Electroacupuncture Improves Cardiac Function via Inhibiting Sympathetic Remodeling Mediated by Promoting Macrophage M2 Polarization in Myocardial Infarction Mice.

Electroacupuncture (EA) at the Neiguan acupoint (PC6) has shown significant cardioprotective effects. Sympathetic nerves play an important role in maintaining cardiac function after myocardial infarction (MI). Previous studies have found that EA treatment may improve cardiac function by modulating sympathetic remodeling after MI. However, the mechanism in how EA affects sympathetic remodeling and improves cardiac function remains unclear. The aim of this study is to investigate the cardioprotective mechanism of EA after myocardial ischemic injury by improving sympathetic remodeling and promoting macrophage M2 polarization. We established a mouse model of MI by occluding coronary arteries in male C57/BL6 mice. EA treatment was performed at the PC6 with current intensity (1 mA) and frequency (2/15 Hz). Cardiac function was evaluated using echocardiography. Heart rate variability in mice was assessed via standard electrocardiography. Myocardial fibrosis was evaluated by Sirius red staining. Levels of inflammatory factors were assessed using RT-qPCR. Sympathetic nerve remodeling was assessed through ELISA, western blotting, immunohistochemistry, and immunofluorescence staining. Macrophage polarization was evaluated using flow cytometry. Our results indicated that cardiac systolic function improved significantly after EA treatment, with an increase in fractional shortening and ejection fraction. Myocardial fibrosis was significantly mitigated in the EA group. The sympathetic nerve marker tyrosine hydroxylase and the nerve sprouting marker growth-associated Protein 43 were significantly reduced in the EA group, indicating that sympathetic remodeling was significantly reduced. EA treatment also promoted macrophage M2 polarization, reduced levels of inflammatory factors TNF-α, IL-1ß, and IL-6, and decreased macrophage-associated nerve growth factor in myocardial tissue. To sum up, our results suggest that EA at PC6 attenuates sympathetic remodeling after MI to promote macrophage M2 polarization and improve cardiac function.

Fluorine-free preparation of a superhydrophobic coating with anti-icing properties, mechanical durability and self-cleaning effect.

Superhydrophobic materials have become a feasible choice to solve related difficult problems because of their excellent anti-icing, anti-corrosion, and self-cleaning characteristics. In this work, a superhydrophobic hydroxypropyl methylcellulose (HPMC)/SiO2 coating is prepared using an efficient, fluorine-free method for the anti-icing application of transmission line insulators and other similar material surfaces. The water contact angle (WCA) of the coating is 161°, and the slide angle (SA) is less than 1°. The coating maintains good hydrophobicity after mechanical durability tests. In the anti-icing performance tests, the start freezing time of a single droplet is delayed by 1366 s, and when the surface is not coated, the ice amount is more than twice that with the coating. Therefore, this work provides a straightforward and promising solution to solving high-cost and low-efficiency difficulties in the anti-icing problem of transmission line insulators and other similar material surfaces.

Purinergic signaling in myocardial ischemia-reperfusion injury.

Purines and their derivatives, extensively distributed in the body, act as a class of extracellular signaling molecules via a rich array of receptors, also known as purinoceptors (P1, P2X, and P2Y). They mediate multiple intracellular signal transduction pathways and participate in various physiological and pathological cell behaviors. Since the function in myocardial ischemia-reperfusion injury (MIRI), this review summarized the involvement of purinergic signal transduction in diversified pathological processes, including energy metabolism disorder, oxidative stress injury, calcium overload, inflammatory immune response, platelet aggregation, coronary vascular dysfunction, and cell necrosis and apoptosis. Moreover, increasing evidence suggests that purinergic signaling also mediates the prevention and treatment of MIRI, such as ischemic conditioning, pharmacological intervention, and some other therapies. In conclusion, this review exhibited that purinergic signaling mediates the complex processes of MIRI which shows its promising application and prospecting in the future.

Altered intestinal microbiome and metabolome correspond to the clinical outcome of sepsis.

BACKGROUND: The gut microbiome plays a pivotal role in the progression of sepsis. However, the specific mechanism of gut microbiota and its metabolites involved in the process of sepsis remains elusive, which limits its translational application. METHOD: In this study, we used a combination of the microbiome and untargeted metabolomics to analyze stool samples from patients with sepsis enrolled at admission, then microbiota, metabolites, and potential signaling pathways that might play important roles in disease outcome were screened out. Finally, the above results were validated by the microbiome and transcriptomics analysis in an animal model of sepsis. RESULTS: Patients with sepsis showed destruction of symbiotic flora and elevated abundance of Enterococcus, which were validated in animal experiments. Additionally, patients with a high burden of Bacteroides, especially B. vulgatus, had higher Acute Physiology and Chronic Health Evaluation II scores and longer stays in the intensive care unit. The intestinal transcriptome in CLP rats illustrated that Enterococcus and Bacteroides had divergent profiles of correlation with differentially expressed genes, indicating distinctly different roles for these bacteria in sepsis. Furthermore, patients with sepsis exhibited disturbances in gut amino acid metabolism compared with healthy controls; namely, tryptophan metabolism was tightly related to an altered microbiota and the severity of sepsis. CONCLUSION: Alterations in microbial and metabolic features in the gut corresponded with the progression of sepsis. Our findings may help to predict the clinical outcome of patients in the early stage of sepsis and provide a translational basis for exploring new therapies.

Research on interpersonal violence in schizophrenia: based on different victim types.

BACKGROUND: Schizophrenia is one of the most common severe mental disorders associated with an increased risk of violence. The present study compares the demographical, clinical, and criminological characteristics of the patients with schizophrenia who committed different types of violence to relatives, acquaintances, or strangers. METHOD: Archives of the violent offenders with schizophrenia referred to forensic psychiatric assessments from January 2015 to December 2019 in the West China Forensic Medicine Assessment Center in China were analyzed. The demographic information, mental illness history, and criminological characteristics of the offenders were collected. The clinical symptoms, previous violent behaviors, and social deficits were also evaluated. One-way ANOVA, Kruskal-Wallis test, Chi-square test, and logistic regression analysis were enrolled to do the statistical analysis. RESULTS: The study enrolled 332 cases: 165 cases (49.7%) in the acquaintance victim group (AV), 96 cases (28.9%) in the relative victim group (RV), and 71 cases (21.4%) in the stranger victim group (SV). The multinomial logistic regression analysis revealed that older patients were less likely to attack relatives (OR = 0.966, 95% CI = 0.944-0.990; p = 0.005), and strangers, (OR = 0.971, 95% CI = 0.944-0.998; p = 0.034). Patients who lived with others were more inclined to attack relatives (OR = 15.057, 95% CI = 3.508-64.628; p < 0.001). Additionally, employed patients were more likely to attack strangers (OR = 2.034, 95% CI = 1.036-3.994; p = 0.039). The regression equation did not include psychiatric symptoms. For RV and AV victims, the risk of death was higher compared to that of SV victims (OR = 13.778, p < 0.001; OR = 2.663, p = 0.014). CONCLUSION: In the interpersonal violence cases committed by schizophrenia patients, the victim type correlates with demographic characteristics of offenders such as living situation, age, and employment status, but not with the psychiatric symptoms. The majority of victims were acquaintances and relatives, and the relative victims having more severe injuries. In order to decrease interpersonal violence, especially violent crimes, more people, especially family members and neighbors, should be educated about symptoms of schizophrenia, the ways to communicate with the patients, and the methods for crisis management.

Anophthalmia Caused by Familial Homozygous Mutation of VSX2: a Case Report.

BACKGROUND: Although rare, several mutations in the gene VSX2 (visual system homeobox 2, formerly CHX10) have been associated with congenital autosomal recessive anophthalmia (absence of one or both eyes). This report describes a proband, who at presentation was gravida 2, para 0, and 30 weeks pregnant. METHODS: A 30-year-old woman with congenital bilateral anophthalmia was 30 weeks pregnant at the time of presentation. Her parents were fourth-generation collateral blood relatives, and the familial congenital disease history suggested a possible genetic cause for her anophthalmia. Next generation sequencing and Sanger sequencing of blood samples of the patient, her parents, and her husband were conducted. The fetus was examined via ultrasound. RESULTS: The woman patient had a homozygous variation of the VSX2 gene (NM_182894.2) c.634delC (p.R211 Gfs*90). Both of her parents carried a heterozygous variation of this locus. The husband showed no pathogenic variation in VSX2. The fetal ultrasound revealed bilateral eyeball lenses. A healthy girl was delivered at 41 weeks gestation, with bilateral eyeballs visible. CONCLUSIONS: Homogenous mutation of VSX2 c.634delC (p.R211Gfs*90) has not been reported previously. The patient's congenital bilateral anophthalmia was due to this homogenous mutation, the result of familial inbreeding. Avoiding near-relative marriage is an important means of preventing such diseases.

Intrauterine adhesions combined with Robert's uterus: a case report and literature review.

PURPOSE: To summarize the clinical characteristics and surgical option of Robert's uterus. METHODS: We reported a rare case of Robert's uterus with severe uterine adhesion with successive laparoscopic and hysteroscopic surgery. To our knowledge, such a case has not been reported previously. We also performed a systematic literature review from the PubMed, Embase, and Cochrane databases. RESULTS: Our patient with Robert's uterus with severe uterine adhesions was successfully treated with hysteroscopic septal resection and hysteroscopic adhesiolysis, and the intractable dysmenorrhea disappeared after the hysteroscopic septal resection. In our study, we analyzed the selected 22 reported cases, 10/22 cases (45.5%) were diagnosed before age 20; 20/22 cases (90.91%) experienced dysmenorrhea, 19/22 cases (86.36%) were with hematometra. 5/22 cases (22.73%) underwent re-operation or a third surgery before diagnosis and management. CONCLUSION: Robert's uterus, a rare congenital abnormality of Mullerian duct development, consists of an oblique septum and non-communicating asymmetrical uterine hemi-cavity. The main symptoms are the presence of hematometra and severe dysmenorrhea. Septal resection is the main surgical procedure; however, the rarity and difficulty obtaining a pre-operative diagnosis lead to a high rate of misdiagnosis and second surgery.

Electroacupuncture interventions alleviates myocardial ischemia reperfusion injury through regulating gut microbiota in rats.

Electroacupuncture (EA) intervention has a remarkable cardioprotection against myocardial ischemia reperfusion injury (MIRI). Recently, it has been suggested that the gut microbiota plays an important role in regulating the progression and prognosis of MIRI. The purpose of this study was to illustrate the relationship between gut microbiota and cardioprotection of EA on MIRI. We conducted a MIRI model by ligating the left anterior descending coronary artery for 30 min followed by reperfusion in male Sprague Dawley rats, which then received 7 days of EA intervention. Echocardiography was employed to evaluate left ventricular function. Fecal samples were collected for microbial analysis by 16S rDNA high-throughput sequencing. Blood samples and myocardium were collected for inflammatory cytokine detection by enzyme linked immunosorbent assay (ELISA) and Western blot. Hematoxylin & eosin (HE) staining and immunofluorescence of ileum tissue were performed for intestinal damage evaluation. After 7 days of EA intervention, the left ventricular function was improved with significantly increased ejection fraction and fractional shortening. Furthermore, we found that EA intervention reversed the changed gut microbiota induced by MIRI, including Clostridiales, RF39, S24-7, Desulfovibrio, and Allobaculum, improved the impaired gut barrier, reduced the production and circulation of lipopolysaccharide (LPS), inhibited the level of interleukin 6 (IL-6) and interleukin 12 (IL-12) in periphery and decreased the expression of Toll like receptor 4 (TLR4) and IL-6 in myocardium. EA intervention could improve the impaired gut mucosal barrier and reduce the production and circulation of LPS after MIRI through regulating gut microbiota, thus inhibiting the circulation and myocardium inflammation and finally exerted the cardioprotective effect.

Activation of Src mediates acquired cisplatin resistance in human lung carcinoma cells.

Cisplatin (CDDP) is the most effective chemotherapeutic drug against lung carcinoma. However, the emergence of resistant clones has severely limited its clinical application. We found that the cisplatin-resistant lung carcinoma cell line A549/CDDP had increased levels of the phosphorylated gap junction protein Cx43 and SRC tyrosine kinase, and low levels of total Cx43 protein and reduced gap junction formation. The SRC kinase inhibitor PP2 increased the expression of total Cx43 protein and enhanced cisplatin sensitivity, indicating that activated SRC kinase induces chemoresistance by decrease total Cx43 level. Furthermore, Cx43 gene silencing in the drug-resistant cell lines abrogated the sensitizing effect of PP2. Taken together, targeting SRC kinase by PP2 reverses cisplatin resistance by upregulating Cx43 protein levels, indicating a novel pathway of cisplatin resistance that may be amenable to therapeutic intervention.

Pharmacokinetic/pharmacodynamic analysis of meropenem, by Monte Carlo simulation, in both plasma and cerebrospinal fluid in patients with cerebral hemorrhage after external ventricular drainage
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OBJECTIVE: Patients with cerebral hemorrhage are often prone to intracranial infection, and meropenem is recommended for treatment. But whether the widely used dosing regimen (1 g, 2-hour infusion, every 12 hours) is suitable for antibiotic therapy is still unclear. The purpose of this study was to perform pharmacokinetic/pharmacodynamic (PK/PD) analyses of meropenem in both plasma and cerebrospinal fluid (CSF) in these patients. MATERIALS AND METHODS: Ten patients were enrolled in the present study. The blood samples and CSF samples were taken at predetermined time points and determined by our previously developed HPLC method. Pharmacokinetic parameters were then calculated, and the probability of target attainment (PTA) was calculated by the time that drug concentrations were above the minimum inhibitory concentration (%T>MIC). RESULTS: The peak meropenem concentration (Cmax) of 17.79 ± 3.38 µg/mL in plasma was reached at 2 hours, and the area under the curve (AUC) was 46.95 ± 4.37 h×µg/mL. The Cmax of 6.51 ± 1.11 µg/mL in CSF was reached at 3.50 ± 0.53 hours, and the AUC was 24.53 ± 4.28 h×µg/mL. The average penetration rate of meropenem in these patients was 52.25%. In the case where the MIC value was ≤ 1 µg/mL and using 40%T>MIC as a PK/PD index, the PTA of meropenem in both plasma and CSF were able to provide good coverage with MIC ≤ 1 µg/mL. CONCLUSION: In conclusion, this is the first study on the PK/PD analysis of meropenem in both plasma and CSF in patients with cerebral hemorrhage. The results will assist in selecting appropriate dosing regimens of meropenem in these patients.

Injectable sustained release PLA microparticles prepared by solvent evaporation-media milling technology.

The objective of this study was to develop agomelatine (AGM) intramuscular sustained release PLA microparticles by using solvent evaporation combined with wet milling technology. The final preparation had a regular and homogeneous particle size of approximately 35 µm, as measured by laser diffraction particle size analysis and scanning electron microscopy (SEM). The drug was confirmed to be within the carrier in an amorphous state through differential scanning calorimetry (DSC) and power X-ray diffraction (PXRD) experiments. Additionally, Fourier transform infrared spectroscopy (FT-IR) analysis was applied to confirm that there was hydrogen bonding between the drug and polymer at the molecular level. In vitro release experiments indicated that the drug could achieve long-term sustained release over the period of one month, with only a 3.07% burst release, due to the involvement of the polymer and removal of drug adsorbed on the surface during the wet grinding process. The dominant release mechanism was considered to be diffusion of the drugs in the initial period. Following this, with the hydrolysis of PLA to form a colloidal viscous layer, drug release is due to the combined effect of diffusion and erosion of the polymer matrix. Additionally, drug release behavior is closely related to the degradation mechanism of the polymer carrier. The results suggest that AGM could be developed as a potential delivery system for long-acting intramuscular administration with extensive application prospects.

Pharmacokinetic/Pharmacodynamic Analysis of Meropenem for the Treatment of Nosocomial Pneumonia in Intracerebral Hemorrhage Patients by Monte Carlo Simulation.

BACKGROUND: Nosocomial pneumonia (NP) is a frequent complication among patients with intracerebral hemorrhage (ICH). However, there are currently no pharmacokinetic (PK) and pharmacodynamic (PD) data to guide meropenem dosing in these patients. OBJECTIVE: To investigate the PK/PD properties of meropenem in these patients and whether the usual dosing regimens of meropenem (2-hour infusion, 1 g, every 8 hours) was suitable. METHODS: A total of 11 patients with a diagnosis of ICH complicated with NP were selected in the emergency internal medicine and treated with a 1-g/2-hours extended infusion model. The plasma concentrations of meropenem were determined by high-performance liquid chromatography. PK parameters were estimated by plasma concentration versus time profile using WinNonlin software. The probability of target attainments (PTAs) of meropenem at different minimum inhibitory concentrations (MICs) based on percentage time that concentrations were above the minimum inhibitory concentration (%T>MIC) value were performed by Monte Carlo simulation. RESULTS: The volume of distribution and total body clearance of meropenem were 55.55 L/kg and 22.89 L/h, respectively. Using 40%T>MIC, PTA was >90% at MICs ≤4 µg/mL. Using 80% or 100%T>MIC, PTA was >90% only at MICs ≤1 µg/mL. CONCLUSIONS: The PK/PD profile of dosing regimens tested will assist in selecting the appropriate meropenem regimens for these patients. At a target of 40%T>MIC, the usual dosing regimens can provide good coverage for pathogens with MICs of ≤4 µg/mL. However, when a higher target (80% or 100%) is desired for difficult-to-treat infections, larger doses, prolonged infusions, shorter intervals, and/or combination therapy may be required.

Elevated Preoperative Carcinoembryonic Antigen and Vascular Endothelial Growth Factor Predict Shorter Survival in Patients with Sigmoid Colon Carcinoma.

BACKGROUND: We investigated the prognostic significance of carcinoembryonic antigen (CEA) and vascular endothelial growth factor (VEGF) and their associations with clinicopathological features in patients with sigmoid colon carcinoma (SCC). METHODS: We retrospectively reviewed patients with SCC treated with curative surgery from January 2001 to June 2010 at our hospital. Patients' general, clinical, histopathologic, and serum biomarkers were analyzed. We measured VEGF in 176 sets of SCC tissues and adjacent noncancerous tissue samples with immunohistochemical and Elivison staining. CEA was measured with a tumor biomarker chip. Their correlations with clinicopathologic factors were analyzed by chi-square test. Univariate and multivariate analyses were used to identify factors associated with overall survival (OS). RESULTS: Of the 176 patients, 69.3% were alive and 30.7% had died. In univariate analysis, serum CEA level (p = 0.002, OR = 2.394, 1.392 - 4.116), tumor VEGF (p = 0.04, OR = 1.968, 1.032 - 3.752), lymph node metastasis (N, p = 0.000, OR = 3.712, 2.064 - 6.675), T stage (T, p = 0.016, OR = 5.706, 1.382 - 23.552) and differentiation (p = 0.000) were the prognostic factors for OS. In stratified analysis, combined elevated serum CEA and tumor VEGF levels were associated with poorer prognosis. CONCLUSIONS: Elevated preoperative serum CEA and tumor VEGF were predictors of poor prognosis for patients with SCC.

Identification and expression analysis of microRNAs during ovule development in rice (Oryza sativa) by deep sequencing.

KEY MESSAGE: MicroRNA (miRNA) expression profiles during rice ovule development revealed the possible miRNA-mediated regulation between ovule sporophytic tissue and female gametophyte and the involvement of miRNAs in programmed cell death. MiRNAs are 20-24-nucleotide small RNAs that play key roles in the regulation of many growth and developmental processes in plants. Rice ovule development comprises a series of biological events, which are regulated by complex molecular mechanisms. To gain insight into miRNA-mediated regulation of rice ovule development, Illumina sequencing was used to examine the expression of miRNAs from the megaspore mother cell meiosis stage to the fertilized ovule stage. Based on the sequencing data, 486 known and 204 novel miRNAs were identified during rice ovule development. Moreover, 56, 65 and 11 differentially expressed miRNAs between adjacent developmental stages were identified. By analyzing transcriptome and degradome data, we identified 41, 65 and 12 coherent target genes for the differentially expressed miRNAs in ovule development. We found that changes in the expression of plant hormone-related miRNAs may play important roles in embryo sac development, providing evidence for cross-talk communication between sporophytic tissue and the female gametophyte. Additionally, we revealed that miRNAs may be involved in programmed cell death after fertilization. Finally, we constructed miRNA-mediated regulatory networks that are active during rice ovule development.

Impact of VKORC1 gene polymorphisms on warfarin maintenance dosage: A novel systematic review and meta-analysis of 53 studies.

OBJECTIVE: To analyze the correlation between VKORC1 gene polymorphisms and warfarin maintenance dosage, as well as the correlation of dosage of warfarin with age and ethnicity. METHODS: We retrieved related studies published between January 2000 and March 2016 from PubMed, Embase, the Cochrane Library, Web of Science, VIP, CNKI, and Wan Fang data. Two reviewers independently screened literature according to the inclusion and exclusion criteria, extracted data and crosscheck data. Then, RevMan5.3 software was used to perform a meta-analysis. RESULTS: 53 studies were included in the meta-analysis. The most prevalent genotypes were -1639 AA, 1173 TT, and 3730 GG in both Asian and Caucasians, but the distribution frequencies of all three were higher in Asians than in Caucasians. The meta-analysis showed that compared with homozygous VKORC1-1639 AA carriers, carriers of type GA, GG, and G (GA + GG) required 45% (95% confidence interval (CI) 42 - 49), 77% (95% CI 70 - 84), and 51% (95% CI 47 - 55) higher warfarin doses, respectively. Carriers of type CC, TC, and C (CC + TC) required 83% (95% CI 73 - 92), 26% (95% CI 23 - 29), and 53% (95% CI 44 - 62) higher warfarin doses, respectively, compared to homozygous VKORC1 1173 TT carriers. Carriers of type AA, GA, and A (AA + GA) required 40% (95% CI 29 - 51), 25% (95% CI 17 - 33), and 33% (95% CI 21 - 45) higher warfarin doses, respectively, compared to carriers of the homozygous VKORC1 3730 GG polymorphism (all p < 0.05). Subgroup analysis showed that Asian patients aged ≤ 60 years carrying 1173 CC, TC, and C genotypes required 28%, 39%, and 22% higher warfarin doses, respectively, compared with patients aged > 60 years. Caucasian patients aged > 60 years carrying -1639 GA, GG and G genotypes needed 24%, 39%, and 37% lower warfarin doses, respectively, compared with patients aged ≤ 60 years. These differences were statistically significant (p < 0.05). CONCLUSIONS: Our study showed that the relationship between VKORC1 gene polymorphisms and warfarin maintenance dose differs between individuals, and that individuals with different ages and ethnicities require different doses of warfarin. Caucasians carriers of genotype -1639 GG, G and 1173 CC, TC, C required a higher mean daily warfarin doses compared with Asian patients. Therefore, in order to achieve optimal treatment and lowest risk, VKORC1 gene polymorphism detection is suggested.
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Genome-wide transcriptome analysis of female-sterile rice ovule shed light on its abortive mechanism.

MAIN CONCLUSION: The comprehensive transcriptome analysis of rice female-sterile line and wild-type line ovule provides an important clue for exploring the regulatory network of the formation of rice fertile female gametophyte. Ovules are the female reproductive tissues of rice (Oryza sativa L.) and play a major role in sexual reproduction. To investigate the potential mechanism of rice female gametophyte fertility, we used RNA sequencing, combined with genetic subtraction, to compare the transcriptome of the ovules of a high-frequency female-sterile line (fsv1) and a rice wild-type line (Gui 99) during ovule development. Ovules were harvested at three developmental stages: ovule containing megaspore mother cell in meiosis process (stage 1), ovule containing functional megaspore in mitosis process (stage 2), and ovule containing mature female gametophyte (stage 3). Six cDNA libraries generated a total of 42.2 million high-quality clean reads that aligned with 30,204 genes. The comparison between the fsv1 and Gui 99 ovules identified a large number of differentially expressed genes (DEGs), i.e., 45, 495, and 932 DEGs at the three ovule developmental stages, respectively. From the comparison of the two rice lines, Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and MapMan analyses indicated that a large number of DEGs associated with starch and sucrose metabolism, plant hormone signal transduction, protein modification and degradation, oxidative phosphorylation, and receptor kinase. These DEGs might play roles in ovule development and fertile female gametophyte formation. Many transcription factor genes and epigenetic-related genes also exhibit different expression patterns and significantly different expression levels in two rice lines during ovule development, which might provide important information regarding the abortive mechanism of the female gametophyte in rice.

Directed chromosomal integration and expression of porcine rotavirus outer capsid protein VP4 in Lactobacillus casei ATCC393.

Using two-step plasmid integration in the presence of 5-fluorouracil (5-FU), we developed a stable and markerless Lactobacillus casei strain for vaccine antigen expression. The upp of L. casei, which encodes uracil phosphoribosyltransferase (UPRTase), was used as a counterselection marker. We employed the Δupp isogenic mutant, which is resistant to 5-FU, as host and a temperature-sensitive suicide plasmid bearing upp expression cassette as counterselectable integration vector. Extrachromosomal expression of UPRTase complemented the mutated chromosomal upp allele and restored sensitivity to 5-FU. The resultant genotype can either be wild type or recombinant. The efficacy of the system was demonstrated by insertion and expression of porcine rotavirus (PRV) VP4. To improve VP4 expression, we analyzed L. casei transcriptional profiles and selected the constitutive highly expressed enolase gene (eno). The VP4 inserted after the eno termination codon were screened in the presence of 5-FU. Using genomic PCR amplification, we confirmed that VP4 was successfully integrated and stably inherited for at least 50 generations. Western blot demonstrated that VP4 was steadily expressed in medium with different carbohydrates. RT-qPCR and ELISA analysis showed that VP4 expression from the chromosomal location was similar to that achieved by a plasmid expression system. Applying the recombinant strain to immunize BALB/c mice via oral administration revealed that the VP4-expressing L. casei could induce both specific local and systemic humoral immune responses in mice. Overall, the improved gene replacement system represents an efficient method for chromosome recombination in L. casei and provides a safe tool for vaccine production.

A systematic review and meta-analysis of acupuncture for improving learning and memory ability in animals.

BACKGROUND: Memory loss is the most prominent symptoms of brain aging, but there is currently no evidence-based treatment strategy. Acupuncture has been widely used in China and the effectiveness for improving learning and memory has been mentioned in previous studies. We conducted this systematic review and meta-analysis to evaluate the effectiveness of acupuncture for improving learning and memory in animal experiments. METHODS: We searched Pubmed, Embase, Ovid Medline(R), the China National Knowledge Infrastructure (CNKI), Chinese Science and Technology Periodical Database (VIP) and Wanfang data Information Site to collect studies published up to December 2015. Study quality for each included article was evaluated according to the CAMARADES 10-item checklist. Outcome measure is Morris water maze. A meta-analysis was conducted according to the Cochrane systematic review method by using RevMan 5.3 software. RESULTS: Forty-two studies involving 944 animals were included. The quality score of the studies ranged from 2 to 8, with a mean of 5.3. Meta-analysis results showed that 24 studies reported significant effect of acupuncture for decreasing escape latency (-3.00, 95 % CI: -3.78 ~ -2.23, P < 0.00001), 14 studies reported significant effect of acupuncture for increasing frequency of cross platform (2.57, 95 % CI: 1.92 ~ 3.22, P < 0.00001), and 7 studies reported significant effect of acupuncture for increasing time in target quadrant (2.00, 95 % CI: 1.10 ~ 2.91, P < 0.00001) compared with the control group. CONCLUSIONS: These findings show acupuncture has a potential role in improving learning and memory ability in animal models, suggesting it as a candidate therapy for memory loss of aged brain.

[The dynamic changes of tuberculosis related cytokines during disease progression].

OBJECTIVE: To study the dynamic changes of tuberculosis related cytokines among patients during the different courses of treatment, and to analyze their influences on the development and prognoses of tuberculosis. METHODS: All patients with active tuberculosis were enrolled from Guangzhou, Shenzhen and Foshan TB control institutes. There were a total of 68 cases, 36 males and 32 females, aged 19 to 50 years [ average (30±9) years]. All the TB patients received standard chemotherapy regimen of anti-tuberculosis, and were divided into 2 groups: one completed treatment group (cured or clinically cured 38 cases) and 1 uncompleted treatment group (treatment failure or need to extend treatment, 30 cases). Peripheral blood serum at 0, 2, 6 month during the treatment from 68 tuberculosis patients were collected, and the concentration of IFN-γ,IL-4,IL-17,TGF-ß,TNF-α and IL-10 were detected by ELISA tests. RESULTS: The concentration of IFN-γ, TGF-ß and IL-4 in all enrolled patients showed significant decrease (from 23.2 ng/L to 22.3 ng/L, from 169.1 ng/L to 123.2 ng/L; 65.0 ng/L to 31.9 ng/L) (t=2.67, 2.35 and 3.41, P<0.05) along with the extension of treatment. IL-10 increased significantly (12.9 ng/L) in the uncompleted treatment group but declined significantly (5.38 ng/L) (P<0.05) in the completed treatment group at the end of 6 month. Meanwhile, IL-4 decreased significantly (P<0.05) in the completed treatment group but no significant changes were observed in the uncompleted treatment group. Th1/Th2 (IFN-γ/IL-4) raised gradually in the completed treatment group (0 month <2 month <6 month, t=6.32, 6.03 and 5.85, P<0.05), while it was only at 6 month in the uncompleted treatment group (0 month <6 month, t=3.7, P<0.05). And the ratio of Th1/Th2 in the completed treatment group was significantly higher than that in the uncompleted group treatment (P<0.05). CONCLUSION: It suggests that the changes of Th1 cytokines (IFN-γ, TGF-ß) and the Th1/Th2 balance play an important role in the pathogenesis, development and prognosis of TB. The suppression of IFN-γ, TGF-ß or Th1/Th2 balance may be an important factor influencing the prognosis of TB.