OBJECTIVETo observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD.

METHODSTotally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot.

RESULTSThe UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01).

CONCLUSIONZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

Animals ; Calcium Channels ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Infertility ; Male ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Semen Analysis ; Sperm Motility ; Spermatozoa ; TRPV Cation Channels ; metabolism ; Testis ; Ureaplasma Infections ; Ureaplasma urealyticum

Objective To study the clinical efficacy of quadruple therapy for type 2 diabetic mellitus (DM) patients with Helicobacter pylori (Hp) positive peptic ulcer.Methods A total of 176 patients with Hp-positive peptic ulcer were enrolled in the control group and 122 Hp-positive peptic ulcer patients with type 2 diabetes were involved in the treatment group.Both groups were treated with oral rabeprazole 10 mg qd + oral clarithromycin 0.25 g bid + oral amoxicillin 0.5 g bid + oral citrate bismuth potassium 0.3 g qid.Clarithromycin and amoxicillin were used for the first 10 days,the remaining drugs are taking 4 weeks.The clinical efficacy and the incidence of adverse drug reactions between the groups were compared,and the relationship between the type 2 diabetes and the eradication of Hp were analyzed.Results After treatment,the total effective rates of the treatment group and the control group were 86.06％ (105/122 cases) and 94.32％ (166/176 cases) respectively,with statistically significant difference (P ＜ 0.05).The eradication rates of Hp in the treatment group and the control group were 72.13％ (88/122 cases) and 86.93％ (153/176 cases) respectively,and the difference was statistically significant (P ＜0.05).Adverse reactions in the treatment group were nausea,diarrhea and melena;adverse effects in the control group were nausea,diarrhea and vomiting.The incidences of adverse drug reactions in the treatment group and the control group were 9.83％ (12/122 cases) and 10.23％ (18/176 cases),without statistically significant difference (P ＞0.05).Conclusion The clinical efficacy of the control group was better than that of the treatment group,indicating that type 2 diabetes should be considered as a key factor in the treatment of Hp positive peptic ulcer.

Objective To assess the hygienic status of the central air conditioning ventilation system in public places by fuzzy comprehensive evaluation method.Methods According to the comprehensive evaluation method of public place (WS /T 199—2001),a fuzzy comprehensive evaluation method was applied to evaluate central air conditioning ventilation systems in 15 large public places of Jinhua City.Results The qualified rate of central air ventilation system was 53.30%, and the qualified rates of total number of bacteria,the fungi,β-hemolytic streptococcus in air blow and Legionella bacteria in the cooling water all reached 100.00%.At the same time,the qualified rate of bacteria,fungi and dust volume in internal surface of pipe were 93.33%,86.67% and 80.00% respectively.The qualified rate of PM10 was 86.67%. Among 15 central air conditioning ventilation systems,13 systems were evaluated as level Ⅰ,and the other two was as level Ⅱand Ⅳ respectively.TAAlso,14 evaluated units got the fuzzy comprehensive indexes of central air ventilation less than 0.5.Conclusion The hygiene status of central air conditioning ventilation system in Jinhua is acceptable,and the management of hygiene quality should be strengthened.

OBJECTIVESTo study the correlation between positive rates of RNA in clinical confirmed severe acute respiratory syndrome (SARS) patients and its appearance in relation to the development of the disease in order to provide scientific basis for early diagnosis, effective prevention and treatment of the disease.

METHODSOne-step reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the SARS RNA in the clinical specimens from different courses of the disease. The representative amplicons were then sequenced. Chi-square for trend test was performed to study the correlation between positive rates of RT-PCR and at different periods after the onset of the disease.

RESULTSThe fragments amplified from the sputum specimens of SARS patients were shown to share 100% homology with the published SARS-associated coronavirus. Of the different clinical specimens, positive rate in the stools appeared to be the highest (21.55%). Chi-square for trend test revealed that the positive rates of stools and sputa of SARS patients decreased with the development of the disease (chi(2) for trend = 12.55 and 16.408, P = 0.0004 and P = 0.000 05 respectively).

CONCLUSIONOne-step RT-PCR proved to be an effective method for the detection of SARS-associated coronavirus from clinical specimens. Data as indicated that the positive rates of SARS coronavirus were decreasing in SARS patients along with the disease progression.

Adolescent ; Adult ; Aged ; Chi-Square Distribution ; China ; Disease Progression ; Feces ; virology ; Female ; Humans ; Male ; Middle Aged ; Mucus ; virology ; RNA, Viral ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; Severe Acute Respiratory Syndrome ; pathology ; virology

Phosphatidylinositide 3-kinase (PI3K)/protein kinase B (PKB, Akt) pathway plays a major role in proliferation and survival of many types of cells. The inhibitory effect of LY294002, widely applied as an inhibitor of PI3K, in combination with gemcitabine on proliferation of PANC-1 cells was investigated. The expression of PI3K, phosphorylated Akt (p-Akt) and multidrug-resistance like protein (MRP) in normal pancreas tissues, chronic pancreatitis tissues and pancreatic carcinoma tissues was detected. The effects of LY294002 combined with gemcitabine on proliferation of PANC-1 cells and protein levels of p-Akt and MRP were detected. The results showed that the positive expression rate of PI3K, p-Akt and MRP in pancreatic carcinoma tissues was significantly higher than that in normal pancreas tissues and chronic pancreatitis tissues (P<0.01 and P<0.05 respectively). LY294002 could effectively enhance the inhibitory effect of gemcitabine on proliferation of PANC-1 cells. Furthermore, Western blotting revealed that LY294002 combined with gemcitabine reduced the protein levels of p-Akt and MRP, which contributed to the inhibition of proliferation. It is concluded that LY294002 in combination with gemcitabine may represent an alternative therapy for pancreatic carcinoma.
Adult ; Antimetabolites, Antineoplastic ; administration & dosage ; Cell Proliferation ; drug effects ; Chromones ; administration & dosage ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; Dose-Response Relationship, Drug ; Drug Synergism ; Drug Therapy, Combination ; methods ; Enzyme Inhibitors ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Morpholines ; administration & dosage ; Pancreatic Neoplasms ; drug therapy ; pathology ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; Treatment Outcome ; Tumor Cells, Cultured

This study was aimed to investigate the anti-leukemia activity of Tel03 in vivo. The K562 xenografted leukemia model was established and mice were divided randomly into three groups. Mice of different group were treated with PBS (control), 5 mg/kg Tel03 or 15 mg/kg Tel03 (ip, twice a week) respectively. Tumor volume, body weight and other behavior were observed regularly. Cell apoptosis was detected with TUNEL assay and the expression levels of Bcl-2 and Bax were detected by Western blot. The results indicated that Tel03 exerted anti-leukemia activity in mouse model. Tel03 significantly reduced tumor volume in Tel03-treated group compared with control. In addition, 5 mg/kg Tel03 induced cell apoptosis without exerting apparent toxicity in mice. After Tel03 treatment, the expression of Bcl-2 was inhibited, however, the expression of Bax was up-regulated. It is concluded that G-quadruplex ligand Tel03 can induce cell apoptosis in leukemia mouse model, and this agent may be a potential anticancer drug.
Animals ; Apoptosis ; Female ; G-Quadruplexes ; Humans ; K562 Cells ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; metabolism

Hematopoietic progenitor kinase 1 ( HPK1 ) , also known as MAP4K1 , is a serine/threonine protein kinase and a member of the MAP4K family of mammalian Ste20-related pro¬tein kinases.Recent studies have found that HPK1 is assoeiated with the occurrence and progression of a variety of tumors, and may play an important role in some malignant tumors.This pa¬ per reviews the HPK1 signaling pathway, its relationship with tumor and drug development progress, so as to provide referenee for the research of HPK1 protein kinase.

The purpose of this study is trying to analysis the homology between four lentogenic Class I genotype 3 Newcastle disease virus isolates from different hosts with NDV strain NDV 08-004, which was the first obtained complete genome sequence virus of class I genotype 3. The full-length genome of NDV isolates, JS/3/09/Ch, ZJ/3/10/Ch, AH/2/10/Du and JS/9/08/Go,were determined by RT-PCR and then an alyzed. All the genomes are 15 198 nucleotides (nt) in length. Compared with the full genome sequences of Class II NDV stains (genotype IV-IX),four isolates has a 6-nt deletion in the non-coding region of nuclear phosphoprotein gene between nucleotides 1 640-1 641 and 12-nt insertion in the coding region of phospho protein gene between nucleotides 2 381-2 382. All the isolates have the motifs 112EQ/RQE/GRL117 at the cleavage site of the fusion protein, which is typical of lenogenic NDV strains, and it is in agreement with the result of pathogenic tests. The full-length genome of 4 genotype 3 NDV isolates shared 93% nucleotide identity with NDV08-004. The results of alignment of 6 viral genes showed that NP gene shared the highest identity (98.3%-96.4%) and P gene shared the lowest identity (96.1%-91.9%). The results show the following two points. First, it is concluded that the isolates from different hosts share the same genotype has the insignificant divergence in the genetic information. Second, it is proposed that the mutation rates of NP/F/L genes are lower than P/M/HN genes.
Animals ; Genome, Viral ; genetics ; Genomics ; Genotype ; Host Specificity ; genetics ; Newcastle disease virus ; classification ; genetics ; isolation & purification ; Phylogeny

Objective: To explore the effects of Zhibai Dihuang Decoction (ZDD) on mitochondrial cytochrome oxidase (COX) in the spermatogenic cells of rats with ureaplasma urealyticum (UU) infection. METHODS: From forty 4－5 months old SD rats, 30 were randomly selected for the establishment of the model of testicular UU infection by inoculating the bladder with UU suspension and the other 10 injected with normal saline as controls (group A). At 7 days after inoculation, the rat models of testicular UU infection were treated orally with normal saline (group B), ZDD at 1 g per kg of the body weight per day (group C), and azithromycin at 0.105 g per kg of the body weight per day (group D), respectively, once daily for 21 days. Then all the animals were sacrificed and the epididymal and testicular tissues collected for examination of sperm motility with the color sperm dynamic detection system, measurement of the COX activity with the immunohistochemical DAB method, and determination of the mRNA expressions of COXⅠ and COXⅡ by RT-PCR. RESULTS: Compared with group A, group B showed significant decreases in such sperm parameters as grade a sperm (［1.03 ± 0.09］ vs ［0.07 ± 0.03］ %, P<0.01), grade b sperm (［2.07 ± 0.52］ vs ［0.35 ± 0.13］ %, P<0.01), straight line velocity (VSL) (［10.95 ± 0.98］ vs ［6.78 ± 1.05］ μm/s, P<0.01), curvilinear velocity (VCL) (［42.03 ± 1.35］ vs ［38.10 ± 7.65］ μm/s, P>0.05), average path velocity (VAP) (［16.22 ± 1.52］ vs ［10.05 ± 1.80］ μm/s, P<0.01), and the mRNA expressions of COX Ⅰ (［2.25 ± 0.24］ vs ［0.93 ± 0.10］ %, P<0.01) and Ⅱ (［6.72 ± 0.37］ vs ［2.95 ± 0.78］ %, P<0.01). After treatment, all the parameters were remarkably increased in groups C and D (grade a sperm: ［1.11 ± 0.30］ and ［0.60 ± 0.19］%; grade b sperm: ［2.40 ± 0.59］ and ［1.32 ± 0.27］ %; VSL: ［12.11 ± 1.62］ and ［11.47 ± 1.21］ μm/s; VCL: ［54.30 ± 2.35］ and ［45.75 ± 1.64］ μm/s; VAP ［18.40 ± 1.27］ and ［16.69 ± 1.02］ μm/s; expression of COXⅠ mRNA: ［1.86 ± 0.30］ and ［1.74 ± 0.17］ %) as compared with those in group B (P<0.05or P<0.01) except the COX activity and the expression of COX Ⅱ mRNA (P>0.05), and all the parameters were significantly higher in group C than in D (P<0.05or P<0.01). CONCLUSIONS UU infection can reduce grades a and b sperm, linear, curvilinear and mean sperm velocities, and the mRNA expressions of COX Ⅰ and Ⅱ while ZDD can improve these parameters. The improvement of sperm motility may not be associated with the activity of COX, and the COX activity may be related to the mRNA expression of COX II but not that of COXⅠ.
Animals ; Anti-Bacterial Agents ; therapeutic use ; Azithromycin ; therapeutic use ; Drugs, Chinese Herbal ; pharmacology ; Electron Transport Complex IV ; metabolism ; Epididymis ; drug effects ; enzymology ; Humans ; Male ; Mitochondria ; drug effects ; enzymology ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; Spermatozoa ; drug effects ; enzymology ; physiology ; Ureaplasma Infections ; drug therapy ; enzymology ; Ureaplasma urealyticum

Fungal pathogens represent major problems for human health and agriculture. As eukaryotic organisms, fungi share some important features with mammalian cells. Therefore, current anti-fungal antibiotics often can not distinguish between fungi and mammalian cells, resulting in serious side effects in mammalian cells. Accordingly, there is strong impetus to develop antifungal alternatives that are both safe and effective. The E1 family of colicin are channel-forming bacteriocins produced by Escherichia coli, which are bactericidal only to E. coli and related species. To target the channel-forming domain of colicin to fungal cell membrane, we engineered a sexual mating pheromone of Candida albicans, α-factor pheromone to colicin Ia. A peptide was constructed consisting of an α mating pheromone of C. albicans fused to the channel-forming domain of colicin Ia to create a new fusion protein, pheromonicin-CA (PMC-CA). Indirect immunolabeling showed that the PMC-CA bound to fungal cells and inhibited growth in the laboratory and field. In the field, the protective activity of pheromonicin against rice blast disease was significantly greater, on a molar basis, than that of triazoles, tricyclazole or isoprothiolane. These results suggest that fusion peptides may be of value as fungicidal agents under agricultural conditions.
Candida albicans ; chemistry ; Colicins ; chemistry ; Fungicides, Industrial ; chemistry ; Mating Factor ; Peptides ; chemistry ; Protein Engineering