[Drug-resistant gene polymorphisms in Plasmodium falciparum isolated from Bioko Island, Equatorial Guinea in 2018 and 2019].

OBJECTIVE: To investigate the genetic polymorphisms of Plasmodium falciparum multidrug resistance protein 1 (PfMDR1), chloroquine resistance transporter (PfCRT) and Kelch 13 (PfK13) genes in Bioko Island, Equatorial Guinea, so as to provide insights into the development of the malaria control strategy in local areas. METHODS: A total of 85 peripheral blood samples were collected from patients with Plasmodium falciparum infections in Bioko Island, Equatorial Guinea in 2018 and 2019, and genomic DNA was extracted. The PfMDR1, PfCRT and PfK13 genes were amplified using a nested PCR assay. The amplification products were sequenced, and the gene sequences were aligned. RESULTS: There were no mutations associated with artemisinin resistance in PfK13 gene in Bioko Island, Equatorial Guinea, while drug-resistant mutations were detected in PfMDR1 and PfCRT genes, and the proportions of PfMDR1_N86Y, PfMDR1_Y184F and PfCRT_K76T mutations were 35.29% (30/85), 72.94% (62/85) and 24.71% (21/85), respectively. CONCLUSIONS: There are mutations in PfMDR1, PfCRT and PfK13 genes in P. falciparum isolates from Bioko Island, Equatorial Guinea.

Sleep and hypertension.

PURPOSE: Hypertension is a global public issue, and sleep status was regarded as its risk factor; however, the results were inconsistent. This study aims to deeply investigate the correlation between sleep status and hypertension. METHODS: The electronic databases Cochrane Library, Pubmed, and Embase updated to May 31, 2019, were retrieved. Studies were selected according to the predefined screening criteria, and their qualities were assessed by using quality check scales. Based on Stata 15.1 software, the associations between sleep status and hypertension were analyzed by meta-analyses, using odds ratio and 95% confidence interval as effect indexes. Furthermore, publication bias and small study bias were evaluated using Begg and Egger's test. In addition, sensitivity analysis was conducted through ignoring one study per time and then observing its influences on the pooled results. RESULTS: A total of 54 studies (involving 1,074,207 subjects) were eligible for this meta-analysis. Six factors were included in this study. Raised blood pressure was associated with obstructive sleep apnea (OSA), oxygen desaturation index (ODI), short sleep duration, and long sleep duration. The differences in ≤ 5 h, 6 h, ≥ 9 h, and 10 h groups had statistical significances, while there was no significant difference in ≥ 8 h group. Snoring is a risk factor of hypertension (OR = 1.94, 95%CI 1.41-2.67). Subgroup analysis was conducted and results were varied. CONCLUSIONS: The hypertension risk might be reduced by treated OSA, ODI, and snoring, as well as appropriate sleep duration. More studies with large sample sizes and high qualities should be included to support the findings further.

Application of miniSTR Loci and Its Detection System for Degraded Materials in Forensic Medicine.

OBJECTIVES: To establish multiplex system of 16 miniSTR loci, and explore its application value for the degraded materials in forensic medicine. METHODS: The multiplex system of 16 miniSTR loci was established using a six-dye fluorescence labeling technology and its application value in forensic medicine was assessed. RESULTS: A six-dye fluorescence labeling miniSTR amplification kit was developed, which enabled 15 autosomal STR loci, Amelogenin locus and DYS391 to be typed simultaneously. This method showed good specificity and could provide stable and accurate typing results with a sensitivity of 50 pg. This system also provided a good test result for the normal biological sample of actual cases. CONCLUSIONS: The multiplex system of 16 miniSTR loci has application value for degraded and trace materials with the advantages of high sensitivity and database compatibility, which can be used for forensic casework.

MOTS-c improves osteoporosis by promoting osteogenic differentiation of bone marrow mesenchymal stem cells via TGF-ß/Smad pathway.

OBJECTIVE: To explore whether MOTS-c could improve osteoporosis by promoting osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs) via transforming growth factor-ß (TGF-ß)/Smad pathway. MATERIALS AND METHODS: Rat BMSCs were isolated and cultured, followed by osteogenic and lipid differentiation. CCK-8 (cell counting kit-8) assay was performed to detect the highest treatment dose of MOTS-c that did not affect cell proliferation. Expressions of osteogenesis-related genes (ALP, Bglap, and Runx2) were detected by qRT-PCR (quantitative Real-Time Polymerase Chain Reaction) and Western blot, respectively. Alizarin red staining and alkaline phosphatase (ALP) cytochemical staining were carried out to evaluate the effect of MOTS-c on BMSCs osteogenesis. TGF-ß/Smad pathway-related genes (TGF-ß1, TGF-ß2, and Smad7) in BMSCs treated with MOTS-c were detected. Finally, TGF-ß1 was knocked down to investigate the regulatory effect of MOTS-c on BMSCs osteogenesis. RESULTS: BMSCs exhibited an elongated morphology and was identified with a high purity by flow cytometry. After osteogenic differentiation, alizarin red staining and ALP staining were all positive. MOTS-c treatment could remarkably stimulate the formation of calcified nodules in BMSCs. Besides, TGF-ß/Smad pathway-related genes were significantly upregulated after BMSCs were treated with MOTS-c. Promoted osteogenesis by MOTS-c treatment was reversed by the TGF-ß1 knockdown. CONCLUSIONS: MOTS-c promotes cell differentiation of BMSCs to osteoblasts via TGF-ß/Smad pathway.

[Recurrent bleeding following the renal artery embolization treating post-percutaneous nephrolithotomy hemorrhage: causes and countermeasure].

Objective: To explore the causes and countermeasure in recurrent bleeding following the selective renal artery embolization treating post-percutaneous nephrolithotomy hemorrhage. Methods: A total of 334 patients of severe renal hemorrhage associated with percutaneous nephrolithotomy (PCNL) from March 2011 to April 2015 were analyzed retrospectively.All the patients underwent super selective angiography and renal artery embolization.The causes of recurrent hemorrhage were analyzed and principles for diagnosis and embolization were studied. Results: The initial embolization was performed in 329 cases hospitalized in the First Affiliated Hospital of Guangzhou Medical University and 318 cases were successfully stopped bleeding with a hemostatic rate of 96.7 %(318/329). Of total 334 consecutive cases, there were 16 cases of recurrent renal hemorrhage, 11 cases were initially embolized in this hospital, and otherwise the other 5 cases were in other hospitals. Causes of recurrent hemorrhage were missed embolization of tiny pseudoaneurysm (n=12), and two cases of 12, the tiny pseudoaneurysm were feeding by accessory renal arteries, undetected arteriovenous fistula(n=2), recanalization of the embolized arteries (n=2). Conclusion: The causes of recurrent bleeding fallowing the initial selective renal artery embolization treating post-percutaneous nephrolithotomy hemorrhage are varied, and missed embolization of tiny pseudoaneurysm is the major cause of unsuccessful initial renal artery embolization. To strengthen the understanding of tiny pseudoaneurysm is helpful to improve the success rate of hemostasis.

[Langerhans cell histiocytosis of the thyroid: a case report].

Langerhans cell histiocytosis (LCH) is a rare monoclonal disease,its clinical presentation is highly variable because it can affect multiple organs, such as lung, bone, skin, lymph nodes, hypothalamopituitary axis, and other multiple sites. LCH involving the thyroid gland is extremely rare, here we reported a rare case of LCH involving thyroid, presenting as painless thyroid goiters.

[New progress of BRAF gene and thyroid cancer].

It is noteworthy that the incidence of thyroid cancer around the world has increased significantly in recent decades, raising an imperative need to research its pathogenesis, diagnosis and treatment. Up to now, fine needle aspiration biopsy (FNAB) of thyroid has been acknowledged to discriminate benign from malignant thyroid nodules with the highest sensitivity and specificity. However, 10% to 40% thyroid nodules cannot be discriminated by FNAB. Therefore, it is vitally important to look for highly-correlated tumor makers in molecule level. BRAF mutation is a focus in thyroid cancer research, and some studies showed that this mutation is essential to the onset and development of thyroid cancer, especially papillary thyroid cancer. Joint detection of BRAF mutation could improve diagnostic sensitivity of thyroid cancer, which is crucial for thyroid cancer diagnosis and classification. As for treatment, the discovery of target gene enabled molecule therapy for thyroid cancer, raising hopes for patients with thyroid cancer that refractory to conventional treatments. Currently, many molecule therapeutics relating to BRAF has already undergone clinical trials. It is believed that further research on BRAF-thyroid cancer relationship could create a new field for diagnosis and treatment of thyroid cancer, and set a mode for discovering others molecule markers.

Baicalin can attenuate the inhibitory effects of mifepristone on Wnt pathway during peri-implantation period in mice.

Wnts are considered as important factors in uterus developmental process and embryo implantation. Baicalin has been demonstrated to possess tocolytic properties. In order to investigate the effect of baicalin on the Wnt signaling pathway during the peri-implantation, pregnant Kuming mice were randomly divided into four groups: control group, baicalin group administered with 40mg/kg BW of baicalin through an intragastric gavage on day 2 to 7 of the pregnancy (Pd2-Pd7), mifepristone group treated with 4mg/kg BW of mifepristone, an abortifacient agent, via subcutaneous administration on Pd4, and baicalin+mifepristone group treated with their combination. The concentrations of the implantation-related steroid hormones (progesterone and estradiol) in the blood serum were measured with RIA. The gene and protein expression levels of the important molecules of the Wnt pathway (Wnt4, LRP6, Dkk1 and ß-catenin) in the endometrium were detected with RT-PCR and western blot, respectively. The results showed that baicalin decreased (P<0.05) the estradiol levels on Pd4-Pd8 and increased (P<0.05) the progesterone levels on Pd3-Pd8. Mifepristone increased (P<0.05) the estradiol levels on Pd5-Pd8 and decreased (P<0.05) the progesterone levels on Pd6-Pd8. Compared with the control group, baicalin increased the gene and protein expression levels of Wnt4, LRP6 and ß-catenin (P<0.05) and decreased the gene and protein expression levels of Dkk1 (P<0.05) during the middle-to-late stage of the experiment in mice uterine tissue. Baicalin alleviated the mifepristone-induced increase or decrease in the serum levels of progesterone and estradiol, and the gene or protein expression levels of Wnt4, LRP6 and ß-catenin. The tocolytic properties tocolysis of baicalin may be realized through regulating the levels of estrogen/progesterone and the important components of canonical Wnt signaling pathway during the embryo implantation process intervened with the subcutaneous administration of mifepristone in the mice.

Processing of facial expressions of emotions in healthy volunteers: an exploration with event-related potentials and personality traits.

AIMS OF THE STUDY: Previous studies have shown that event-related potentials (ERPs) are modulated by anxiety or psychopathic personality traits. Therefore, we hypothesized that the automatic processing of facial expressions of emotions (FEE) is also correlated with related disordered personality traits. METHODS: Thirty-seven healthy volunteers underwent both an "oddball" ERP recording to facial expressions of Anger, Happiness, Sadness, and Neutral, and a test of the Dimensional Assessment of Personality Pathology (DAPP). RESULTS: Mean reaction time was longer in response to anger than to other facial expressions. Facial expressions of Anger, Happiness and Sadness did not affect N1 (N170). By contrast, Happiness elicited a delayed P2, Anger elicited both a smaller N2 and a delayed P3b, and both Happiness and Anger elicited a P3b of higher amplitude. In addition, P3a latencies to Happiness were negatively correlated with DAPP Identity problems, and P3b latencies to Happiness were negatively correlated with DAPP Stimulus seeking, Callousness, Passive aggressivity, and Narcissism. CONCLUSION: Our study demonstrates that Anger implicitly captures attentional resources, and Happiness triggers more facilitated processing in individuals with dissocial traits.

Targeted inhibition of platelet-derived growth factor receptor-beta subunit in hepatic stellate cells ameliorates hepatic fibrosis in rats.

The activation of hepatic stellate cells (HSCs) is the key event of the pathogenesis of hepatic fibrosis. Platelet-derived growth factor (PDGF) is the most potent mitogen for HSCs, and PDGF receptor-beta subunit (PDGFR-beta) is required for the proliferation of HSCs induced by PDGF. In this study, a high gene-silencing-efficacy PDGFR-beta small interference RNA (siRNA) was synthesized that could suppress the PDGFR-beta expression and inhibit the activation and proliferation but could not induce the apoptosis of HSCs in vitro. To avoid the side effect of nonspecific interference of PDGFR-beta, we constructed an HSCs-specific short hairpin RNA (shRNA) expression plasmid in which PDGFR-beta shRNA was driven by a glial fibrillary acidic protein (GFAP) promoter. The double-staining immunofluorescence examination indicated that GFAP promoter could target the transgene expression into HSCs in carbon tetrachloride induced acute injured rat's liver and bile duct ligation (BDL)-induced chronic injured rat's liver. Furthermore, HSCs-specific PDGFR-beta shRNA could relieve liver injury and hepatic fibrosis in the rat's model induced by BDL. This study demonstrates that PDGFR-beta siRNA may be presented as an antifibrogenic agent. The application of HSCs-specific RNA interference induced by the GFAP promoter might supply a new powerful tool for cell-specific gene therapy of hepatic fibrogenesis.

Toxic Effects in Siganus oramin by dietary exposure to 4-tert-octylphenol.

The integrated toxicities of 4-tert-octylphenol (t-OP) on Siganus oramin were investigated by dietary administration at doses of 5, 25 and 125 mg/kg body weight over 28 days. Significant increase was observed in the activity of hepatic glutathione S-transferase at 125 mg/kg on both day 14 and 28 in males, and at all doses on day 28 in females, and in hepatosomatic index at 25 mg/kg on day 14 in both sexes. Plasma levels of testosterone and cortisol decreased significantly at all doses on day 28. Histopathologic changes in liver, spleen, intestine and testis deteriorated with increasing doses and duration. The results suggest that S. oramin is sensitive to t-OP, and the above endpoints may be potential biomarkers for evaluating toxicities of environmental pollutants such as t-OP.

Binding modes of two highly potent and nontoxic inhibitors of HIV-1 integrase.

The complex structures of Human Immunodeficiency Virus Type 1 (HIV-1) integrase binding two highly potent and nontoxic inhibitors, lithospermic acid (M/sub 5/22) and lithospermic acid B (M/sub 5/32), were obtained using docking calculations. Docking results provided detailed information of their binding modes. The binding sites of M/sub 5/22 and M/sub 5/32 were similar to the inhibitor 5-CITEP. The lowest docking energies for HIV-1 integrase binding M/sub 5/22 and M/sub 5/32 are in agreement with their corresponding lower IC/sub 50/ values. Our results on the chemical structure difference between M/sub 5/22 and M/sub 5/32 show that the carboxyl and hydroxyl groups on the side-chain of M/sub 5/32 are important chemical groups which could help to increase the effect against HIV-1 IN replication.

A randomised clinical trial of high-intensity focused ultrasound ablation for the treatment of patients with localised breast cancer.

High-intensity focused ultrasound (HIFU) is a noninvasive treatment that induces complete coagulative necrosis of a tumour at depth through the intact skin. This study was to explore the possibility of using HIFU for the treatment of patients with localised breast cancer in a controlled clinical trial. A total of 48 women with biopsy-proven breast cancer (T(1-2), N(0-2), M0) were randomised to the control group in which modified radical mastectomy was performed, and the HIFU group in which an extracorporeal HIFU ablation of breast cancer was followed by modified radical mastectomy. Short-term follow-up, pathologic and immunohistochemical stains were performed to assess the therapeutic effects on tumour and complications of HIFU. The results showed that no severe side effect was observed in the HIFU-treated patients. Pathologic findings revealed that HIFU-treated tumour cells underwent complete coagulative necrosis, and tumour vascular vessels were severely damaged. Immunohistochemical staining showed that no expression of PCNA, MMP-9, and CD44v6 was detected within the treated tumour cells in the HIFU group, indicating that the treated tumour cells lost the abilities of proliferation, invasion, and metastasis. It is concluded that, as a noninvasive therapy, HIFU could be effective, safe, and feasible in the extracorporeal treatment of localised breast cancer.

Killing effects of ganciclovir on human pulmonary adenocarcinoma cell A549 transduced with HSV1-TK gene in vitro and in vivo.

AIM: To observe the killing effects of ganciclovir (GCV) on the human pulmonary adenocarcinoma cell A549 transduced with Herpes simplex virus I type thymine kinase (HSV1-TK) gene in vitro and in vivo. METHODS: A retroviral vector containing the TK gene was constructed and transduced into a pulmonary carcinoma cell A549 by electroporation, to observe the sensitivity of the transfected cell to GCV in vitro and the bystander effect (MTT assay). Tumor cell apoptosis caused by the TK/GCV system was observed with a flow cell meter (FCM) and a scan electronic microscope (SEM). Recombination and expression of the TK gene were examined with DNA PCR and in situ hybridization, respectively. The therapeutic effect of GCV on subcutaneous tumor growth between transfected and parental cells was also compared. RESULTS: The sensitivity of the transfected cell to GCV was 46 times higher than that of the parental cell, and the bystander effect was stronger in high cell density than in low cell density. The subG0G1 peak was shown on the DNA histogram after A549-Tk cell was treated with 50 micromol/L GCV for 3 days by FCM, but not in the A549 cell. A cell cycle analysis showed that the apoptotic cell in the A549-TK and A549 cells were (12.2+/-1.7) % and (1.3 +/- 0.3) %, respectively (P < 0.01). The cell apoptosis features of nuclear condensation, apoptotic vesicle, and nuclear showing semimoon feature were found in the A549-TK cell by SEM, but not in the A549 cell. Recombination and expression of the TK gene were positive in the transfected cell. In vivo, the growth of tumors formed by the transfected cell was apparently inhibited by GCV, but not in the control group. CONCLUSION: The transfected cell obtained sensitivity to GCV and the bystander effect was closely related to intercellular touch. The TK/GCV system killing tumor cell was related to cell apoptosis. GCV inhibited the growth of tumors which were inoculated by A549-TK cell in vivo.

Pathological changes in human malignant carcinoma treated with high-intensity focused ultrasound.

The purpose of this study was to investigate the pathologic changes of extracorporeal ablation of human malignant tumors with high-intensity focused ultrasound (HIFU). HIFU treatment was performed in the 164 patients with liver cancer, breast cancer, malignant bone tumor, soft tissue sarcoma and other malignant tumors at focal peak intensities from 5000 W x cm(-2) to 20,000 W x cm(-2), with operating frequencies of 0.8 to 3.2 MHz. To explore the pathologic impact of extracorporeal HIFU, 30 patients with malignant carcinoma underwent surgical removal after HIFU treatment. Pathologic findings showed that the treated tissues demonstrated homogeneous coagulative necrosis with an irreversible tumor cell death and severe damage to tumor blood vessels at the level of microsvasculature within the HIFU-targeted region. Thermolesions to intervening tissue were never observed. The treated region had a sharp border comprising only several cell layers between the treated and untreated areas. The repair of lesions had the processes of necrotic tissue absorption and granulation tissue replacement. It is concluded that extracorporeal treatment of human solid malignancies with HIFU could be safe, effective and feasible. As a noninvasive therapy, HIFU would be used clinically to treat patients with solid malignancies.

Phenanthrenequinone-doped poly(methyl methacrylate) photopolymer bulk for volume holographic data storage.

We present the design and fabrication of a phenanthrenequinone-doped poly(methyl methacrylate) photopolymer material. Large blocks of samples were made, and the material showed negligible shrinkage after optical exposures. We recorded and reconstructed 250 holograms at a single spot, using a 1-cm(3) block.

Molecular dynamics simulations of the gramicidin A-dimyristoylphosphatidylcholine system with an ion in the channel pore region.

To investigate the process of ion permeation in an ion channel systematically, we performed molecular dynamics (MD) simulations on a gramicidin A (GA)-phospholipid model system with an ion in the channel pore region. Each of the three types of ions (Ca2+, Na+ Cl-) was placed at five different positions along the channel axis by replacing a water molecule. MD simulations were performed on each system at constant pressure and constant temperature. The MD trajectories showed that the Ca2+ and Na+ ions could stably fluctuate in the pore region, but the Cl- ion was pushed out because of the unfavorable interaction with the channel. This result is consistent with experimental data. It was also found that the conformation of the GA channel underwent a significant change due to the presence of the ion, and the two ends of the GA monomer were more flexible than its middle region. In particular, the dramatic change of local pore radius near the ion indicated this kind of deformation. The strong interaction between the ion and carbonyl oxygen atoms of GA was the major contributor to this change. Furthermore, it was found that the ethanolamine group of the GA molecule was the most flexible group in the GA channel and often observed to block the entrance of GA. These results imply that the deformation of channel structure plays a very important factor in ion permeation, and the ethanolamine group may play a key role in regulating ion entry into the pore. In conclusion, our results indicate that the ion has a dominant influence on the structure of the GA channel and that the flexibility of the ion channel is a crucial factor in the ion permeation process.

Inhibitory effect of Ginkgo biloba extract on the expression of inducible nitric oxide synthase in endothelial cells.

Excessive production of nitric oxide (NO) may have cytotoxic effects through the formation of peroxynitrite with superoxide. The extract of Ginkgo biloba leaves (EGb) has been demonstrated to be a potent scavenger of free radicals. Although EGb has been shown recently to inhibit NO production in macrophages, its effect on NO production in endothelial cells is largely unknown. The objective of this study was to elucidate the mechanism by which EGb affects NO production in a human endothelial cell line (ECV304). After cells were incubated with EGb (10-100 microg/mL) for 2 or 4 hr, the amounts of NO metabolites released by the cells were quantitated, and cellular NOS activities were determined following the conversion of [3H]arginine to [3H]citrulline. NOS protein expression was determined by western immunoblotting analysis. mRNA levels were examined by reverse transcription-polymerase chain reaction (RT-PCR) analysis. EGb (50 microg/mL) caused a 30% reduction of NO metabolites released by endothelial cells. Following EGb treatment, cellular inducible NO synthase (iNOS) activity was reduced by 28% with a concomitant reduction in the levels of iNOS protein mass and mRNA. There was no change in the activity or protein mass of constitutive NO synthase in these cells. EGb inhibited NO production by attenuating the level of iNOS mRNA in ECV304 cells. Selective inhibition of iNOS by EGb may be therapeutically relevant in modulating NO production in endothelial cells.

Constructing the suitable initial configuration of the membrane-protein system in molecular dynamics simulations.

A method for constructing the suitable initial configuration of the membrane-protein system for molecular dynamics (MD) simulations is presented. This method could provide some hydrated initial configurations and help us to determine the best surface area of the system by contracting the surface area and comparing the optimized lowest energy of the system by energy minimization. The gramicidin A (GA) channel in;the fully hydrated dimyristoylphosphatidylcholine (DMPC) bilayer was used as our model. Three configurations with different surface areas were selected and applied for one 400 ps and two 300 ps MD simulations at constant pressure and temperature. All simulations were fairly stable without any constraints. Through analysis of the MD trajectories we found that the system with the best surface area was more stable than the other two systems, whose sizes were changed in the simulations. Further analysis of the bilayer normal length and the order parameters of the lipid alkyl tails indicates that the system with the best surface area shows some characteristics of the L(alpha) phase, while both the smaller and the larger size systems have distinct deviations from the L(alpha) phase that we expect. This illustrates that the correct surface area and the suitable initial configuration have an important influence on the phase of the membrane in the MD simulation. In addition, by comparing the root mean square differences of GA relative to the initial structure and interaction energy between different components of the system for all three systems, we find that the state of the DMPC bilayer has exerted a significant influence on the structure of GA. All these results demonstrate the validity of our method for constructing the initial configuration of the membrane-protein system for MD simulations.

[Variation of chemical constituents in processed licorice roots: quantitative determination of saponin and flavonoid constituents in bark removed and roasted licorice roots].

As a continuing study of chemical characterization of crude drug processing, we have been analyzing the chemical constituents in licorice roots before and after processing. At first, we analyzed chemical constituents in licorice roots of various origins. Next, we have developed the HPLC analytical method, by which saponins and flavonoids, major constituents in various licorice roots, were determined simultaneously in a quantitative manner. In this paper, by means of the HPLC analytical method, chemical constituents in licorice roots, processed and unprocessed, were determined. It was found that nonglycosidic flavonoid constituents were mostly lost while root bark removing, whereas, in roasted licorice roots, sugar chains in the saponin and glycosidic flavonoid constituents were hydrolyzed stepwise during roasting through hydrothermolysis.