Cinnamaldehyde: An effective component of Cinnamomum cassia inhibiting Helicobacter pylori.

ETHNOPHARMACOLOGICAL RELEVANCE: Cinnamomum cassia Presl (Cinnamomum cassia) is a common traditional Chinese medicine, which can promote the secretion and digestion of gastric juice, improve the function of gastrointestinal tract. Cinnamaldehyde (CA) is a synthetic food flavoring in the Chinese Pharmacopoeia. AIM OF THE STUDY: This study aimed to search for the active ingredient (CA) of inhibiting H. pylori from Cinnamomum cassia, and elucidate mechanism of action, so as to provide the experimental basis for the treatment of H. pylori infection with Cinnamomum cassia. MATERIALS AND METHODS: It's in vitro and in vivo pharmacological properties were evaluated based on minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and an acute gastric inflammation model in mice infected with H. pylori. Drug safety was evaluated using the CCK8 method and high-dose administration in mice. The advantageous characteristics of CA in inhibiting H. pylori were confirmed using acidic conditions and in combination with the antibiotics. The mechanism underlying the action of CA on H. pylori was explored using scanning electron microscopy (SEM), adhesion experiments, biofilm inhibition tests, ATP and ROS release experiments, and drug affinity responsive target stability (DARTS) screening of target proteins. The protein function and target genes were verified by molecular docking and Real-Time quantitative reverse transcription PCR (qRT-PCR). RESULTS: The results demonstrated that CA was found to be the main active ingredient against H. pylori in Cinnamomum cassia in-vitro tests, with a MIC of 8-16 µg/mL. Moreover, CA effectively inhibited both sensitive and resistant H. pylori strains. The dual therapy of PPI + CA exhibited remarkable in vivo efficacy in the acute gastritis mouse model, superior to the standard triple therapy. DARTS, molecular docking, and qRT-PCR results suggested that the target sites of action were closely associated with GyrA, GyrB, AtpA, and TopA, which made DNA replication and transcription impossible, then leading to inhibition of bacterial adhesion and colonization, suppression of biofilm formation, and inhibition ATP and enhancing ROS. CONCLUSIONS: This study demonstrated the suitability of CA as a promising lead drug against H. pylori, The main mechanisms can target GyrA ect, leading to reduce ATP and produce ROS, which induces the apoptosis of bacterial.

Nanoengineered Red Blood Cells Loaded with TMPRSS2 and Cathepsin L Inhibitors Block SARS-CoV-2 Pseudovirus Entry into Lung ACE2+ Cells.

The enzymatic activities of Furin, Transmembrane serine proteinase 2 (TMPRSS2), Cathepsin L (CTSL), and Angiotensin-converting enzyme 2 (ACE2) receptor binding are necessary for the entry of coronaviruses into host cells. Precise inhibition of these key proteases in ACE2+ lung cells during a viral infection cycle shall prevent viral Spike (S) protein activation and its fusion with a host cell membrane, consequently averting virus entry to the cells. In this study, dual-drug-combined (TMPRSS2 inhibitor Camostat and CTSL inhibitor E-64d) nanocarriers (NCs) are constructed conjugated with an anti-human ACE2 (hACE2) antibody and employ Red Blood Cell (RBC)-hitchhiking, termed "Nanoengineered RBCs," for targeting lung cells. The significant therapeutic efficacy of the dual-drug-loaded nanoengineered RBCs in pseudovirus-infected K18-hACE2 transgenic mice is reported. Notably, the modular nanoengineered RBCs (anti-receptor antibody+NCs+RBCs) precisely target key proteases of host cells in the lungs to block the entry of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), regardless of virus variations. These findings are anticipated to benefit the development of a series of novel and safe host-cell-protecting antiviral therapies.

Association between iron deficiency and prevalence of thyroid autoimmunity in pregnant and non-pregnant women of childbearing age: a cross-sectional study.

BACKGROUND: Thyroid autoimmunity (TAI) is prevalent among women of reproductive age and associated with adverse pregnancy outcomes. This study aimed to investigate the association between iron nutritional status and the prevalence of TAI in women during the first trimester of pregnancy and in non-pregnant women of childbearing age. METHODS: Cross-sectional analysis of 7463 pregnant women during the first trimester of pregnancy and 2185 non-pregnant women of childbearing age nested within the sub-clinical hypothyroid in early pregnancy study, a prospective collection of pregnant and non-pregnant women's data, was conducted in Liaoning province of China between 2012 and 2015. Serum thyrotropin, free thyroxine, thyroid peroxidase antibodies (TPOAbs), thyroglobulin antibodies (TgAbs), serum ferritin, and urinary iodine were measured. Iron deficiency (ID) was defined as serum ferritin <15 µg/L and iron overload (IO) was defined as ferritin >150 µg/L. TPOAb-positive was defined as >34 U/mL and TgAb-positive was defined as >115 U/mL. Multilevel logistic regression was conducted to examine the association between TAI and different iron nutritional status after adjusting for potential confounders. RESULTS: The prevalence of isolated TPOAb-positive was markedly higher in women with ID than those without ID, in both pregnant and non-pregnant women (6.28% vs. 3.23%, χ = 10.264, P = 0.002; 6.25% vs. 3.70%, χ = 3,791, P = 0.044; respectively). After adjusting for confounders and the cluster effect of hospitals, ID remained associated with TPOAb-positive in pregnant and non-pregnant women (odds ratio [OR]: 2.111, 95% confidence interval [CI]: 1.241-3.591, P = 0.006; and OR: 1.822, 95% CI: 1.011-3.282, P = 0.046, respectively). CONCLUSION: ID was associated with a higher prevalence of isolated TPOAbs-positive, but not with isolated TgAb-positive, in both pregnant women during the first trimester of pregnancy and non-pregnant women of childbearing age, while IO was not associated with either isolated TPOAb-positive or isolated TgAb-positive. CLINICAL TRIAL REGISTRATION: ChiCTR-TRC-13003805, http://www.chictr.org.cn/index.aspx.

Expression of pyruvate dehydrogenase is an independent prognostic marker in gastric cancer.

AIM: To investigate the expression and prognostic role of pyruvate dehydrogenase (PDH) in gastric cancer (GC). METHODS: This study included 265 patients (194 male, 71 female, mean age 59 years (range, 29-81 years) with GC who underwent curative surgery at the First Affiliated Hospital of China Medical University from January 2006 to May 2007. All patients were followed up for more than 5 years. Patient-derived paraffin embedded GC specimens were collected for tissue microarrays (TMAs). We examined PDH expression by immunohistochemistry in TMAs containing tumor tissue and matched non-neoplastic mucosa. Immunoreactivity was evaluated independently by two researchers. Overall survival (OS) rates were determined using the Kaplan-Meier estimator. Correlations with other clinicopathologic factors were evaluated by two-tailed χ(2) tests or a two-tailed t-test. The Cox proportional-hazard model was used in univariate analysis and multivariate analysis to identify factors significantly correlated with prognosis. RESULTS: Immunohistochemistry showed that 35.47% of total cancer tissue specimens had cytoplasmic PDH staining. PDH expression was much higher in normal mucosa specimens (75.09%; P = 0.001). PDH expression was correlated with Lauren grade (70.77% in intestinal type vs 40.0% in diffuse type; P = 0.001), lymph node metastasis (65.43% with no metastasis vs 51.09% with metastasis; P = 0.033), lymphatic invasion (61.62% with no invasion vs 38.81% with invasion; P = 0.002), histologic subtypes (70.77% in intestinal type vs 40.0% in diffuse type; P = 0.001) and tumor-node-metastasis (TNM) stage (39% in poorly differentiated vs 65.91% in well differentiated and 67.11% in moderately differentiated; P = 0.001) in GC. PDH expression in cancer tissue was significantly associated with higher OS (P < 0.001). The multivariate analysis adjusted for age, Lauren classification, TNM stage, lymph node metastasis, histological type, tumor size, depth of invasion and lymphatic invasion showed that the PDH expression in GC was an independent prognostic factor for higher OS (HR = 0.608, 95%CI: 0.504-0.734, P < 0.001). CONCLUSION: Our study indicated that PDH expression is an independent prognostic factor in GC patients and that positive expression of PDH may be predictive of favorable outcomes.

[Construction and identification of human PRMT1 gene eukaryotic vector].

AIM: To construct the eukaryotic recombinant expression plasmid of pcDNA3.1(+)-PRMT1. METHODS: Human PRMT1 cDNA was amplified by reverse transcription polymerase chain reaction (RT-PCR). After digested by BamH I, Hind III and ligation, PRMT1 was inserted into pcDNA3.1(+)eukaryotic expression vector. The positive colonies were screened and identified by PCR and sequencing. pcDNA3.1(+)-PRMT1 plasmid was then transfected into the cultured A549 cell line with Lipofectamine(TM);2000. Realtime-PCR and Western blot were used to detect the mRNA and protein expression of PRMT1 respectively. RESULTS: The PRMT1 cDNA was successfully amplified, and pcDNA3.1(+)-PRMT1 were constructed. The inserted sequence in pcDNA3.1(+)-PRMT1 was the same as the sequence of PRMT1 cDNA published in NCBI GenBank. Further, Realtime PCR and Western blot results validated the recombinant plasmid expressed in A549 cell line efficiently. CONCLUSION: pcDNA3.1(+)-PRMT1 recombinant was successfully constructed.

Effects of extracellular Ca(2+) influx and intracellular Ca(2+) release on ethanol-induced cytoplasmic Ca(2+) overload in cultured superior cervical ganglion neurons.

The present research was designed to investigate the interference of Ca(2+) homeostasis by ethanol on the primary cultured superior cervical ganglion (SCG) neurons. (1) Using the whole cell patch clamp recording, the amplitudes of voltage-dependent Ca(2+) channel (VDCC) currents could be reduced by ethanol in a concentration-dependent manner. Ethanol (100mM) inhibited about 25% of Ca(2+) channel current. However, the activation of Ca(2+) channel was not affected by ethanol at those concentrations. (2) The similar extent inhibitions of 100mM ethanol on the increments of intracellular Ca(2+) concentration ([Ca(2+)](i)) induced by 40 mM KCl and 1 microM A23187 were also observed in the fluo-3-AM loaded superior cervical ganglia (SCG) via detecting the change of [Ca(2+)](i) with a laser scanning confocal microscopy. In contrast, the basal [Ca(2+)](i) was significantly increased by ethanol alone in a concentration-dependent manner. These phenomena were also observed even under Ca(2+) free bath solution or the solution added 300 microM cadmium chloride conditions. Together with above results, our data suggest that ethanol increases basal [Ca(2+)](i), but it also inhibits the extracellular Ca(2+) influx through VDCC and ionophore channel. And the augment of basal [Ca(2+)](i) induced by ethanol might attribute to the Ca(2+) releasing from intracellular Ca(2+) pools.

[Effects of iodophor on the thyroid glands of female medical workers].

OBJECTIVE: To investigate the long-term effects of iodophor on thyroid function and autoimmunity in females. METHODS: 119 medical personnel who have been using iodophor as preoperative skin disinfectant more for than 2 years and 123 medical personnel who have not exposed to iodophor were studied. The urinary iodine, thyroid-stimulating hormone (TSH), free triiodothyronine (FT(3)), free thyroxine (FT(4)), thyroid peroxydase antibody (TPO-Ab) and TG-Ab in serum were measured and the thyroid were examined by B mode ultrasound apparatus. RESULTS: The median urinary iodine (MUI) was 300.4 micro g/L in the exposed group, not significantly higher than that in the non-exposed group (269.1 micro g/L, P > 0.05). The TSH levels of the exposed group and non-exposed group were 1.66 mU/L and 1.62 mU/L (P = 0.84). B mode ultrasound examination showed that the prevalence rate of thyroid disorders was 3.36% in the exposed group, not significantly different from that in the non-exposed group (2.44%). The thyroid autoantibody positive rate was not significantly different between these two groups too. The titer of autoantibody in the exposed group was 29.5 IU/L, significantly higher than that in the non-exposed group (22.4 IU/L, P = 0.048). The titer of TG-Ab in the exposed group was 21.85 IU/L, not significantly different from that in the non-exposed group (18.7 IU/L, P = 0.542). The mean titer of TPO-Ab in exposed group was 29.5 IU/L, significantly higher than that in non-exposed group (22.4 IU/L, P = 0.048). The total prevalence rate of all sorts of thyroid disorders in the exposed group was 11.76%, significantly higher than that in non-exposed group (4.07%, P = 0.026). The thyroid disorders discovered included clinical hyperthyroidism and subclinical hypothyroidism. CONCLUSION: Using iodophor as preoperative skin disinfectant may result in an increased incidence of thyroid disorders in female medical personnel. Medical personnel with iodophor exposure history should have their thyroid function and thyroid autoimmune status examined regularly.