[Clinical characteristics and genetic analysis of a patient with Acephalic spermatozoa syndrome due to variant of PMFBP1 gene].

OBJECTIVE: To analyze the clinical characteristics and genetic basis of a male patient with primary infertility caused by Acephalic spermatozoa syndrome. METHODS: A patient who had presented at the Henan Provincial People's Hospital on October 1, 2022 was selected as the study subject. Clinical data and results of laboratory exams and sperm electron microscopy were collected. The patient was subjected to whole exome sequencing (WES), and candidate variants were verified by Sanger sequencing and pathogenicity analysis. RESULTS: WES revealed that the patient has harbored compound heterozygous variants of the PMFBP1 gene, namely c.853del (p.Ala285Leufs*24) and c.1276A>T (p.Lys426X), which were both unreported previously. Sanger sequencing suggested that the c.853del (p.Ala285Leufs*24) variant has derived from his deceased mother, whilst the c.1276A>T (p.Lys426X) variant has derived from his father. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), both variants were classified as pathogenic (PVS1+PM2_Supporting+PP4). CONCLUSION: The compound heterozygous variants of the PMFBP1 gene probably underlay the Acephalic spermatozoa syndrome in this patient. The discovery of the novel variants has also enriched the mutational spectrum of Acephalic spermatozoa syndrome.

Comparison between effects of mini-implant anchorage and face-bow anchorage in orthodontics for children.

To study the values of mini-implant anchorage in orthodontics for children in the mixed dentition stage, 78 children in the mixed dentition stage who had accepted orthodontic treatment in our hospital from January 2020 to January 2021 were enrolled into this study. All children were treated with straight-wire appliance. According to their anchorages, children were divided into observation group and control group based on the random number table. Children in the control group used face-bow to control the anchorages and children in the observation group used mini-implants to control the anchorages. After treatment, the upper central incisor convex distance difference, inclination angle of the upper central incisor, displacement of the molar, gingival health, masticatory function, treatment effect and adverse reaction rate of children in two groups were compared. One year after treatment, compared with children in the control group, children in the observation group had smaller the upper central incisor convex distance difference, inclination angle of the upper central incisor, displacement of the molar, small scores of plaque index (PLI), bleeding index (BI) and gingival index (GI), stronger biting force and higher masticatory efficiency, lower adverse reaction rate during treatment, better treatment effect, higher satisfaction of orthodontic treatment. And differences of all the above indexes were statistically significant (p < 0.05). Mini-implant anchorages have good stability and directive force, and have certain values in orthodontics for children in the mixed dentition stage.

A novel mutation in PCD-associated gene DNAAF3 causes male infertility due to asthenozoospermia.

Primary ciliary dyskinesia (PCD) is a rare autosomal-recessive disease manifested with recurrent infections of respiratory tract and infertility. DNAAF3 is identified as a novel gene associated with PCD and different mutations in DNAAF3 results in different clinical features of PCD patients, such as situs inversus, sinusitis and bronchiectasis. However, the sperm phenotypic characteristics of PCD males are generally poorly investigated. Our reproductive medicine centre received a case of PCD patient with infertility, who presented with sinusitis, recurrent infections of the lower airway and severe asthenozoospermia; However, no situs inversus was found in the patient. A novel homozygous mutation in DNAAF3(c.551T>A; p.V184E) was identified in the PCD patient by whole-exome sequencing. Subsequent Sanger sequencing further confirmed that the DNAAF3 had a homozygous missense variant in the fifth exon. Transmission electron microscopy and immunostaining analysis of the sperms from the patient showed a complete absence of outer dynein arms and partial absence of inner dynein arms, which resulted in the reduction in sperm motility. However, this infertility was overcome by intracytoplasmic sperm injections, as his wife achieved successful pregnancy. These findings showed that the PCD-associated pathogenic mutation within DNAAF3 also causes severe asthenozoospermia and male infertility ultimately due to sperm flagella axoneme defect in humans. Our study not only contributes to understand the sperm phenotypic characteristics of patients with DNAAF3 mutations but also expands the spectrum of DNAAF3 mutations and may contribute to the genetic diagnosis and therapy for infertile patient with PCD.

[Analysis of DNAH1 gene variant in two infertile males with multiple morphological abnormalities of sperm flagella].

OBJECTIVE: To explore the clinical feature and gene variant for two cases of primary male infertility caused by severe asthenospermia and to analyze the etiology of the disease. METHODS: Genomic DNA of peripheral blood samples of patients and their parents was extracted and gene variant analysis of the patients was conducted by using whole exome sequencing. Suspected pathogenic variant was verified by Sanger sequencing and pathogenic analysis. RESULTS: Whole exome sequencing showed that the DNAH1 gene of patient 1 had two heterozygous variants of c.2016T>G(p.Y672X) and c.6017T>G (p.V2006G). The DNAH1 gene of patient 2 had a homozygous variant of c.2610G>A(p.W870X), which were inherited from his father and mother, respectively. According to American College of Medical Genetics and Genomics standards and guidelines, the c.2016T>G (p.Y672X) and c.2610G>A (p.W870X) varaints of DNAH1 gene were predicted to be pathogenic (PVS1+PM2+PM3+PP3). CONCLUSION: The two patients of multiple morphological abnormalities of the sperm flagella may be caused by DNAH1 gene variant, which has resulted in primary male infertility.

[Sperm-hyaluronan binding assay in in vitro fertilization and embryo transfer].

OBJECTIVE: To explore the application value of sperm-hyaluronan binding (SHB) assay in in vitro fertilization and embryo transfer (IVF-ET). METHODS: This retrospective study included 163 cases of IVF-ET performed in our hospital from January to August 2019 due to female fallopian tube abnormality-induced infertility. The men were found with normal seminal parameters at semen analysis. According to the SHB rate, we divided the patients into a normal group (n = 126) and an abnormal group (n = 37) and analyzed the general conditions and the rates of normal fertilization, cleavage, available embryos, embryo implantation and clinical pregnancy. RESULTS: Statistically significant differences were observed between the normal and abnormal groups in the rates of SHB, normal fertilization and available embryos (P < 0.01 or P < 0.05), but not in the general conditions or the rates of cleavage, embryo implantation and clinical pregnancy. CONCLUSIONS: The SHB rate is not correlated with sperm concentration or the percentages of progressively motile sperm and morphologically normal sperm. Sperm-hyaluronan binding assay helps predict the outcomes of IVF-ET and embryo quality. An SHB rate of <58.5% indicates low rates of normal fertilization and available embryos, but has no significant correlation with the rates of embryo implantation and clinical pregnancy.

[Effects of Yishen Shengjing Capsules on semen quality and gonadal hormone levels in rats with dibutyl phthalate-induced reproductive function injury].

OBJECTIVE: To study the possible pathogenesis of infertility caused by dibutyl phthalate (DBP) and investigate the effects of Yishen Shengjing Capsules (YSC, kidney-tonifying and essence-producing capsules) on DBP-induced reproductive function injury and its possible action mechanisms in male Wistar rats. METHODS: Models of DBP-induced reproductive function injury were made in 80 male Wistar rats and another 20 were used as blank controls. After modeling, the model rats were randomly divided into a model control, a high-dose YSC, a medium-dose YSC, and a low-dose YSC group. Four weeks after intervention, all the animals were sacrified for observation of the histomorphologic changes in the testis under the light microscope, measurement of sperm concentration, motility and abnormality, and determination of the levels of serum testosterone (T), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) by radioimmunoassay. RESULTS: Compared with the blank controls, the model rats showed obvious pathological changes in testicular histomorphology, significantly decreased sperm concentration and motility, increased sperm abnormality, reduced contents of serum T and LH, and elevated the level of serum FSH (P<0.01). After 4 weeks of medication, the animals of the high-, medium-, and low-dose YSC groups, in comparison with the model controls, exhibited different degrees of recovery from testicular histomorphological damage, remarkably increased sperm concentration and motility, decreased sperm abnormality, elevated levels of serum T and LH, and reduced content of serum FSH (P<0.01). There were statistically significant differences in all the parameters above between the high-dose YSC and medium- and low-dose YSC groups (P<0.01). CONCLUSIONS: DBP reduces sperm motility and concentration, increases sperm abnormality, causes damage to the morphological structure of the rat testis, decreases the contents of serum T and LH, and elevates the level of the serum FSH. Yishen Shengjing Capsules can improve DBP-induced productive function injury, increase sperm motility and concentration, decrease sperm abnormality, elevate the level of serum T and LH, reduce the content of serum FSH, improve the morphological structure of the testis, and thus promote the reproductive function of the male rat.

[Inhibitory effect of Qiangjing Tablets on the Fas/FasL pathway of cell apoptosis in male SD rats with infertility].

OBJECTIVE: To observe the effects of Qiangjing Tablets (QJT) on the semen quality and Fas/FasL signaling pathway in male SD rats with infertility. METHODS: Models of infertility were made in 50 male SD rats by intragastric administration of Tripterygium (GTW) for 3 weeks, and another 20 rats were taken as blank controls. Then 40 successfully established rat models were randomly divided into four groups, model control, low-dose QJT, medium-dose QJT, and high-dose QJT, the latter three groups treated intragastrically with QJT at 58 mg, 105 mg, and 233 mg per kg of the body weight per day, respectively. After 4 weeks of medication, the rats were killed for examination of semen quality and determination of the expression of the apoptosis factor FasL in the testis tissue. RESULTS: Compared with the blank controls, the model rats showed significant decreases in sperm concentration ([71.99 ± 9.72] vs [10.94 ± 3.58] x 106/ml, P < 0.01), motility ([48.95 ± 4.10] vs [9.31 ± 5.79]%, P < 0.01), and viability ( [82.06 ± 6.16] vs [24.03 ± 6.93]%, P < 0.01). In comparison with the model controls, the rats in the QJT groups exhibited remarkably increased sperm concentration, motility, and viability, more significantly in the high-dose group ([59.66 ± 4.53] x 106/ml, [35.45 ± 5.21] %, and [61.97 ± 9.75]%) and medium-dose group ([40.89 ± 4.90] x 106/ml, [24.41 ± 4.79]%, and [60.06 ± 10.62]%) (P < 0.05 or P < 0.01). The expression of FasL was markedly reduced in the low-, medium-, and high-dose QJT groups (0.5215 ± 0.0189, 0.5371 ± 0.0364, and 0.4556 ± 0.0215) as compared with that of the model controls (0.5989 ± 0.0448 ) (P < 0.05 or P < 0.01). CONCLUSION: By upregulating the Fas/FasL signaling pathway, Tripterygium glycosides may induce the apoptosis of spermatogenic cells and reduce sperm concentration, motility and viability, resulting in infertility. The Chinese medicine Qiangjing Tablets can improve the reproductive function of male rats by decreasing the expression of the apoptosis factor FasL in the testis.

[Expression and clinical significance of transcription factors in prostate cancer].

OBJECTIVE: To explore the roles of various transcription factors in the occurrence and development of prostate cancer. METHODS: A total of 139 specimens with prostate cancer (PCa) and 83 specimens with benign prostatic hyperplasia (BPH) from hospitalized patients at our hospital from 2008 to 2011 were enrolled. The mRNA expressions of c-Myc, Klf4, Nanog, Oct4A and Sox2 were determined by quantitative real-time polymerase chain reaction (PCR) and the expressions of Klf4 isoforms by conventional PCR. Immunohistochemical method was used for the detection of Klf4 protein via tissue microarray in 404 prostate samples. RESULTS: No significant difference existed in the expressions of Nanog, Oct4A and Sox2 genes between BPH and PCa samples. And the expressions of c-Myc and Klf4 genes were significantly higher in PCa than those in BPH specimens. Immunohistochemical results showed that Klf4 protein could be detected in a large majority of epithelial prostatic cells irrespective of malignant transformation. However, it was predominantly located cytoplasmically in PCa tissues and remained consistent with the expression of a differentially spliced Klf4α isoform. CONCLUSION: Klf4 is highly expressed in both BPH and PCa tissues. But in malignant cells, a specific gene product Klf4α is predominantly detectable in cytoplasm. The positioning of Klf4 protein may have an important relationship with its role in the tumorigenic process.

[Clinical significance of serum telomerase mRNA in patients with prostate cancer].

OBJECTIVE: To explore the clinical significance of detecting serum mRNA of telomerase in the diagnosis of prostate cancer. METHODS: Serum samples were collected from 29 patients with prostate cancer and 29 age-matched patients with cardiovascular or metabolic disease as non-tumor controls. Sera samples from 15 healthy age-matched subjects were used as healthy control. Detection of serum telomerase mRNA was performed with real-time reverse transcriptase quantitative polymerase chain reaction (PCR). RESULTS: Serum telomerase mRNA was detectable in 89.7% (26/29) patients with prostate cancer, but almost undetectable in non-tumor (6.9%, 2/29) and healthy control groups(1/14). CONCLUSION: Using real-time PCR for detecting serum telomerase mRNA may be an auxiliary method for diagnosing and monitoring of prostate cancer.

Nitrogen-Doped Carbon Quantum Dots with Photoactivation Properties for Ultraviolet Ray Detection.

Photoactivation is a phenomenon that could enhance the photoluminescence (PL) and photostability upon UV/vis light exposure, which is usually observed in CdSe/ZnS quantum dots (QDs). However, the photoactivation phenomenon has been scarcely reported in fluorescent carbon quantum dots (CQDs). Herein, the nitrogen-doped carbon quantum dots (N-CQDs) were prepared through a facile solvothermal approach with naphthalenetracarboxylic dianhydride and serine as precursors. Upon simple UV light irradiation for 10 min, the fluorescence quantum yield (QY) of N-CQDs could increase up to 10-fold. Based on this phenomenon, the N-CQDs were explored as an ultraviolet (UV) light sensor to assess the intensity of ultraviolet radiation in sunlight and indirectly evaluate the UV-blocking efficiency of various sunscreen products. Thus, this contribution not only provided an insight into developing a low-cost UV detector but also opened a door for the development of carbon quantum dots with converse-photobleaching properties.

Compound heterozygous mutations in CFTR causing congenital bilateral absence of the vas deferens in a Chinese pedigree.

BACKGROUND: Cystic fibrosis (CF) is an autosomal recessive disorder rarely found in Asian populations. Most males with CF are infertile because of obstructive azoospermia (OA) caused by congenital bilateral absence of the vas deferens (CBAVD). Compound heterozygous mutations of cystic fibrosis transmembrane conductance regulator (CFTR) are among the most common pathogenic factors in CBAVD. However, few genealogical analyses have been performed. METHODS: In this study, whole-exome sequencing and cosegregation analysis were performed in a Chinese pedigree involving two siblings with CBAVD. Moreover, in vitro gene expressions were used to analyze the pathogenicity of a novel CFTR mutation. RESULTS: We identified compound heterozygous mutations of CFTR comprising the known disease-causing variant c.1210-11T>G (also known as IVS9-5 T) and c.2144delA;p.q715fs in two siblings with CBAVD. To verify the effects in vitro, we transfected vectors expressing wild-type and mutated CFTR into 293T cells. The results showed that the CFTR protein containing the frameshift mutation (c.2144delA) was 60 kD smaller. With testicular sperm aspiration/intracytoplasmic sperm injection-embryo transfer (TESA/ICSI-ET), both CBAVD patients fathered healthy offspring. CONCLUSION: Our study revealed that compound heterozygous mutations of CFTR are involved in CBAVD, expanding the known CFTR gene mutation spectrum of CBAVD patients and providing more evidence that compound heterozygous mutations can cause familial CBAVD.

Robust PANI@MXene/GQDs-Based Fiber Fabric Electrodes via Microfluidic Wet-Fusing Spinning Chemistry.

Two-dimensional transition metal titanium carbide (Ti3 C2 Tx ) as a promising candidate material for batteries and supercapacitors has shown excellent electrochemical performance, but it is difficult to meet practical applications because of its poor morphology structure, low mechanical properties, and expensive process. Here, an applied and efficient method based on microfluidic wet-fusing spinning chemistry (MWSC) is proposed to construct hierarchical structure of MXene-based fiber fabrics (MFFs), allowing the availability of MFF electrodes with ultrastrong toughness, high conductivity, and easily machinable properties. First, a dot-sheet structure constructed by graphene quantum dots (GQDs) and MXene nanosheets with multianchor interaction in the microchannel of a microfluidic device enhances the mechanical strength of MXene fibers; next, the interfused fiber network structure of Ti3 C2 Tx /GQDs fabrics assembled by the MWSC process enhances the deformability of the whole fabrics; finally, the core-shell structure of PANI@Ti3 C2 Tx /GQDs architected by in-situ polymerization growth of polyaniline (PANI) nanofibers provides more ion-accessible pathways and sites for kinetic migration and ion accumulation. Through the morphology and microstructure design, this strategy has directive significance to the large-scale preparation of conductive fabric electrodes and provides a viable solution for simultaneously enhancing mechanical strength and electrochemical performance of conductive fabric electrodes.

Correlation between CFTR variants and outcomes of ART in patients with CAVD in Central China.

Biallelic variants in Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) are the main pathogenic factor of congenital absence of the vas deferens (CAVD), including congenital bilateral absence of the vas deferens (CBAVD) and congenital unilateral absence of the vas deferens (CUAVD). However, there are few reports about the correlation between CFTR variant and outcomes of assisted reproductive technology (ART) in CAVD patients of China. In this study, 104 patients with CAVD were recruited in Central China, and provided gene detection by the whole-exome sequencing, among them 69% (72/104) carried at least one variant in CFTR and one carried adhesion G protein-coupled receptor G2 (ADGRG2) variant. A total of 81 CAVD patients were treated with ART, of which 21 and 60 carried none or at least one variant in CFTR, respectively. The fertilization rate, cleavage rate, effective embryo rate, implantation rate, clinical pregnancy rate and live birth rate per fresh embryo transfer were compared between patients with and without CFTR variants. It was found that the ART outcomes had no significant difference whether the patients carried the CFTR variant or not. In addition, all of the offspring were healthy after follow-up. In conclusion, rare CFTR variants may play a major role in patients with CAVD in Central China, which were greatly different from other descent. There was no significant difference in ART outcomes in CAVD patients with or without CFTR variants. The limitations of this study were that there was no statistical analysis of the sperm quality through TESA and conclusions were relatively limited due to the small sample size of the study.

Potential mechanisms of osthole against bladder cancer cells based on network pharmacology, molecular docking, and experimental validation.

BACKGROUND: Osthole was traditionally used in treatment for various diseases. However, few studies had demonstrated that osthole could suppress bladder cancer cells and its mechanism was unclear. Therefore, we performed a research to explore the potential mechanism for osthole against bladder cancer. METHODS: Internet web servers SwissTargetPrediction, PharmMapper, SuperPRED, and TargetNet were used to predict the Osthole targets. GeneCards and the OMIM database were used to indicate bladder cancer targets. The intersection of two target gene fragments was used to obtain the key target genes. Protein-protein interaction (PPI) analysis was performed using the Search Tool for the Retrieval of Interacting Genes (STRING) database. Furthermore, we used gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses to explore the molecular function of target genes. AutoDock software was then used to perform molecular docking of target genes,osthole and co-crystal ligand. Finally, an in vitro experiment was conducted to validate bladder cancer inhibition by osthole. RESULTS: Our analysis identified 369 intersection genes for osthole, the top ten target genes included MAPK1, AKT1, SRC, HRAS, HASP90AA1, PIK3R1, PTPN11, MAPK14, CREBBP, and RXRA. The GO and KEGG pathway enrichment results revealed that the PI3K-AKT pathway was closely correlated with osthole against bladder cancer. The osthole had cytotoxic effect on bladder cancer cells according to the cytotoxic assay. Additionally, osthole blocked the bladder cancer epithelial-mesenchymal transition and promoted bladder cancer cell apoptosis by inhibiting the PI3K-AKT and Janus kinase/signal transducer and activator of transcription (JAK/STAT3) pathways. CONCLUSIONS: We found that osthole had cytotoxic effect on bladder cancer cells and inhibited invasion, migration, and epithelial-mesenchymal transition by inhibiting PI3K-AKT and JAK/STAT3 pathways in in vitro experiment. Above all, osthole might have potential significance in treatment of bladder cancer. SUBJECTS: Bioinformatics, Computational Biology, Molecular Biology.

A Novel Noncanonical Splicing Mutation of ANOS1 Gene in Siblings with Kallmann Syndrome Identified by Whole-Exome Sequencing.

Kallmann syndrome (KS) is a rare genetic disorder that is characterized by idiopathic hypogonadotropic hypogonadism associated with anosmia. Genetic variants in ANOS1 gene are the most common mutations associated with X-linked recessive form of KS. Canonical ± 1 or 2 splice site variants in ANOS1 have been described to be responsible for KS. Here, we identified a novel noncanonical splice site variant (c.1062+4T>C) in ANOS1 gene in two siblings with KS by whole-exome sequencing (WES). Sanger sequencing showed this mutation was inherited from their mother, whose brother was a KS patient as well. Through the functional assay in vitro, we found that this mutation resulted in a 50-bp deletion of exon 7, which caused frameshift mutation leading to a premature termination of translation and a truncated anosmin-1 protein. Our results revealed that this noncanonical splice site variant is involved in KS. Thus, it is suggested that we should pay attention to the noncanonical splice site variants when using molecular genetic diagnostics of KS.

Identification of a six-gene prognostic signature for bladder cancer associated macrophage.

As major components of the tumor microenvironment (TME), tumor-associated macrophages (TAMs) play an exceedingly complicated role in tumor progression and tumorigenesis. However, few studies have reported the specific TAM gene signature in bladder cancer. Herein, this study focused on developing a TAM-related prognostic model in bladder cancer patients based on The Cancer Genome Atlas (TCGA) data. Weighted Gene Co-Expression Network Analysis (WGCNA) was used to identify key genes related to TAM (M2 macrophage). Gene ontology (GO) enrichment and the Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis showed the functional categories of the key genes. Simultaneously, we used the Least Absolute Shrinkage and Selection Operator (LASSO) and univariate and multivariate Cox regressions to establish a TMA-related prognostic model containing six key genes: TBXAS1, GYPC, HPGDS, GAB3, ADORA3, and FOLR2. Subsequently, single-cell sequencing data downloaded from Gene Expression Omnibus (GEO) suggested that the six genes in the prognostic model were expressed in TAM specifically and may be involved in TAM polarization. In summary, our research uncovered six-TAM related genes that may have an effect on risk stratification in bladder cancer patients and could be regarded as potential TAM-related biomarkers.

[Effects of Qiangjing Capsule on the oxidative and antioxidative system in the epididymis of varicocele rats].

OBJECTIVE: To observe the effects of Qiangjing Capsule (QC) on the oxidative amd antioxidative system in the epididymis of varicocele rats in comparison with those of Shaofuzhuyu Capsule (SC) and Wuziyanzong Capsule (WC), and to explore its possible mechanism of enhancing epididymal sperm maturation. METHODS: Ten of 100 adolescent male SD rats were randomized to a sham-operation group, and varicocele models were successfully established in 72 of the other 90 by narrowing of the left renal vein. Then the model rats were equally assigned to 6 groups: model control, high-dose QC (0.216 g/ml), medium-dose QC (0.108 g/ml), low-dose QC (0.054 g/ml), SC (0.146 g/ml), and WC (0.130 g/ml). After 4 weeks of treatment, we determined the activity of glutathione peroxidase (GPx) and the level of malondialdehyde (MDA) in the left epididymis of different groups of rats. RESULTS: Compared with the sham-operation group, the model group showed a significant decrease in GPx activity (P < 0.01) and a marked increase in the MDA level (P < 0.05). And the high-dose QC group exhibited a significantly hither GPx activity and lower MDA level than all the other groups (P < 0.01 and P < 0.05). CONCLUSION: Varicocele can reduce the activity of GPx and elevate the level of MDA in the epididymis of rats, while Qiangjing Capsule can increase the former and decrease the latter, and thereby may improve epididymal microenvironment, enhance epididymal sperm maturation and promote fertility.

Preparation of species-specific monoclonal antibody and development of fluorescence immunoassay based on fluorescence resonance energy transfer of carbon dots for accurate and sensitive detection of Alicyclobacillus acidoterrestris in apple juice.

The growth and metabolism of Alicyclobacillus acidoterrestris can lead to the spoilage of commercial fruit juice. Existing methods have some drawbacks such as complex sample pretreatment, skilled technician requirement, reduced sensitivity and specificity. Herein, a novel fluorescence immunoassay was developed using a monoclonal antibody (mAb) against A. acidoterrestris as the sensing element and carbon dots (CDs) as the signal response unit. The CDs can be quenched via fluorescence resonance energy transfer (FRET) by the oxidization product of p-phenylenediamine (PPD), a chromogenic substrate of horseradish peroxidase (HRP). This approach showed enhanced accuracy and sensitivity with relatively low limit of detection (LOD) of 6.16 × 102 CFU mL-1. Moreover, apple juice contaminated with 1 CFU mL-1 of A. acidoterrestris can be identified after 24 h enrichment. This fluorescence immunoassay could serve as a powerful tool for laboratory identification and on-site inspection of A. acidoterrestris, reducing the adverse effect on the quality of fruit juice.

[Modified Bazhengsan for urination symptoms of chronic prostatis with damp-heat accumulated in the lower jiao].

OBJECTIVE: To study the clinical effects of the modified formula of Bazhengsan on the urination symptoms of chronic prostatis with damp-heat accumulated in the lower jiao. METHODS: A total of 72 chronic prostatitis patients with damp-heat accumulated in the lower jiao were equally randomized to receive the modified formula of Bazhengsan (the trial group) and the primal Bazhengsan (the control group), both for a course of 28 days. Then we observed the changes in the NIH-CPSI scores, scores on Traditional Chinese Medicine (TCM) syndrome, maximum flow rate of urine (MFR) and results of the expressed prostatic secretion test (EPS). RESULTS: Among the 31 patients of the trial group, 6 were cured, 10 achieved excellent results, 9 got improved, and 6 failed to respond, with a total efficacy rate of 80.6%. While in the control group, 4 were cured, 10 achieved excellent results, 11 got improved, and 7 failed to respond, with a total efficacy rate of 78.1%. Significant improvement was observed in NIH-CPSI scores, TCM syndrome and EPS results in both of the groups (P < 0.05), and the trial group showed significant difference from the control in the improvement of urination symptoms (P < 0.05). CONCLUSION: Modified Bazhengsan is effective in the treatment of chronic prostatis with damp-heat in the lower jiao. It can significantly improve its clinical symptoms, especially urination symptoms.

Enhanced biohydrogen production from cotton stalk hydrolysate of Enterobacter cloacae WL1318 by overexpression of the formate hydrogen lyase activator gene.

BACKGROUND: Biohydrogen production from lignocellulose has become an important hydrogen production method due to its diversity, renewability, and cheapness. Overexpression of the formate hydrogen lyase activator (fhlA) gene is a promising tactic for enhancement of hydrogen production in facultative anaerobic Enterobacter. As a species of Enterobacter, Enterobacter cloacae was reported as a highly efficient hydrogen-producing bacterium. However, little work has been reported in terms of cloning and expressing the fhlA gene in E. cloacae for lignocellulose-based hydrogen production. RESULTS: In this study, the formate hydrogen lyase activator (fhlA) gene was cloned and overexpressed in Enterobacter cloacae WL1318. We found that the recombinant strain significantly enhanced cumulative hydrogen production by 188% following fermentation of cotton stalk hydrolysate for 24 h, and maintained improved production above 30% throughout the fermentation process compared to the wild strain. Accordingly, overexpression of the fhlA gene resulted in an enhanced hydrogen production potential (P) and maximum hydrogen production rate (R m), as well as a shortened lag phase time (λ) for the recombinant strain. Additionally, the recombinant strain also displayed improved glucose (12%) and xylose (3.4%) consumption and hydrogen yield Y(H2/S) (37.0%) compared to the wild strain. Moreover, the metabolites and specific enzyme profiles demonstrated that reduced flux in the competitive branch, including succinic, acetic, and lactic acids, and ethanol generation, coupled with increased flux in the pyruvate node and formate splitting branch, benefited hydrogen synthesis. CONCLUSIONS: The results conclusively prove that overexpression of fhlA gene in E. cloacae WL1318 can effectively enhance the hydrogen production from cotton stalk hydrolysate, and reduce the metabolic flux in the competitive branch. It is the first attempt to engineer the fhlA gene in the hydrogen-producing bacterium E. cloacae. This work provides a highly efficient engineered bacterium for biohydrogen production from fermentation of lignocellulosic hydrolysate in the future.