (9.50±1.82) mmol/L; current (11.03±2.03) vs (9.51±1.14) mmol/L) in 75 g oral glucose tolerance test (OGTT) in both previous and current pregnancy. The rates of cesarean section, in both previous and current pregnancy were higher in GDM group (previous 34.6% vs 4.3%; current 52.6% vs 22.0%). Furthermore, prenatal weight and body mass index (BMI) of the previous pregnancy, pre-pregnancy weight and BMI, and prenatal BMI of this pregnancy were also higher in GDM group, and the differences were all statistically significant (all P<0.05). Logistic multivariate regression analysis indicated cesarean section history (OR=10.80, 95%CI: (4.09-28.54)), GDM history of previous pregnancy (OR=10.64, 95%CI: (4.02-28.20)), 75 g OGTT fasting blood glucose≥ 4.86 mmol/L (OR=2.70, 95%CI: (1.27-5.70)), 1 h blood glucose after glucose administration ≥ 8.45 mmol/L (OR=1.78, 95%CI: (1.37-2.31)) were risk factors for GDM in elderly multipara women of this pregnancy. Conclusion: The risk of GDM in elderly multipara women with a history of cesarean section and GDM increases significantly. Results of OGTT in previous pregnancy also has predictive value.

[Role of exosomal miRNA in tumor progression and its possible mechanism].

Exosome is a kind of biological nano-vesicle with a diameter of about 30-100 nm and is synthesized, secreted, and released in almost all types of body cells. Recent studies have found that exosomes contain a variety of functional proteins, including mRNA and microRNAs (miRNAs) and so on, which play an important role in the process of material transfer and information exchange in cells. In addition, the related-exosomal microRNAs secreted by tumor cells play an important role in regulating theoccurrence, development, invasion and metastasis of tumors and other biological processes.. Therefore, the study of tumor-related exosomal microRNAs will help us to explore the mechanism of tumor progression from the perspective of tumor gene regulation and new tumor markers, which will contribute to the early diagnosis, treatment, disease monitoring and prognosis evaluation.

The effect of androgen on wool follicles and keratin production in Hetian sheep / Efeito do androgênio na produção de folículos e queratina em ovinos Hetianos

Abstract To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.

Resumo Investigar a concentração ideal de andrógenos em cultura de folículos pilosos de carneiro Hetiano e detectar os efeitos da concentração de andrógenos na proliferação e apoptose de células foliculares, por meio de imunofluorescência e de determinação quantitativa, em tempo real, da fluorescência dos níveis de expressão gênica de proteína associada à queratina. Folículos pilosos foram isolados por microdissecção, e folículos de lã e pedaços de pele foram cultivados em várias concentrações de di-hidrotestosterona (DHT) em meio de cultura. Em seguida, medições diárias de crescimento longitudinal dos folículos capilares foram obtidas usando um micrômetro microscópico. Folículos de lã cultivados de Hetianos foram corados pelo método SACPIC para revelar a estrutura do folículo piloso, enquanto fatias de pele de carneiro foram usadas para observar a proliferação celular por imunocoloração e apoptose celular por meio do método TUNEL. Em âmbito da biologia molecular, a expressão gênica da proteína associada à queratina (Kap) foi estudada usando folículos capilares cultivados por vários dias, in vitro. Os efeitos das concentrações de andrógenos no crescimento e desenvolvimento dos folículos de lã de Hetianos foram estudados experimentalmente. Ensaios de proliferação de EdU revelaram que o andrógeno promoveu a proliferação celular dentro das papilas dérmicas do folículo piloso. A detecção de apoptose por TUNEL demonstrou que o tratamento com andrógeno poderia atrasar a apoptose celular. Os resultados da reação em cadeia da polimerase transcrição reversa quantitativa (qPCR) demonstraram que os padrões de expressão gênica da proteína de enxofre Kap1.1, KIF1.2, Kap2.12 e Kap4.2 foram significativamente maiores no grupo de ovinos Hetianos de montanha. Uma concentração de androgênio de 100 nM pode promover o crescimento de células foliculares de lã de Hetianos in vitro, resultando na superexpressão de alguns genes da família Kap.

Investigating the Potential Mechanism of Pydiflumetofen Resistance in Sclerotinia sclerotiorum.

The necrotrophic pathogen Sclerotinia sclerotiorum is one of the most damaging and economically important plant pathogens. Pydiflumetofen, which was developed by Syngenta Crop Protection, has already been registered in China for the management of Sclerotinia stem rot, which was caused by S. sclerotiorum in oilseed rape. In an attempt to preempt and forestall the development of resistance to this useful fungicide, the current study was initiated to investigate the potential mechanism of resistance in laboratory mutants. Five pydiflumetofen-resistant S. sclerotiorum mutants were successfully generated by repeated exposure to the fungicide under laboratory conditions. Although the mutants had greatly reduced sensitivity to pydiflumetofen, they were also found to have significantly (P < 0.05) reduced fitness, exhibiting reduced mycelial growth and sclerotia formation on potato dextrose agar medium. However, three of the four mutants had significantly (P < 0.05) increased pathogenicity on detached soybean leaves compared with their respective parental isolates, indicating a moderate to high level of fungicide resistance risk according to the criteria of the Fungicide Resistance Action Committee. Sequence analysis of four succinate dehydrogenase (Sdh) target genes identified several nucleotide changes in the sequences of the pydiflumetofen-resistant mutants, most of which were synonymous and caused no changes to the predicted amino acid sequences. However, all of the pydiflumetofen-resistant mutants had two amino acid point mutations (A11V and V162A) in their predicted SsSdhB sequence. No similar changes were found in the SsSdhA, SsSdhC, and SsSdhD genes of any of the mutants tested. In addition, there was a positive cross-resistance between pydiflumetofen and boscalid, and no cross-resistance between pydiflumetofen and other commonly used fungicides, including tebuconazole, fludioxonil, cyprodinil, dimethachlone, prochloraz, pyraclostrobin, fluazinam, procymidone, and carbendazim. These results indicate that pydiflumetofen has great potential as an alternative fungicide for the control of S. sclerotiorum, especially where resistance to other fungicides has already emerged. Mixing or alternate application with fludioxonil, prochloraz, and fluazinam could be used to limit the risk of resistance to pydiflumetofen.

[A case of hand, foot and mouth disease in adults who first diagnosed with conjunctivitis].

A 34-year-old female presented to the doctor with chief complaint of redness in both eyes for 1 week. The slit lamp examination showed slight eyelid swelling, water-like discharge in the conjunctival sac of the lower eyelid, conjunctival follicles, gray-white pseudomembrane and small lamellar subconjunctival hemorrhage on the lower palpebral conjunctiva. Physical examination showed two erythematous macules(size 5 mm×6 mm) on the left palm. One erythematous macule(size 4 mm×5 mm) was seen on the right palm. Multiple erosions were located in the skin of upper and lower lips, oral cavity and the gingival mucosa. A diagnosis of hand, foot and mouth disease was made following by dermatological consultation. After topical antiviral and steroids treatment, the disease quickly relieved.(Chin J Ophthalmol, 2021, 57: 139-141).

The effect of androgen on wool follicles and keratin production in Hetian sheep.

To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.

[Observation of cosmetic effect of corneal interlamellar staining in patients with corneal leucoma].

Objective: To observe the clinical effect of corneal staining in patients with corneal leucoma. Methods: Restrospective case series study.Thirty eyes of 30 patients (17 males, 13 females) with corneal leucoma who underwent corneal interlamellar staining at Beijing Aier-Intech Eye Hospital from October 2014 to July 2018 were included. The mean age was 31.50±15.66 years. Postoperative follow-up was more than 1 year. All the patients underwent appearance examination, anterior segment examination with a slit lamp, B-scan examination and anterior segment OCT examination preoperatively and postoperatively. The effect of corneal staining, patient satisfaction and the incidence of complications were investigated. Results: The corneal epithelium of all the patients healed completely within 1-3 days after operation. The mean follow-up period was 30.68±18.02 months (range, 12.37-58.10 months). During the postoperative follow-up period, no staining permeation or inflammatory reaction in the anterior chamber occurred. The corneal color and appearance were well maintained in 18 patients (72.0%). Seven patients (28.0%) showed mild corneal color-fading. One patient was treated with enucleation and orbital hydroxyapatite implantation because of eye atrophy and corneal banding degeneration caused by the primary disease at 4 years after operation. The surgeon was satisfied with the improvement of the appearance of all cases; 22 patients (88.0%) were satisfied with the postoperative appearance. No significant complications were observed in all the cases. Conclusions: Corneal interlamellar staining is one of best choices for the treatment of corneal leucoma. It has advantages of quick postoperative recovery, long-standing color staining and good cosmetic effect. The operation is simple and easy to carry out and there is no obvious damage to eye tissues. (Chin J Ophthalmol, 2020, 56:465-472).

Locomotor transition: how squid jet from water to air.

The amazing multi-modal locomotion of flying squid helps to achieve fast-speed migration and predator-escape behavior. Observation of flying squid has been rarely reported in recent years, since it is challenging to clearly record the flying squid's aquatic-aerial locomotion in a marine environment. The existing reports of squid-flying events are rare and merely record the in-air motion. Therefore, the water-air locomotor transition of flying squid is still unknown. This paper proposes the idea of using CFD to simulate the process of the flying squid (Sthenoteuthis oualaniensis (S. oualaniensis)) launching from water into air. The results for the first time reveal the flow field information of squid in launching phase and show the kinematic parameters of flying squid in quantification. Both a trailing jet and pinch-off vortex rings are formed to generate launching thrust, and the formation number L ω /D ω is 5.22, demonstrating that the jet strategy is to produce greater time-averaged thrust rather than higher propulsion efficiency. The results also indicate that the maximum flying speed negatively correlates with the launch angle, indicating that a lower launch angle could result in a larger flying speed for the flying squid to escape. These findings explore the multi-modal locomotion of flying squid from a new perspective, helping to explain the trade-off strategy of water-to-air transition, and further enhance the performance of aquatic-aerial vehicles.

Piezoelectric modulation of broadband photoresponse of flexible tellurium nanomesh photodetectors.

Flexible photodetector shows great potential applications in intelligent wearable devices, health monitoring, and biological sensing. In this work, single crystal ß-tellurium nanowires were grown on flexible muscovite by molecular beam epitaxy, constructing high-density ordered nanomesh structure. The prepared photodetectors based on tellurium nanomesh exhibit excellent mechanical flexibility, fast response in a broad range from ultraviolet to near-infrared, and good photosensitivity. We found that the flexible photodetectors with Shottky contact drastically suppressed dark current, while the response speed was lowered in comparison to the devices with ohmic contact, as holes would take a long time to tunnel through the Shottky barrier between metal and p-type Te. Moreover, the photoresponse of flexible Shottky photodetectors can be modulated by piezoelectricity of tellurium, and pronounced photocurrent increase after bending many times. Under external stress, polarization charges could tune Shottky barrier height of the metal/tellurium, resulting in variation of photocurrent. This research not only explores the broadband photoresponse and piezoelectric effect of tellurium nanomesh, but also promotes the integration and development of broadband flexible optoelectronic devices.

[The mechanism and role of autologous peritoneum transplantation to enterocystoplasty in porcine model].

Objective: To explore the pathological mechanism and role of autologous peritoneum transplantation onenterocystoplasty in porcine model. Methods: Randomly, four experimental female swine were chosen, and each of their body weight was 28-33 kg. By intravenous anesthesia, the transplantation of autologous peritoneum for bladder reconstruction operation was carried out (to transplant the peritoneum onto an ileum segment of which the mucosa and submucosa had been removed. These flaps were used to mend and augment the bladder by suturing the edge of detective bladders). After removal of ureteral catheters and balloon catheter at day 5 respectively, their voiding behaviors were monitored, and respectively 2 swine were euthanized at week 3/6 for routine pathology, immunohistochemistry, and electron microscopic examinations. Results: Four swine underwent reconstruction, but none died for complications such as peritonitis, ileus and urinary fistula. The length of ileum was 35 cm, and the area of peritoneum was 15 cm×10 cm. Voiding behaviorsof the swine were normal, with clear urine after removal of their catheters. At autopsy, reconstructed bladders were healthy. Pathological examination showed the neobladder had been covered by continuous urothelium while the peritoneum disappeared without ileal mucosa regrowth or residual. Scanning electron microscope showed that the transitional cells of neobladder of swine were complete and orderly, and urothelium around suture border was continuous withoutmalposition. Conclusions: In porcine model, autologous peritoneal transplantation witnessed no immune rejection response for itself, which was replaced by crawling transitional epithelium around anastomosis, rather than by metaplasia.

[An animal model of bladder reconstruction by autologous peritoneum transplantation].

Objective: To establish experimental porcine model of reconstruction the neobladder by ileal seromuscular with transplantation of autologous peritoneum. Methods: This was an animal experiment carried out from January to April 2018 at animal center of Guizhou Medical University. Randomly 6 experimental female porcines were chosen, and their body weight was 28 to 33 kg. By intravenous anesthesia, the transplantation of autologous peritoneum for bladder reconstruction operation was carried out by transplanting the peritoneum onto an ileum segment which mucosa and submucosa had been removed. These flaps were used to mend and reconstruct the neobladder by suturing with edge of the detective bladder. After removal of ureteral catheters and balloon catheter at day 5 and day 7 respectively, voiding behavior was monitored, and animals were euthanized at week 12 for routine pathology, immunohistochemistry, and electron microscopic examinations. Results: Six porcines underwent reconstruction, but no one lost to complications such as peritonitis, ileus and urinary fistula. Voiding behavior was normal, and urine was clear in all animals after removal of catheters. At autopsy, reconstructed bladders were healthy. Pathological examination showed the part of reconstruction had been covered by continuous urothelium while the peritoneum disappeared and showed no ileal mucosa regrowth and residual. Scanning electron microscope showed that the transitional cells of neobladder were complete and orderly, and urothelium around suture border was continuous and no malposition. Conclusion: In this experimental porcine model, reconstruction bladder by autologous peritoneum and ileal seromuscular flaps is an ideal approach.

[Study on correlation between serum 25-hydroxyvitamin D3 level and esophageal variceal bleeding in cirrhotic patients].

Objective: To explore the correlation between serum 25-hydroxyvitamin D3 (25[OH]D(3)) levels and esophageal variceal bleeding (EVB) in cirrhotic patients. Methods: Eighty-three cases with liver cirrhosis hospitalized from November 2016 to January 2017 were collected. The patients were divided into bleeding group (51 cases) and non-bleeding group (32 cases) depending on the presence or absence of bleeding under gastroscopy. Serological tests were performed on both groups, including hemoglobin (Hb), albumin (ALB), alkaline phosphatase (ALP),γ-glutamyltransferase (GGT), interleukin-6 (IL-6), and 25-hydroxyvitamin D3 (25[OH]D(3)). Both groups were analyzed by univariate analysis. The differences between both groups were compared by t-test, after normality test. The other variables were compared by Mann-Whitney U test. The correlation between the relevant variables and EVB were analyzed by Spearman's rank correlation and a multivariate analysis. Cases with primary biliary cirrhosis were relatively low in number (four cases in bleeding group, accounting for 8%, 10 cases in non-bleeding group, accounting for 31%). The effects of ALP and GGT on serum 25(OH)D(3) level were analyzed by stratified analysis. Moreover, ALP and GGT levels were divided into two and three groups: < 140 U/L and >140 U/L and < 30 U/L, > 30 U/L, and ~≤60 U/L. Results: Bleeding group had low levels of hemoglobin (t= -2.827,P= 0.005), alkaline phosphatase (t= -3.097,P= 0.002), gamma-glutamyltransferase (t= -2.292,P= 0.022), and 25(OH)D(3) (t= -3.134,P= 0.002) than non-bleeding group. Both groups (P> 0.05) had similar levels of albumin, interleukin-6, AAR, and FIB-4. Logistic regression analysis showed that 25(OH)D(3), alkaline phosphatase and hemoglobin were independent risk factors for EVB. Spearman's correlation coefficient analysis showed that 25(OH)D(3)was significantly positively and negatively correlated with interleukin-6 (r= 0.306,P= 0.005) and albumin (r= -0.327,P= 0.003). Stratified analysis showed that serum 25(OH)D(3) level was lower in ALP≤140U/L group and the bleeding group, and the difference was statistically significant than non-bleeding group (P= 0.007), while the serum level of 25(OH)D(3)was decreased in both groups for alkaline phosphatase > 140 U/L group, and the difference was not statistically significant (P= 0.051). Furthermore, in the GGT > 60 U/L group, the serum level of 25(OH)D(3)was significantly lower in the bleeding group, and the difference was statistically significant in non-bleeding group (P= 0.003), while the difference between the two groups was not statistically significant (P> 0.05) in GGT≤30 U/ L, > 30 U/L, and ~≤60 U/L group. Conclusion: Serum 25(OH)D(3)level was significantly lower in EVB cirrhotic patients, and it was an independent risk factor for EVB. Serum 25(OH)D(3) low levels was more apparent with ALP normalization or GGT level > 60 U/L.

[Establishment and evaluation of a triple-color human papillomavirus pseudovirion neutralization assay].

Objective: To establish a triple-color pseudovirion-based neutralization assay (PBNA) and evaluate its capability of detecting immunogenicity of the sera generated by the immunization of HPV 9-valent vaccine. Methods: HPV pseudovirus (PsVs) 6/11/16/18/31/33/45/52/58 with the encapsidated fluorescence expressing red fluorescent plasmid N31-MCHREEY, green fluorescent N31-EGFP or blue fluorescent N31-mTagBFP were generated. The concentration of HPV PsVs and the infection titers of HPV PsVs were detected by double-antibody sandwich ELISA and TCID(50), respectively. The single- and triple color HPV 16/33/45 PsVs were used to detect the neutralization titers of mice sera immunized with HPV 9-valent vaccine and confirmed the accuracy and specificity of the triple-color PBNAs. Then, the single- and triple color HPV 6/11/18/31/33/45/52/58 PsVs were employed to detect the neutralization titers of cynomolgus macaques sera immunized with HPV 9-valent vaccine and determined whether the triple-color PBNAs could be applied to evaluate the immunogenicity of the sera generated by the immunization of HPV9-valent vaccine. Results: The concentration of HPV16 PsVs encapsulating green, red or blue fluorescent plasmid was 5.0 to 6.0 µg/ml and HPV6/11/18/31/33/45/52/59 triple-color HPV PsVs was about 1.0 to 3.0 µg/ml. 9 types HPV PsVs containing EGFP, Mcherry or mTagBFP reporter plasmid were obtained and the concentration can meet the need of neutralization detection. 9 types single-color fluorescent HPV PsVs had similar infectivity against 293FT cells with the infection titer values between 1×10(4) and 1×10(5). The results of PBNAs showed that there was no significant difference in the anti-HPV neutralization titers of mice sera induced by HPV 9-valent vaccine between single-color and triple-color HPV16/33/45 PsVs (P>0.05). Similarly, there was also no significant difference in the anti-HPV neutralization titers of cynomolgus macaques sera induced by HPV 9-valent vaccine between single-color and triple-color HPV6/11/18/31/33/45/52/58 PsVs (P>0.05). Conclusion: We successfully established the triple-color PBNAs and verified the accuracy and specificity of triple-color PBNAs consistent with single-color PBNAs. The triple-color PBNAs can be applied to evaluate the immunogenicity of HPV 9-valent vaccine's immune serum.

High expression of long non-coding RNA LOC730101 correlates with distant metastasis and exhibits a poor prognosis in patients with osteosarcoma.

OBJECTIVE: Expression of long non-coding RNA LOC730101 (LOC730101) has been closely linked to the carcinogenesis of osteosarcoma and has been shown to function as a tumor promoter. However, the clinical significance of LOC730101 remains unknown. Our present study aimed to investigate the feasibility of LOC730101 as a novel prognostic biomarker for osteosarcoma. PATIENTS AND METHODS: Quantitative Real-time RT-PCR (qRT-PCR) was performed to examine LOC730101 expression in osteosarcoma tissues and adjacent non-cancerous tissues. The correlation between LOC730101 expression and clinicopathological features and the overall survival rate was determined. Cox regression analyses were performed to explore whether LOC730101 was an independent predictor of survival for osteosarcoma patients. RESULTS: We found that LOC730101 was significantly upregulated in human osteosarcoma tissues compared with corresponding adjacent normal tissues (p < 0.01). Elevated LOC730101 expression was correlated with advanced clinical stage (p = 0.001) and distant metastasis (p = 0.005). Furthermore, survival assay revealed that osteosarcoma patients in the high LOC730101 expression group had significantly shorter 5-year overall survival time than those in the low LOC730101 expression group (p = 0.0002). In addition, multivariate analysis showed that LOC730101 maintained an independent prognostic influence on overall survival (p = 0.006). CONCLUSIONS: Our present study, for the first time, revealed that LOC730101 may serve as a poor prognostic indicator in patients with osteosarcoma.

Development and validation of a new cognitive screening test: The Hong Kong Brief Cognitive Test (HKBC).

OBJECTIVES: To develop and examine the validity of a new brief cognitive test with less educational bias for screening cognitive impairment. METHODS: A new cognitive test, Hong Kong Brief Cognitive Test (HKBC), was developed based on review of the literature, as well as the views of an expert panel. Three groups of subjects aged 65 or above were recruited after written consent: normal older people recruited in elderly centres, people with mild NCD (neurocognitive disorder), and people with major NCD. The brief cognitive test, Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment Scale (MoCA), were administered to the subjects. The performance of HKBC in differentiating subjects with major NCD, mild NCD, and normal older people were compared with the clinical diagnosis, as well as the MMSE and MoCA scores. RESULTS: In total, 359 subjects were recruited, with 99 normal controls, 132 subjects with major NCD, and 128 with mild NCD. The mean MMSE, MoCA, and HKBC scores showed significant differences among the 3 groups of subjects. In the receiving operating characteristic curve analysis of the HKBC in differentiating normal subjects from those with cognitive impairment (mild NCD + major NCD), the area under the curve was 0.955 with an optimal cut-off score of 21/22. The performances of MMSE and MoCA in differentiating normal from cognitively impaired subjects are slightly inferior to the HKBC. CONCLUSIONS: The HKBC is a brief instrument useful for screening cognitive impairment in older adults and is also useful in populations with low educational level.

A wind tunnel study of effects of twisted wind flows on the pedestrian-level wind field in an urban environment.

The influence of twisted wind flows on the pedestrian-level wind (PLW) field of an urban area was evaluated by testing a typical urban site (Tsuen Wan, Hong Kong) in a boundary layer wind tunnel. Four twisted wind profiles with different magnitudes and directions of yaw angles were employed to investigate variations in wind speed with the properties of the twisted wind flows at the pedestrian level. An additional conventional wind profile with similar wind speeds and turbulence intensities to the twisted winds but with zero yaw angles was simulated for comparisons. The mean wind speeds at 77 locations including the perimeter, roadsides, and groups of high-rise buildings were analysed for the conventional and the four twisted wind flows. The comparisons show a tendency of twisted winds to generate higher wind speeds at the pedestrian level than the conventional wind profile. The wind speeds of the twisted winds have a strong dependence on the magnitude and direction of the yaw angles, particularly at locations where the densities of buildings in the neighbourhood are low and hence local wind circulations are significantly modified by the twisted winds.