Acupuncture and Chinese herbal medicine for menopausal mood disorder: a randomized controlled trial.

OBJECTIVE: This study aimed to analyze the effectiveness of acupuncture combined with Chinese herbal medicine (CHM) on mood disorder symptoms for menopausal women. METHODS: A total of 95 qualified Chinese participants were randomly assigned to one of three groups: 31 in the acupuncture combined with CHM group (combined group), 32 in the acupuncture combined with CHM placebo group (acupuncture group) and 32 in the CHM combined with sham acupuncture group (CHM group). The patients were treated for 8 weeks and followed up for 4 weeks. The data were collected using the Greene Climacteric Scale (GCS), self-rating depression scale (SDS), self-rating anxiety scale (SAS) and safety index. RESULTS: The three groups each showed significant decreases in the GCS, SDS and SAS after treatment (p < 0.05). Furthermore, the effect on the GCS total score and the anxiety domain lasted until the follow-up period in the combined group (p < 0.05). Within the three groups, there was no difference in GCS and SAS between the three groups after treatment (p > 0.05). However, the combined group showed significant improvement in the SDS, compared with both the acupuncture group and the CHM group at 8 weeks and 12 weeks (p < 0.05). No obvious abnormal cases were found in any of the safety indexes. CONCLUSIONS: The results suggest that either acupuncture, or CHM or combined therapy offer safe improvement of mood disorder symptoms for menopausal women. However, the combination therapy was associated with more stable effects in the follow-up period and a superior effect on improving depression symptoms.

[Association of urinary sodium excretion with fluid overload in patients with chronic kidney disease].

Objective: To investigate the relationship between urinary sodium excretion and fluid overload (FO) in non-dialysis patients with chronic kidney disease (CKD). Methods: Patients with CKD stage 1-4 who underwent bioelectrical impedance (BIA) in the Department of Nephrology, Jiangsu Province Hospital from December 2019 to January 2021 were recruited. All enrolled patients were categorized into two groups according to whether or not they develop FO. Further, clinical parameters were compared between the two groups. Spearman correlation analysis was used to investigate the association between over hydration/extracellular water (OH/ECW) and clinical characteristics. Multivariate logistic regression analysis was performed to evaluate the relationship between urinary sodium excretion and FO (FO was defined as OH/ECW≥7%). Results: A total of 385 patients with CKD stage 1-4 were finally included in the study, with a mean age of (46±15) years. There were 216 male cases (56.1%), and 150 cases (39.0%) existed FO. Spearman correlation analysis indicated that OH/ECW positively correlated with urinary sodium excretion (r=0.147, P=0.004), urinary protein excretion (r=0.555, P<0.001) and systolic blood pressure (r=0.241, P<0.001), but inversely related to estimated glomerular filtration rate (eGFR) (r=-0.111, P=0.030) and serum albumin (r=-0.659, P<0.001). After adjusting for confounding factors including age, systolic blood pressure, diabetes, urinary protein excretion, serum albumin, serum sodium, serum chlorine, urinary calcium excretion, urinary phosphorus excretion and use of diuretics, multivariate logistic regression analysis demonstrated that higher level of urinary sodium excretion was associated with increased risk of FO in patients with CKD (OR=1.005, 95%CI: 1.000-1.011, P=0.048). Conclusion: High urinary sodium excretion is independently associated with fluid FO in non-dialysis patients with CKD.

Second- and third-trimester serum levels of growth-differentiation factor-15 in prediction of pre-eclampsia.

OBJECTIVE: Pre-eclampsia (PE) is a significant contributor to adverse maternal and perinatal outcome; however, accurate prediction and early diagnosis of this condition remain a challenge. The aim of this study was to compare serum levels of growth-differentiation factor-15 (GDF-15) at three different gestational ages between asymptomatic women who subsequently developed preterm or term PE and healthy controls. METHODS: This was a case-control study drawn from a prospective observational study on adverse pregnancy outcomes in women attending for their routine second- and third-trimester hospital visits. Serum GDF-15 was determined in 300 samples using a commercial GDF-15 enzyme-linked immunosorbent assay: 120 samples at 19-24 weeks of gestation, 120 samples at 30-34 weeks and 60 samples at 35-37 weeks. Multiple linear regression was applied to logarithmically transformed GDF-15 control values to evaluate the influence of gestational age at blood sampling and maternal characteristics on GDF-15 results. GDF-15 multiples of the normal median (MoM) values, adjusted for gestational age and maternal characteristics, were compared between pregnancies that subsequently developed preterm or term PE and healthy controls. RESULTS: Values of GDF-15 increased with gestational age. There were no significant differences in GDF-15 MoM values between cases of preterm or term PE and normotensive pregnancies at 19-24 or 35-37 weeks of gestation. At 30-34 weeks, GDF-15 MoM values were significantly increased in cases of preterm PE, but not in those who later developed term PE. Elevated GDF-15 MoM values were associated significantly with a shorter interval between sampling at 30-34 weeks and delivery with PE (P = 0.005). CONCLUSION: Serum GDF-15 levels at 19-24 or 35-37 weeks of gestation are not predictive of preterm or term PE. At 30-34 weeks, GDF-15 levels are higher in women who subsequently develop preterm PE; however, this difference is small and GDF-15 is unlikely to be useful in clinical practice when used in isolation. Copyright © 2020 ISUOG. Published by John Wiley & Sons Ltd.

[Effects of Prrx2 gene silencing on the proliferation of breast cancer and its molecular mechanisms].

Objective: The aim of this study was to investigate the effects of silencing Paired related homoeobox 2 (Prrx2) expression on the proliferation of breast cancer and its molecular mechanisms. Methods: Short hairpin RNA knockdown of Prrx2 was used to examine cellular effects of Prrx2. The level of Prrx2 was verified by Western blot. MTT assay was used to analyze the proliferation of breast cancer cells in vitro. To investigate the effect of Prrx2 depletion on tumor growth in vivo, a nude mouse xenograft model was performed. Results: The expression of Prrx2 decreased 91.2% in MDA-MB-231 cells and 88.7% in MCF-7 cells after transfection with interfering vectors (P<0.05). MTT assay showed that the proliferation of cells in silenced Prrx2 expression group was significantly inhibited compared with the control group (P<0.05). Nude mice transplanted tumors showed that the growth of transplanted tumors was slow after silencing Prrx2 expression, and the weight of the tumors of silenced Prrx2 expression group were smaller than those of the control group ((160.2±26.3)mg vs (365.4±19.7)mg, P<0.05). Western blot showed that silencing Prrx2 expression inhibited the expression of ß-catenin in breast cancer cell nucleus and down-regulated the activity of Wnt/ß-catenin signaling pathway. Conclusions: Silencing Prrx2 expression can effectively inhibit the proliferation and growth of breast cancer, suggesting that Prrx2 may become a new target for the treatment of breast cancer.

Molecular cloning, expression and functional characterization of tumor necrosis factor (TNF) receptor-associated factor (TRAF)-interacting protein (TRIP) in grass carp, Ctenopharyngodon idella.

TRIP (Tumor Necrosis Factor (TNF) Receptor-Associated Factor (TRAF)-Interacting Protein), a member of the TNF superfamily, plays a crucial role in the modulation of inflammation in vertebrates. However, no information about TRIP is available in teleosts. In this study, the full-length cDNA of TRIP, containing a 5'UTR of 112 bp, an ORF of 1359 bp, and a 3'UTR of 29 bp before the poly (A) tail, was cloned from grass carp, Ctenopharyngodon idella. The TRIP gene encoded a protein of 452 amino acids with an estimated molecular mass of 51.06 KD and a predicted theoretical isoelectric point (pI) of 9.11. Quantitative real-time PCR analysis revealed that TRIP mRNA was expressed in all the tissues examined in grass carp, with the highest expression in the kidney, followed by the intestine and thymus. However, lower levels of expression were also detected in fat, spleen, liver, gonad and heart. Subcellular localization and two-hybrid analysis revealed that TRIP was located in the nucleus and that it interacted with TRAF1 and TRAF2 in HEK293T cells. Furthermore, similar to TNF-α, IL-10 and TRIP mRNA expression was upregulated in the spleen of fish fed high-fat or high-carbohydrate diets, suggesting that TRIP might be associated with the response to excessive energy intake. The mRNA relative expression of TRIP was significantly reduced (P < 0.05) after hepatocyte of C. idella was treated with 2 µg/mL lipopolysaccharide (LPS) for 4 h, while the expression levels of inflammatory cytokines TNF-α and IL-10 were significantly increased (P < 0.05). Taken together, these results indicate that TRIP might play important roles in immune defense and has the potential to be used as a anti-inflammation target in grass carp.

Molecular characterization and expression analysis of the ß-actin gene from the ridgetail white prawn Exopalaemon carinicauda.

Actin is a highly conserved protein that is found in all eukaryotic cells, and has been widely used as an internal control gene in gene expression studies. In this study, we cloned an actin gene (named Ecß-actin) from Exopalaemon carinicauda and determined its expression levels. The full-length cDNA of Ecß-actin was 1335 bp long, comprising a 1131-bp ORF encoding 376 amino acids, a 65-bp 5'-UTR, and a 139-bp 3'-UTR with a poly(A) tail. The A + T content was approximately 79% in the 3'-UTR of the Ecß-actin mRNA. The 3'-UTR contained two repeats of the AUUUA motif. The putative protein Ecß-actin showed high identity (97-99%) with other actins from various species. Phylogenetic analysis revealed that Ecß-actin belongs to Crustacea, although it formed a singleton sub-branch that was located a short distance from crabs and other shrimp species. Ecß- actin expression was detected in the hepatopancreas, ovary, muscle, gill, stomach, and hemocytes, and was strongly expressed in the hemocytes and ovary of E. carinicauda. Ecß-actin mRNA expression varied during ovarian development, with high levels observed at stages I and V. Therefore, caution should be taken when using the Ecß-actin gene as an endogenous control gene.

Meta-analysis of IL-6 -174G/C polymorphism and psoriasis risk.

Previous studies examining the association between interleukin-6 (IL-6) -174G/C polymorphism and psoriasis risk have produced inconsistent results. The aim of this study was to offer a comprehensive review of the association between IL-6 -174G/C polymorphism and psoriasis risk through a meta-analysis. Literature search of PubMed and Embase databases was conducted to identify all eligible studies published before October 29, 2015. Four case-control studies involving 651 psoriasis cases and 552 controls were included in this meta-analysis. Data were extracted, and pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to assess the associations. Combined analysis revealed a significant association between this polymorphism and psoriasis risk under the recessive model (OR = 1.69, 95%CI = 1.12-2.55, P = 0.013 for GG vs GC + CC), and the heterozygous comparison model (OR = 1.70, 95%CI = 1.29-2.23, P < 0.001 for GG vs GC). However, no significant association was observed under the allelic model (OR = 1.37, 95%CI = 0.99-1.89, P = 0.060 for G vs C), the dominant model (OR = 1.25, 95%CI = 0.92-1.71, P = 0.152 for GG + GC vs CC), and the homozygote comparison model (OR = 1.62, 95%CI = 0.79-3.32, P = 0.186 for GG vs CC). We conclude that the IL-6 -174G/C polymorphism contributes to psoriasis risk. However, further studies should be performed to validate our results.

Risk factors of node metastasis in cervical carcinoma.

Purpose ofinvestigation: To investigate the metastatic risk factors of pelvic lymph nodes in patients with cervical carcinoma in Stage Ia2 and IIa2. MATERIALS AND METHODS: The clinic pathologic parameters in 337 patients with Stage Ia2-IIa2 cervical carcinoma were retrospectively analyzed. The risk factors for pelvic lymph node metastasis were evaluated by the way of univariate X2 statistic analysis and binary logistic regression analysis. RESULTS: The lymph nodes metastasis rate was 11.87% (40/337). Single variable analysis showed that age, clinical stage, the size of tumor ≥ four cm, depth of stromal invasion 2/3, lymph-vascular space involvement (LVSI), and parametrial extension were related to the metastasis of lymph nodes. Multivariate analysis showed that the size of tumor, depth of stromal invasion, LVSI, and parametrial extension were independent risk factors. CONCLUSION: Patients with tumor size ≥ four cm, stromal invasion ≥ 2/3, LVSI, and parametrial extension were at high risk of lymph node metastasis.

Identification and expression analysis of the sting gene, a sensor of viral DNA, in common carp Cyprinus carpio.

Stimulator of interferon gene (sting) was identified and characterized from common carp Cyprinus carpio. The sting messenger (m)RNA encoded a polypeptide of 402 amino acids with a calculated molecular mass of 46·184 kDa and an isoelectronic point of 6·08. The deduced protein of sting contained a signal peptide, three transmembrane motifs in the N-terminal region and four putative motifs (RXR) found in resident endoplasmic reticulum proteins. mRNA expression of sting was present in twelve investigated tissues, and was up-regulated by koi herpesvirus (KHV) in vivo and in vitro. The transcription of sting was altered by poly(I:C) and poly(dT:dA) stimulation in vitro. The findings suggested that sting is an inducible gene involved in innate immunity against DNA- and RNA-derived pathogens. To investigate defence mechanisms in C. carpio development, sting level in embryos, larvae and juvenile fish was monitored following KHV challenge. The sting message was negligible in embryos prior to hatching, but observed at higher transcriptional levels throughout larval and juvenile stages. Investigation showed the mRNA expression profiles of genes encoding for proteins promoting various functions in the interferon pathway, from pattern recognition receptors to antiviral genes, to be significantly induced in all examined organs by in vivo infection with KHV. Following KHV infection, the ifn message was significantly downregulated in spleen, head kidney, brain and hepatopancreas but notably up-regulated in gill, intestine and skin, suggesting that ifn induction might be related to the mucosal immune system and virus anti-ifn mechanisms. These results provided the basis for further research into the role and mechanisms of sting in fishes.

[The effect and factors associated with outcome of extracorporeal membrane oxygenation in refractory cardiogenic shock patients].

Objective: To observe the clinical efficacy and factors associated with outcome of extracorporeal membrane oxygenation (ECMO) in refractory cardiogenic shock patients. Methods: Patients with refractory cardiogenic shock received ECMO treatment in our hospital from May 2013 to November 2015 were retrospectively analyzed. The clinical status before ECMO support, ECMO timing, complications and outcome were observed and analyzed.The hemodynamic data and the amount of vasoactive drugs at 2 hours before ECMO support and at 2, 6, 24 and 48 hours after ECMO support were collected and compared. Results: Ten refractory cardiogenic shock patients were included in this study (5 acute fulminant myocarditis patients, 4 acute myocardial infarction patients, 1 myocardial rupture patient (6 males, 4 females, age ranged 12 to 56 years). Before ECMO, the mean left ventricular ejection fraction (LVEF) was (31.4±10.2)%, the mean score of APACHE Ⅱ was 26.6±10.8. Eight patients developed cardiac arrests and the duration of CPR ranged from 10 to 300 minutes and three patients received IABP. CVP decreased, BP increased, HR decreased, ScVO2 increased, dose of dobutamine decreased at 2 hours after ECMO support. After ECMO support for 6 hours, lactate decreased, dose of norepinephrine decreased. After ECMO support for 24 and 48 hours, hemodynamics became stable and shock was significantly improved. Complication including infection of limb and catheterization site occurred in 3 patients, femoral arterial thrombosis occurred in 2 patients, critical limb ischemia occurred in 2 patients, hemorrhage at the catheterization site occurred in 2 patients. The duration of ECMO ranged from 2 to 220 hours. Nine patients could be weaned off ECMO support and 6 patients survived to hospital discharge. Two patients died due to too late ECMO support, the other two patients died due to severe complication of limb. Conclusions: ECMO can rapidly improve hemodynamic stability of patients with cardiogenic shock. Accurate assessing the timing of ECMO support and decreasing complication of limb play a critical role on improving outcome in refractory cardiogenic shock patients.

Association between IL-21 polymorphism and systemic lupus erythematosus: a meta-analysis.

Several case-control studies have been conducted to investigate the association between Interleukin-21 (IL-21) polymorphisms and systemic lupus erythematosus (SLE) susceptibility, and most of the studies focused on IL-21 rs907715 and rs2221903 polymorphisms. Given the inconsistent results from these studies, the present meta-analysis aimed to obtain a more precise estimate of the association between IL-21 rs907715 and rs2221903 polymorphisms and SLE. Studies regarding these specific polymorphisms and SLE were retrieved from PubMed, Embase, Web of Science, CNKI, and CBM. Data were extracted and meta-analysis was performed using the STATA 12.0 software. For the IL-21 rs907715 polymorphism, seven sets of comparisons involving 7977 SLE cases and 8097 healthy controls were considered. Results showed that there were significant differences in the IL-21 rs907715 genotype distribution between SLE patients and healthy controls in the comparisons of all genetic models. Upon stratified analysis by ethnicity, a similar result was found in the Caucasian and African-American population. For the IL-21 rs2221903 polymorphism, seven sets of comparisons involving 7990 SLE cases and 8098 healthy controls were considered. Results showed that there were significant differences in the IL-21 rs2221903 genotype distribution between SLE patients and healthy controls in the comparisons of GG versus AA and GG versus GA+AA. Upon stratified analysis by ethnicity, a similar result was found in the Caucasian population. This meta-analysis suggests that the both IL-21 rs907715 and rs2221903 polymorphisms may be associated with SLE susceptibility. As current evidence remains limited, further studies are needed to warrant the association between IL-21 rs907715 and rs2221903 polymorphisms and SLE susceptibility.

Association between MTHFR 677C/T polymorphism and psoriasis risk: a meta-analysis.

Previous studies investigating the association between methylenetetrahydrofolate reductase (MTHFR) 677C/T polymorphisms and psoriasis risk have reported inconsistent results. The present meta-analysis aimed to comprehensively evaluate the association between MTHFR 677C/T polymorphism and psoriasis risk. The studies regarding the association between MTHFR 677C/T polymorphism and psoriasis risk were retrieved from the PubMed, Embase, Web of Science, Chinese National Knowledge Infrastructure, and Chinese Biomedical databases. Data were extracted and statistical analysis was performed with the program STATA 12.0. A total of seven studies involving 1290 psoriasis cases and 1068 healthy controls were retrieved. Combined analysis showed that there was no significant difference in MTHFR 677C/T genotype distribution between psoriasis and control subjects in the comparisons C vs T, CC vs CT + TT, CC + CT vs TT, CC vs TT, and CC vs CT [respectively: odds ratio (OR) = 0.98, 95% confidence interval (CI) = 0.76-1.26, P = 0.882; OR = 1.11, 95%CI = 0.81-1.51, P = 0.526; OR = 0.79, 95%CI = 0.53-1.19, P = 0.261; OR = 0.88, 95%CI = 0.51-1.52, P = 0.648; OR = 1.19, 95%CI = 0.90-1.58, P = 0.217]. Subgroup analysis by ethnicity also showed no significant association between MTHFR 677C/T polymorphism and psoriasis risk in both Asian and Caucasian populations. In conclusion, this meta-analysis indicates that MTHFR 677C/T polymorphism may not be associated with psoriasis risk.

Meta-analysis of the TNF-α-308G/A polymorphism and vitiligo risk.

Several case-control studies have been conducted to investigate the association between the tumor necrosis factor-α (TNF-α)-308G/A polymorphism and vitiligo risk. However, the results of these studies are inconsistent; therefore, we attempted to comprehensively evaluate the association between TNF-α-308G/A polymorphism and vitiligo risk via a meta-analysis. Studies reporting the association between TNF-α-308G/A polymorphism and vitiligo risk were retrieved from PubMed and EmBase databases. Data were extracted from these studies and the pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to assess the association. Six case-control studies including 1391 vitiligo cases and 2455 control subjects were included in this meta-analysis. The overall results showed the lack of a significant difference in TNF-α-308G/A genotype distribution between the patients and controls when the G allele and GG, GG + GA, GG, and GG genotypes were compared against the A allele and the GA + AA, AA, AA, and GA genotypes, respectively (ORs = 0.65, 0.53, 0.63, 0.41, 0.55; 95%CI = 0.35-1.23, 0.24-1.18, 0.10-4.09, 0.08-1.97, 0.25-1.21; P = 0.188, 0.121, 0.627, 0.264, 0.135, respectively). This meta-analysis suggests that the TNF-α-308G/A polymorphism may not be associated with vitiligo risk. As few studies are available in this field and current evidence remains limited, these results must be corroborated with well-designed and larger studies in the future.

Identification and expression of the laboratory of genetics and physiology 2 gene in common carp Cyprinus carpio.

In this study, a laboratory of genetics and physiology 2 gene (lgp2) from common carp Cyprinus carpio was isolated and characterized. The full-length complementary (c)DNA of lgp2 was 3061 bp and encoded a polypeptide of 680 amino acids, with an estimated molecular mass of 77 341·2 Da and a predicted isoelectric point of 6·53. The predicted protein included four main overlapping structural domains: a conserved restriction domain of bacterial type III restriction enzyme, a DEAD-DEAH box helicase domain, a helicase super family C-terminal domain and a regulatory domain. Real-time quantitative polymerase chain reaction (PCR) showed widespread expression of lgp2, mitochondrial antiviral signalling protein (mavs) and interferon transcription factor 3 (irf3) in tissues of nine organs. lgp2, mavs and irf3 expression levels were significantly induced in all examined organs by infection with koi herpesvirus (KHV). lgp2, mavs and irf3 messenger (m)RNA levels were significantly up-regulated in vivo after KHV infection, and lgp2 transcripts were also significantly enhanced in vitro after stimulation with synthetic, double-stranded RNA polyinosinic polycytidylic [poly(I:C)]. These findings suggest that lgp2 is an inducible protein involved in the innate immune defence against KHV in C. carpio. These results provide the basis for further research into the role and mechanisms of lgp2 in fishes.

HTRA3 is reduced in ovarian cancers regardless of stage.

Ovarian cancer is the leading cause of death in gynaecological cancers. The high temperature requirement factor A3 (HtrA3) is involved in the pathogenesis of ovarian cancer. In this study we investigated whetherHtrA3 protein levels were altered in subtypes of ovarian cancer and whether HtrA3 down-regulation was associated with peritoneal metastasis. Ovarian cancer tissues from 89 patients were analyzed by immunohistochemistry. The levels of HtrA3 protein were lower in all subtypes of ovarian cancer and the lowest levels of HtrA3 were in epithelial ovarian cancer. The down-regulation of HtrA3 levels was not correlated with peritoneal metastasis of epithelial ovarian cancer.

Accumulation and elimination of enrofloxacin and its metabolite ciprofloxacin in the ridgetail white prawn Exopalaemon carinicauda following medicated feed and bath administration.

Accumulation and elimination of enrofloxacin and its metabolite ciprofloxacin were evaluated in Exopalaemon carinicauda following medicated feed at dose of 10 mg/kg weight body per day for five consecutive days and 10 mg/L bath for five consecutive days at 18 °C. At different times, nine ridgetail white prawns were randomly selected from the tank and sampled after the last medicated feed or bath administration. The concentration of enrofloxacin and ciprofloxacin in the main tissues (hepatopancreas, muscle, gill, and ovary) was detected by HPLC. The results showed that the maximum concentrations of enrofloxacin were 3.408 ± 0.245, 0.554 ± 0.088, 0.789 ± 0.074, and 0.714 ± 0.123 µg/g for hepatopancreas, muscle, gill, and ovary, respectively, at 1 day after the last medicated feed treatment. The enrofloxacin concentrations were 2.389 ± 0.484, 0.656 ± 0.012, 0.951 ± 0.144, and 3.107 ± 0.721 µg/g in hepatopancreas, muscle, gill, and ovary, respectively, at 1 day after the last bath administration. Ciprofloxacin could be detected in hepatopancreas, muscle, gill, and ovary. However, the concentrations of ciprofloxacin were much lower in comparison with that of enrofloxacin in various tissues. The concentrations of enrofloxacin plus ciprofloxacin in hepatopancreas, muscle, gill, and ovary followed an eliminating pattern during the sampling time after the two routes of administration. Based on data derived from this study, to avoid the enrofloxacin and ciprofloxacin residue in E. carinicauda, it should take at least 20 and 25 days to wash out the drug from the tissues after the last medicated feed and bath administration with enrofloxacin, respectively. These results helped the Chinese fishery department to lay down the current guidelines on enrofloxacin plus ciprofloxacin withdrawal periods for farmed shrimp.

A placental model of SARS-CoV-2 infection reveals ACE2-dependent susceptibility and differentiation impairment in syncytiotrophoblasts.

SARS-CoV-2 infection causes COVID-19. Several clinical reports have linked COVID-19 during pregnancy to negative birth outcomes and placentitis. However, the pathophysiological mechanisms underpinning SARS-CoV-2 infection during placentation and early pregnancy are not clear. Here, to shed light on this, we used induced trophoblast stem cells to generate an in vitro early placenta infection model. We identified that syncytiotrophoblasts could be infected through angiotensin-converting enzyme 2 (ACE2). Using a co-culture model of vertical transmission, we confirmed the ability of the virus to infect syncytiotrophoblasts through a previous endometrial cell infection. We further demonstrated transcriptional changes in infected syncytiotrophoblasts that led to impairment of cellular processes, reduced secretion of HCG hormone and morphological changes vital for syncytiotrophoblast function. Furthermore, different antibody strategies and antiviral drugs restore these impairments. In summary, we have established a scalable and tractable platform to study early placental cell types and highlighted its use in studying strategies to protect the placenta.

Metabolic regulation of polyamines and γ-aminobutyric acid in relation to spermidine-induced heat tolerance in white clover.

Heat stress decreases crop growth and yield worldwide. Spermidine (Spd) is a small aliphatic amine and acts as a ubiquitous regulator for plant growth, development and stress tolerance. Objectives of this study were to determine effects of exogenous Spd on changes in endogenous polyamine (PA) and γ-aminobutyric acid (GABA) metabolism, oxidative damage, senescence and heat shock protein (HSP) expression in white clover subjected to heat stress. Physiological and molecular methods, including colorimetric assay, high performance liquid chromatography and qRT-PCR, were applied. Results showed that exogenous Spd significantly alleviated heat-induced stress damage. Application of Spd not only increased endogenous putrescine, Spd, spermine and total PA accumulation, but also accelerated PA oxidation and improved glutamic acid decarboxylase activity, leading to GABA accumulation in leaves under heat stress. The Spd-pretreated white clover maintained a significantly higher chlorophyll (Chl) content than untreated plants under heat stress, which could be related to the roles of Spd in up-regulating genes encoding Chl synthesis (PBGD and Mg-CHT) and maintaining reduced Chl degradation (PaO and CHLASE) during heat stress. In addition, Spd up-regulated HSP70, HSP70B and HSP70-5 expression, which might function in stabilizing denatured proteins and helping proteins to folding correctly in white clover under high temperature stress. In summary, exogenous Spd treatment improves the heat tolerance of white clover by altering endogenous PA and GABA content and metabolism, enhancing the antioxidant system and HSP expression and slowing leaf senescence related to an increase in Chl biosynthesis and a decrease in Chl degradation during heat stress.

[Analysis of drug - resistant gene polymorphisms in Plasmodium falciparum imported from Equatorial Guinea to Shandong Province in 2015 and 2016].

OBJECTIVE: To investigate the drug-resistant gene polymorphisms in Plasmodium falciparum imported from Equatorial Guinea to Shandong Province. METHODS: From 2015 to 2016, blood samples were collected from imported P. falciparum malaria patients returning from Equatorial Guinea to Shandong Province, and genome DNA of the malaria parasite was extracted. The drug-resistant Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, and K13 genes of P. falciparum were amplified using a PCR assay, followed by DNA sequencing, and the sequences were aligned. RESULTS: The target fragments of all 5 drug-resistant genes of P. falciparum were successfully amplified and sequenced. There were 72.8%, 18.6%, and 8.6% of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfcrt gene, respectively, and all mutant haplotypes were CVIET (the underline indicates the mutation site). There were 20.0%, 61.4% and 18.6% of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfmdr1 gene, respectively, and the mutant haplotypes mainly included YF and NF (the underlines indicate the mutation sites). There were 1.4%, 98.6%, and 0 of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfdhfr gene, respectively, and AIRNI was the predominant mutant haplotype (the underline indicates the mutation site). There were 1.4%, 94.3%, and 4.3% of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfdhps gene, respectively, and SGKAA was the predominant mutant haplotype (the underline indicates the mutation site). The complete drug-resistant IRNGE genotype consisted of 8.6% of the Pfdhfr and Pfdhps genes, and the K13 gene A578S mutation occurred in 1.4% of the parasite samples. CONCLUSIONS: There are mutations in the Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, and K13 genes of P. falciparum imported from Equatorial Guinea to Shandong Province, with a low frequency in the Pfcrt gene mutation and a high frequency in the Pfmdr1, Pfdhfr, and Pfdhps gene mutations, and the K13 gene A578S mutation is detected in the parasite samples.