Stevioside attenuates bleomycin-induced pulmonary fibrosis by activating the Nrf2 pathway and inhibiting the NF-κB and TGF-ß1/Smad2/3 pathways.

Objective: This study aimed to investigate the effects of stevioside (STE) on pulmonary fibrosis (PF) and the potential mechanisms. Methods: In this study, a mouse model of PF was established by a single intratracheal injection of bleomycin (BLM, 3 mg/kg). The experiment consisted of four groups: control group, BLM group, and STE treatment groups (STE 50 and 100 mg/kg). ELISA and biochemical tests were conducted to determine the levels of TNF-α, IL-1ß, IL-6, NO, hydroxyproline (HYP), SOD, GSH, and MDA. Histopathological changes and collagen deposition in lung tissues were observed by HE and Masson staining. Immunohistochemistry was performed to determine the levels of collagen I-, collagen III-, TGF-ß1- and p-Smad2/3-positive cells. Western blot analysis was used to measure the expression of epithelial-mesenchymal transition (EMT) markers, including α-SMA, vimentin, E-cadherin, and ZO-1, as well as proteins related to the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, nuclear transcription factor-κB (NF-κB) pathway, and TGF-ß1/Smad2/3 pathway in lung tissues. Results: STE significantly alleviated BLM-induced body weight loss and lung injury in mice, decreased HYP levels, and reduced the levels of collagen I- and collagen III-positive cells, thereby decreasing extracellular matrix (ECM) deposition. Moreover, STE markedly improved oxidative stress (MDA levels were decreased, while SOD and GSH activity were enhanced), the inflammatory response (the levels of TNF-α, IL-1ß, IL-6, and NO were reduced), and EMT (the expression of α-SMA and vimentin was downregulated, and the expression of E-cadherin and ZO-1 was upregulated). Further mechanistic analysis revealed that STE could activate the Nrf2 pathway and inhibit the NF-κB and TGF-ß1/Smad2/3 pathways. Conclusion: STE may alleviate oxidative stress by activating the Nrf2 pathway, suppress the inflammatory response by downregulating the NF-κB pathway, and inhibit EMT progression by blocking the TGF-ß1/Smad2/3 pathway, thereby improving BLM-induced PF.

Analytical Method Optimization of Tetrodotoxin and Its Contamination in Gastropods.

Tetrodotoxin (TTX) is an extremely potent marine biotoxin. An analytical method was developed for both trace contamination and extremely high levels of TTX in gastropods by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with clean-up of cation exchange solid phase extraction (SPE) in this study. The limit of detection (LOD) in the sample matrix was 0.5 µg/kg. With the calibration of a screened internal standard (validamycin, IS), the linear range was 0.1-100 ng/mL (1.5-1500 µg/kg in sample matrix) with a correlation coefficient of r2 > 0.999. The average recoveries at three spiking levels (1.5 µg/kg, 44 µg/kg, and 1500 µg/kg) were 82.6-94.4% with relative standard deviations (RSDs) less than 8.4%. TTX levels in seven gastropods (741 samples) were studied. The contamination and analogues in Neverita didyma (N. didyma, 565 samples collected in Zhejiang province, China, from 2016 to 2022) were first reported. The detection rate of TTX in N. didyma was 34.2%. The average concentration was 23.1 µg/kg, and the maximum value was 2327 µg/kg. The time distribution study indicated that high contaminations of TTX occurred from May to August for N. didyma.

CEUS LI-RADS for diagnosis of hepatocellular carcinoma in individuals without LI-RADS-defined hepatocellular carcinoma risk factors.

PURPOSE: This study evaluated the performance of the contrast-enhanced ultrasound (CEUS) Liver Imaging Reporting and Data System (LI-RADS) in patients without LI-RADS-defined hepatocellular carcinoma (HCC) risk factors (RF-). METHODS: Patients with LI-RADS-defined HCC risk factors (RF+) and RF- were enrolled in a retrospective study. Additionally, a prospective evaluation in the same centre was performed as a validation set. The diagnostic performances of the CEUS LI-RADS criteria in RF+ and RF- patients were compared. RESULTS: Overall, we included 873 patients in the analyses. In the retrospective study, the LI-RADS category (LR)-5 specificities for diagnosing HCC did not differ between the RF+ and RF- groups (77.5% [158/204] vs 91.6% [196/214], P = 0.369, respectively). However, the positive predictive value (PPV) of CEUS LR-5 was 95.9% (162/169) and 89.8% (158/176) in the RF+ and RF- groups, respectively (P = 0.029). In the prospective study, the PPV of LR-5 for HCC lesions was significantly higher in the RF+ group than in the RF- group (P = 0.030). The sensitivity and specificity did not differ between the RF+ and RF- groups (P = 0.845 and P = 0.577, respectively). CONCLUSIONS: The CEUS LR-5 criteria shows clinical value for diagnosis of HCC in patients with and without risks.

Mechanisms of Chinese Medicine in Gastroesophageal Reflux Disease Treatment: Data Mining and Systematic Pharmacology Study.

OBJECTIVE: To identify specific Chinese medicines (CMs) that may benefit patients with gastroesophageal reflux disease (GERD), and explore the action mechanism. METHODS: Domestic and foreign literature on the treatment of GERD with CMs was searched and selected from China National Knowledge Infrastructure, China Science and Technology Journal Database, Wanfang Database, and PubMed from October 1, 2011 to October 1, 2021. Data from all eligible articles were extracted to establish the database of CMs for GERD. Apriori algorithm of data mining techniques was used to analyze the rules of herbs selection and core Chinese medicine formulas were identified. A system pharmacology approach was used to explore the action mechanism of these medicines. RESULTS: A total of 278 prescriptions for GERD were analyzed, including 192 CMs. Results of Apriori algorithm indicated that Evodiae Fructus and Coptidis Rhizoma were the highest confidence combination. A total of 32 active ingredients and 66 targets were screened for the treatment of GERD. Enrichment analysis showed that the mechanisms of action mainly involved pathways in cancer, fluid shear stress and atherosclerosis, advanced glycation end product (AGE), the receptor for AGE signaling pathway in diabetic complications, bladder cancer, and rheumatoid arthritis. CONCLUSION: Evodiae Fructus and Coptidis Rhizoma are the core drugs in the treatment of GERD and the potential mechanism of action of these medicines includes potential target and pathways.

Analysis of furan and its major furan derivatives in coffee products on the Chinese market using HS-GC-MS and the estimated exposure of the Chinese population.

The current study analysed concentrations of furan and its derivatives in coffee products commercially available in China based on an improved headspace gas chromatography-mass spectrometry (HS-GC-MS) method and estimated health risks. A total of 101 samples of coffee products on the Chinese market was analysed. Furan (98%, ND-6569 µg/kg) and 2-methylfuran (100%, 2-29639 µg/kg) were the compounds with the highest concentrations and detection rates in coffee products. The mean dietary exposure of Chinese consumers to furan and the sum of furan, 2-methylfuran and 3-methylfuran in coffee products was 0.09 and 0.46 µg/kg bw/day, respectively. For the neoplastic effects of furan, the margin of exposure (MOE) was 14,556 for the mean dietary exposure of consumers. For the non-neoplastic effects of furan, MOEs were 711 and 139 for furan and the sum of furan, 2-methylfuran and 3-methylfuran, respectively. Overall, a health concern is indicated for coffee consumers with MOEs below 10000.

Cellulosic hydrocarbons production by engineering dual synthesis pathways in Corynebacterium glutamicum.

BACKGROUND: Lignocellulose provides the only practical carbohydrates feedstock for sustainable bioproduction of hydrocarbons as future alternative of fossil fuels. Production of hydrocarbons from lignocellulose is achieved by a biorefinery process chain including pretreatment to breakdown the crystalline structure for cellulase-catalyzed hydrolysis, detoxification of inhibitory compounds generated during pretreatment, enzymatic hydrolysis to fermentable monosaccharide sugars, and fermentation to hydrocarbon products. The major barriers on fermentative production of hydrocarbons from lignocellulose include two aspects: one is the inherent stress of pretreatment-derived inhibitors on microbial cells, the other is the toxicity of hydrocarbons to cell membranes. The microbial cell factory should be tolerant to both inhibitor stress and hydrocarbons toxicity. RESULTS: Corynebacterium glutamicum was selected as the starting strain of hydrocarbons synthesis since it is well adapted to lignocellulose hydrolysate environment. The dual hydrocarbon synthesis pathways were constructed in an industrial C. glutamicum S9114 strain. The first pathway was the regular one in microalgae composed of fatty acyl-acyl carrier protein (fatty acyl-ACP) reductase (AAR) and aldehyde deformylating oxygenase (ADO) with fatty acyl-ACP as precursor. The second pathway was the direct decarboxylation of free fatty acid by fatty acid decarboxylase (OleT) using the rich fatty acids from the disruption of the transcriptional regulator fasR gene. The transmembrane transportation of hydrocarbon products was avoided by secretively expressing the fatty acid decarboxylase (OleT) to the extracellular space. The hydrocarbons generation from glucose reached 29.2 mg/L, in which the direct decarboxylation pathway contributed more than 70% of the total hydrocarbons generation, and the AAR-ADO pathway contributed the rest 30%. CONCLUSION: The dual hydrocarbon synthesis pathways (OleT and AAR-ADO pathways) were constructed in the inhibitors tolerant C. glutamicum S9114 strain for hydrocarbon production using lignocellulose feedstock as the starting feedstock. When corn stover was used for hydrocarbons production after dry acid pretreatment and biodetoxification, the hydrocarbons generation reached 16.0 mg/L. This study provided a new strategy for hydrocarbons synthesis using microbial cell factory suitable for lignocellulose feedstock.

Genomic characterization of a new CRF01_AE/CRF07_BC case from a MSM patient in Guangdong, China.

The CRF01_AE and CRF07_BC clades dominate the human immunodeficiency virus (HIV) epidemics in China. Both clades have been identified in the men who have sex with men (MSM) population in Guangdong province, raising a serious concern of possible complex recombination events ahead. Here, we report the first case of CRF01_AE/CRF07_BC recombinant sampled from a MSM patient in southern China. The genomic structure of this case is a mosaic with some regions resembling the CRF01_AE and CRF07_BC clades. Our phylogenetic analyses show that the two parental lineages of this recombinant virus were mainly found in the MSM population. This case has a different genomic composition compared with other recombinants descended from the same parental clades CRF01_AE and CRF07_BC. Our finding suggests that the MSM populations have become a hotspot for expanding viral diversity through the viral recombination mechanism. Therefore, further epidemiologic surveillance and monitoring should be conducted within the MSM populations to help advance our knowledge of viral transmission mechanisms. Additionally, these measures will serve to enhance the control and prevention of HIV/acquired immunodeficiency syndrome in China.

Rapid Development of SARS-CoV-2 Spike Protein Receptor-Binding Domain Self-Assembled Nanoparticle Vaccine Candidates.

The coronavirus disease pandemic of 2019 (COVID-19) caused by the novel SARS-CoV-2 coronavirus resulted in economic losses and threatened human health worldwide. The pandemic highlights an urgent need for a stable, easily produced, and effective vaccine. SARS-CoV-2 uses the spike protein receptor-binding domain (RBD) to bind its cognate receptor, angiotensin-converting enzyme 2 (ACE2), and initiate membrane fusion. Thus, the RBD is an ideal target for vaccine development. In this study, we designed three different RBD-conjugated nanoparticle vaccine candidates, namely, RBD-Ferritin (24-mer), RBD-mi3 (60-mer), and RBD-I53-50 (120-mer), via covalent conjugation using the SpyTag-SpyCatcher system. When mice were immunized with the RBD-conjugated nanoparticles (NPs) in conjunction with the AddaVax or Sigma Adjuvant System, the resulting antisera exhibited 8- to 120-fold greater neutralizing activity against both a pseudovirus and the authentic virus than those of mice immunized with monomeric RBD. Most importantly, sera from mice immunized with RBD-conjugated NPs more efficiently blocked the binding of RBD to ACE2 in vitro, further corroborating the promising immunization effect. Additionally, the vaccine has distinct advantages in terms of a relatively simple scale-up and flexible assembly. These results illustrate that the SARS-CoV-2 RBD-conjugated nanoparticles developed in this study are a competitive vaccine candidate and that the carrier nanoparticles could be adopted as a universal platform for a future vaccine development.

Kinetic Characteristics of Neutralizing Antibody Responses Vary among Patients with COVID-19.

OBJECTIVE: The coronavirus disease 2019 (COVID-19) pandemic continues to present a major challenge to public health. Vaccine development requires an understanding of the kinetics of neutralizing antibody (NAb) responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). METHODS: In total, 605 serum samples from 125 COVID-19 patients (from January 1 to March 14, 2020) varying in age, sex, severity of symptoms, and presence of underlying diseases were collected, and antibody titers were measured using a micro-neutralization assay with wild-type SARS-CoV-2. RESULTS: NAbs were detectable approximately 10 days post-onset (dpo) of symptoms and peaked at approximately 20 dpo. The NAb levels were slightly higher in young males and severe cases, while no significant difference was observed for the other classifications. In follow-up cases, the NAb titer had increased or stabilized in 18 cases, whereas it had decreased in 26 cases, and in one case NAbs were undetectable at the end of our observation. Although a decreasing trend in NAb titer was observed in many cases, the NAb level was generally still protective. CONCLUSION: We demonstrated that NAb levels vary among all categories of COVID-19 patients. Long-term studies are needed to determine the longevity and protective efficiency of NAbs induced by SARS-CoV-2.

Efficacy of transconjunctival approach for the treatment of orbital fractures: A protocol for systematic review and meta-analysis.

BACKGROUND: This study aims to assess the efficacy of transconjunctival approach (TCA) for the treatment of orbital fractures (OF) comprehensively and systematically. METHODS: In this study, we plan to search electronic databases of Cochrane Library, MEDLINE, EMBASE, Web of Science, Allied and Complementary Medicine Database, Chinese Biomedical Literature Database, China National Knowledge Infrastructure and for relevant randomized controlled trials. All these databases will be searched from inception to the March 1, 2020 without limitations of language and publication status. Two independent authors will carry out study selection, data collection, and study quality assessment. Any disagreements will be resolved by discussion with another author if necessary. The study quality will be assessed using Cochrane risk of bias tool. Statistical analysis will be conducted using RevMan 5.3 software. RESULTS: This study will be the first 1 to exert direct evidence to evaluate the efficacy of TCA for the treatment of OF. CONCLUSIONS: The findings of this study will provide an exhaustive view of TCA for the treatment of OF. STUDY REGISTRATION NUMBER: INPLASY202040154.

Canaloplasty for the treatment of primary open-angle glaucoma: A protocol of systematic review and meta-analysis.

BACKGROUND: Canaloplasty has been reported to manage primary open-angle glaucoma (POAG) effectively. However, no study has specifically and systematically investigated the efficacy and safety of canaloplasty for the treatment of POAG. Thus, this study will systematically and comprehensively appraise the efficacy and safety of canaloplasty for the treatment of POAG. METHODS: MEDLINE, EMBASE, Cochrane Library, Web of Science, Cumulative Index to Nursing and Allied Health Literature, Allied and Complementary Medicine Database, Chinese Biomedical Literature Database, and China National Knowledge Infrastructure will be sought from the construction to the February 29, 2020. Only randomized controlled trials (RCTs) focusing on canaloplasty for the treatment of POAG will be included. Two reviewers will independently undertake selection of study, data extraction, and risk of bias assessment. Any doubts between 2 reviewers will be resolved through discussion with another experienced reviewer. RevMan 5.3 software will be employed for data analysis. RESULTS: This study will summarize high-quality RCTs on investigating efficacy and safety of canaloplasty for the treatment of POAG. CONCLUSION: The findings of this study will help to determine whether canaloplasty is effective and safety for the treatment of POAG.Systematic review registration: INPLASY202040119.

Acupuncture treatment for optic atrophy: A protocol for systematic review.

BACKGROUND: Optic atrophy (OPA) is a very tricky disorder. Presently, no effective management is available for this condition. Previous studies have reported that acupuncture may be effective for the treatment of OPA. However, its effectiveness is still inconclusive. Thus, this study will aim to assess the effectiveness and safety of acupuncture for OPA. METHODS: A comprehensive literature search for relevant studies will be performed from the databases of PUMBED, EMBASE, CINAHI, Cumulative Index to Nursing and Allied Health Literature, Allied and Complementary Medicine Database, Cochrane Library, Chinese Biomedical Literature Database, China National Knowledge Infrastructure, and other literature sources from inception up to the present. No language limitations will be applied to all literature searches. We will consider all randomized controlled trials (RCTs) and case-controlled trials (CCTs) for assessing the effectiveness and safety of acupuncture for OPA. The primary outcomes include the rates of vision improvement and visual field improvement. The secondary outcomes consist of the increased visual field average sensitivity, pattern visual evoked potential (PVEP) amplitude, and shortened PVEP latency, as well as any expected and unexpected adverse reactions. Risk of bias assessment will be performed by Cochrane risk of bias for RCTs and Newcastle-Ottawa Scale for CCTs. RESULTS: In this study, we will outline details of the aims and methods on the effectiveness and safety of acupuncture for the treatment of OPA. CONCLUSION: The results of this study will summarize the most current evidence of acupuncture for the treatment of patients with OPA. DISSEMINATION AND ETHICS: The results of this study are expected to be published on peer-reviewed journals. This is a literature-based study; therefore, no ethical approval is necessary. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42019135785.

Complicated HCV subtype expansion among drug users in Guangdong province, China.

Guangdong Province is one of the most developed and populous provinces in southern China. The subtype situation of hepatitis C virus (HCV) in Guangdong remains unknown. The aim of this study was to investigate and estimate the HCV subtypes in drug users (DU) using a city-based sampling strategy to better understand the characteristics of HCV transmission in Guangdong. Archived plasma samples (n = 1074) from DU who were anti-HCV positive in 2014 were selected randomly from 20 cities in Guangdong Province. Subtypes were determined based on core and/or E1 sequences using phylogenetic analysis. The distributions of HCV subtypes in DU and different regions were analyzed. A total of 8 genotypes were identified. The three main HCV subtypes in DU in Guangdong were 6a (63.0%), 3a (15.2%), and 3b (11.8%). Significant differences were discovered among different registered residency and regions but not among genders, marital status, education level, or drug use patterns. HCV subtype 3b was significantly higher in Guangdong residents than in non-Guangdong residents. In contrast, HCV subtype 6a was significantly lower in Guangdong residents than in non-Guangdong residents. Subtype 1b in eastern Guangdong (eastern) was significantly lower, while 6a was significantly higher when compared with other regions. Subtype 3a in the Pearl River Delta (PRD) region was significantly higher, while 3b was significantly lower when compared with other regions. In western Guangdong, HCV subtype 3a was significantly lower when compared with other regions. Additionally, in northern Guangdong subtypes 1b and 3b were significantly higher, while 6a was significantly lower when compared with other regions. Our study revealed the diversity and distribution of HCV subtypes in DU in nearly all the cities in Guangdong. The results provide essential information that will allow the establishment of specific intervention strategies that may help prevent HCV transmission.

Omega-3 and ranibizumab for age-related macular degeneration: A systematic review protocol.

BACKGROUND: Omega-3 and ranibizumab (O3R) has been reported to treat age-related macular degeneration (ARMD) effectively. However, up to the present, no systematic review specifically addressed the efficacy of O3R for the treatment of ARMD. Therefore, in this study, we will propose to assess the efficacy and safety of O3R for the treatment of ARMD. METHODS: We will search PUMBED, EMBASE, CINAHI, Cumulative Index to Nursing and Allied Health Literature, Allied and Complementary Medicine Database, Cochrane Library, Chinese Biomedical Literature Database, China National Knowledge Infrastructure, VIP Information, Wanfang Data, as well as the gray literature from inception up to the present. We will accept randomized controlled trials for assessing the efficacy and safety of O3R for ARMD. The primary outcomes include change in best corrected visual acuity and central retinal thickness. The secondary outcomes consist of changes in subfoveal choroidal thickness, macular atrophy, retinal average sensitivity, contrast sensitivity, glare disability, and quality of life. In addition, incidence and severity of adverse events will also be evaluated. Cochrane Collaboration tool will be used to assess the risk of bias for each included study. In addition, Grading of Recommendations Assessment, Development, and Evaluation tool will be utilized to assess the overall strength of the evidence. Two authors will independently carry out all procedures and any divergences will be solved through discussion with a third author. If it is possible, we will conduct meta-analysis and subgroup analysis concerning different interventions, risk of bias, and outcome measurements. RESULTS: In this proposed study, we outline details of the aims and methods of efficacy and safety of O3R for the treatment of ARMD. CONCLUSION: The findings of this systematic review will summarize current evidence of O3R for the treatment of patients with ARMD. DISSEMINATION AND ETHICS: The results of the present study are expected to be published by peer-reviewed journals. This is a literature-based study. Thus, ethical approval is unnecessary for this study. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42019121177.

Rapid and complicated HIV genotype expansion among high-risk groups in Guangdong Province, China.

BACKGROUND: Guangdong Province is one of the most developed and populous provinces in southern China, with frequent foreign exchanges and large transient population. The annual number of cases of HIV/AIDS reported in Guangdong has been higher than most of provinces in China for several successive years. HIV infection by heterosexual transmission occurs across the province, with transmission among men who have sex with men occurring mainly in larger urban centers. There is a lack of widespread and representative data on the distribution of HIV subtypes in Guangdong. This study aimed to thoroughly investigate and estimate the prevalence and distribution of HIV-1 subtypes using a city-based sampling strategy to better understand the characteristics of HIV transmission in Guangdong. METHODS: Archived plasma samples (n = 1205) from individuals diagnosed as HIV-1 infection in 2013 were selected randomly from all 21 cities in Guangdong Province. Genotypes were determined using env and/or gag sequences using phylogenetic analysis. The distributions of HIV genotypes in different risk groups and different cities were analyzed. RESULTS: A total of 15 genotypes, including six discordant genotypes, were identified. The four main HIV-1 subtypes in Guangdong were CRF01_AE (43.2%), CRF07_BC (26.3%), CRF55_01B (8.5%), and CRF08_BC (8.4%). CRF01_AE was the predominant subtype in all risk populations. The high mobility of people shaped the complexity of the HIV genotypes, while the switch of risk factors affected the distribution and future trend of HIV-1 genotypes in Guangdong. Another epicenter located in the western region in addition to the known epicenter cities in the Pearl River Delta region of Guangdong may exist. CONCLUSIONS: Our study provides a comprehensive molecular epidemiologic dataset to understand the diversity and distribution of HIV genotypes in Guangdong, as well as to clarify the unique region- and risk group-specific transmission dynamics. The results provide critical and insightful information for more effective intervention strategies to limit HIV transmission in the future.

Knockdown of hnRNP A2/B1 inhibits cell proliferation, invasion and cell cycle triggering apoptosis in cervical cancer via PI3K/AKT signaling pathway.

Cervical cancer is currently one of the major threats to women's health. The overexpression of heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1) as the biomarker has been investigated in various cancers. In our previous study, we found that lobaplatin induced apoptosis and cell cycle arrest via downregulation of proteins including hnRNP A2/B1 in cervical cancer cells. However, the underlying relationship between hnRNP A2/B1 and cervical cancer remained largely unknown. hnRNP A2/B1 knock­down in HeLa and CaSki cells was performed by shRNA transfection. The expression of hnRNP A2/B1 was detected by western blot and Quantitative Real­time PCR. Cell proliferation, migration, invasion and the IC50 of lobaplatin and irinotecan were determined by MTT assay, Transwell assay, Plate colony formation assay and wound healing assay. Flow cytometry was perfomed to investigate cell apoptosis and the cell cycle. The expression of PI3K, AKT, p­AKT, p21, p27, caspase­3, cleaved caspase­3 were revealed by western blot. Nude mouse xenograft model was undertaken with HeLa cells and the xenograft tumor tissue samples were analyzed for the expression of PCNA and Ki­67 by immunohistochemistry and the cell morphology was evaluated by hematoxylin and eosin (H&E). Results revealed that hnRNP A2/B1 was successfully silenced in HeLa and CaSki cells. hnRNP A2/B1 knock­down significantly induced the suppression of proliferation, migration, invasion and also enhancement of apoptosis and reduced the IC50 of lobaplatin and irinotecan. The expression of p21, p27 and cleaved caspase­3 in shRNA group were significantly upregulated and the expression of p­AKT was reduced both in vitro and in vivo. The results of immunohistochemistry showed that PCNA and Ki­67 were significantly downregulated in vivo. The growth of nude mouse xenograft tumor was significantly reduced by hnRNP A2/B1 knock­down. Taken together, these data indicate that inhibition of hnRNP A2/B1 in cervical cancer cells can inhibit cell proliferation and invasion, induce cell­cycle arrestment and trigger apoptosis via PI3K/AKT signaling pathway. In addition, after silencing hnRNP A2/B1 can increase the sensitivity of cervical cancer cells to lobaplatin and irinotecan.

Fermentative production of high titer citric acid from corn stover feedstock after dry dilute acid pretreatment and biodetoxification.

The aim of this work is to study the citric acid fermentation by a robust strain Aspergillus niger SIIM M288 using corn stover feedstock after dry dilute sulfuric acid pretreatment and biodetoxification. Citric acid at 100.04g/L with the yield of 94.11% was obtained, which are comparable to the starch or sucrose based citric acid fermentation. No free wastewater was generated in the overall process from the pretreatment to citric acid fermentation. Abundant divalent metal ions as well as high titer of potassium, phosphate, and nitrogen were found in corn stover hydrolysate. Further addition of extra nutrients showed no impact on increasing citric acid formation except minimum nitrogen source was required. Various fermentation parameters were tested and only minimum regulation was required during the fermentation. This study provided a biorefining process for citric acid fermentation from lignocellulose feedstock with the maximum citric acid titer and yield.

[Role of changes in sodium pump activity and endoplasmic reticulum stress in the ischemia/reperfusion induced injury of isolated hearts].

OBJECTIVE: To investigate the roles of change in sodium pump activity and endoplasmic reticulum stress (ERS) in ischemia/reperfusion (I/R) injury of the isolated rat hearts. METHODS: Sixty male SD rats were randomly divided into 6 groups (n=10 each):normal control group (NC), I/R group (I/R), ouabain-I/R group (OUA-I/R), anti-digoxin antiserum-I/R group (Anti-Dig-I/R), PP2 (Src kinase inhibitor)-ouabain-I/R group (PP2-OUA-I/R),U73122 (PLC inhibitor)-ouabain-I/R group (PP2-OUA-I/R). The isolated rat hearts were perfused on the Langendorff apparatus. Except for NC group, all the hearts were subjected to 30 min global ischemia and followed by 120 min reperfusion. The cardiac function indexes were recorded at the same time. The coronary effluent was collected for estimating the activity of lactate dehydrogenase (LDH) and creatine kinase (CK). The activity of Na+-K+-ATPase and intracellular calcium concentration in myocardial tissue were measured. Apoptosis was evaluated by Flow cytometric analysis. The expressions of sodium pump α1 subunit, glucose regulated protein(GRP78),C/EBP homologous protein (CHOP) and Bcl-2/Bax were determined by Western blot. RESULTS: Pretreatment with ouabain significantly reduced the recovery of cardiac function, increased the levels of CK, LDH and intracellular calcium concentration, decreased the activity of Na+-K+-ATPase. In addition, ouabain markedly increased the myocardial apoptosis index, down-regulated the expressions of sodium pump α1 subunit and Bcl-2, up-regulated the expressions of GRP78,CHOP and Bax; while these changes were significantly improved in the Anti-Dig-I/R group compared with those in the I/R group; PP2 or U73122 partially blocked the effects of ouabain on myocardial I/R injury. Compared with the OUA-IR group, the recovery of cardiac function, the activity of Na+-K+-ATPase and the expressions of sodium pump α1 subunit and Bcl-2 were significant higher, meanwhile the leakage of CK and LDH, intracellular calcium concentration, myocardial apoptosis index and the expressions of GRP78 and Bax were significantly lower in PP2-OUA-I/R and U73122-OUA-I/R group. CONCLUSIONS: Changes in sodium pump function and endoplasmic reticulum stress all participate in the process of I/R injury. Current findings further suggest that sodium pump mediates ERS by activating signals of Src and PLC pathway, which may be one of the mechanisms of apoptosis induced by I/R injury.

Heme Oxygenase-1 Promotes Delayed Wound Healing in Diabetic Rats.

Diabetic ulcers are one of the most serious and costly chronic complications for diabetic patients. Hyperglycemia-induced oxidative stress may play an important role in diabetes and its complications. The aim of the study was to explore the effect of heme oxygenase-1 on wound closure in diabetic rats. Diabetic wound model was prepared by making an incision with full thickness in STZ-induced diabetic rats. Wounds from diabetic rats were treated with 10% hemin ointment for 21 days. Increase of HO-1 protein expression enhanced anti-inflammation and antioxidant in diabetic rats. Furthermore, HO-1 increased the levels of VEGF and ICAM-1 and expressions of CBS and CSE protein. In summary, HO-1 promoted the wound closure by augmenting anti-inflammation, antioxidant, and angiogenesis in diabetic rats.

[Effects of total flavonoids of propolis on apoptosis of myocardial cells of chronic heart failure and its possible mechanism in rats].

OBJECTIVE: To investigate the effects of total flavonoids of propolis (TFP) on apoptosis of myocardial cells of chronic heart failure and its possible mechanism in rats. METHODS: Six male SD rats were randomly selected as normal control group, the remaining rats were made as chronic heart failure (CHF) model by intraperitoneal injection of adriamycin. The rats in the successful model were randomly divided into five groups (n = 6): CHF group, total flavonoids of propolis low dose group (LD group), total flavonoids of propolis middle dose group (MD group), total flavonoids of propolis high dose group (HD group), digoxin group (DIG group). After six week treatment, cardiac function indexes of rats were recorded by signal acquisition system; brain natriuretic peptide (BNP), cardiac troponin I (cTnI), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) content in plasma were detected; Myocardial morphological changes and collagen fiber hyperplasia by HE and Masson staining were observed; Myocardial apoptosis was detected with TUNEL method and protein connexin 43(P-Cx43) expression was detected by Western blot method. RESULTS: Compared with NC group, left ventricular systolic pressure(LVSP) and maximal rise/fall velocity of left ventriculad pressure (± dP/dt(max)) absolute value in CHF group were significantly lowered (P < 0.01) while left ventricular end diastolic pressure (LVEDP) was increased significantly (P < 0.01); Contents of plasma BNP, cTnI, TNF-α and IL-6 in the CHF group were significantly improved (P < 0.01). Compared with CHF group, LVSP, ± dP/dt(max) absolute value in MD and HD groups were increased (P < 0.05), and LVEDP was significantly lowered (P < 0.01); LVEDP in LD group was significantly lowered (P < 0.01), changes in LVSP and ± dp/dt(max) absolue value were not obvious (P > 0.05). BNP, cTnI, TNF-α and IL-6 contents in MD and HD groups were significantly reduced (P < 0.01), but those plasma indicator changes were not obvious in LD group (P > 0.05). Western blot showed that P-Cx43 expression in CHF group was significantly higher than that in NC group (P < 0.01) and that in all TFP treatment groups it was decreased compared with CHF group (P < 0.05, P < 0.01), among which pairwise comparisons also showed differences (P < 0.05), myocardial apoptosis index (%)(22.62 ± 3.39) in CHF group was higher than that in NC group( 1.12 ± 0.24) (P < 0.01); compared with CHF group, the apoptosis index of myocardial cells (%) in LD,MD and HD groups, (15.79 + 2.8), (9.28 + 2.1) and (4.73 + 1.14) respectively, were significantly lower than those in the CHF group( P < 0.01). The expression level of P-Cx43 positively correlated with the apoptotic index (r = 0. 861, P < 0.01). CONCLUSION: Total flavonaids of propolis have inhibitory effect on apoptosis of myocardial cells of chronic heart failure induced by adriamycin in rats, and the mechanism may be closely related to the regulation of Cx43 expression, especially the regulatory phosphorylation status.