To explore the new method of discriminating Cistanche deserticola, Cynomorium songaricum and Orobanche pycnostachya by using PCR amplification of specific alleles. 30 samples of the different C. deserticola, 21 samples of C. songaricum and O. pycnostachya were collected. The total DNA of the samples were extracted, the ITS sequences from C. deserticola, C. songaricum and O. pycnostachya were amplified by PCR and sequenced unidirectionally. These sequences were aligned by using ClustulW. Specific primer was designed according to the ITS sequences of specific alleles, and PCR reaction system was optimized. Additionally, compare with the identification of specific PCR method and DNA sequence analysis method. The result showed that the 331 bp identification band for C. deserticola and the adulterants not amplified bands by a single PCR reaction, which showed good identification ability to the three species. PCR amplification of specific alleles can be used to identify C. deserticola, C. songaricum and O. pycnostachya successfully.
Alleles ; Cistanche ; classification ; genetics ; DNA Primers ; genetics ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Drug Contamination ; prevention & control ; Phylogeny ; Polymerase Chain Reaction ; methods

Brain Abscess ; diagnosis ; etiology ; therapy ; Cerebellar Diseases ; diagnosis ; etiology ; therapy ; Ear Diseases ; complications ; Humans ; Male ; Young Adult

Objective To evaluate the predictive value of serum C-reactive protein (CRP) for hospital death events in patients with severe community-acquired pneumonia (CAP). Methods The clinical data of 202 patients with CAP in Changhai Hospital and Second People's Hospital of Wuxi between Sep. 2006 and Sep. 2010 were retrospectively reviewed. The clinical and laboratory parameters, including the serum CRP level, white blood cell count, erythrocyte sedimentation rate (ESR) and serum creatine concentration were collected from Hospital Information System (HIS) and Laboratory Information System (LIS). The patients were divided into two groups according to the final death (CAP related complications) or survival of patients in the hospital. The receiver operating curve (ROC) analysis and multivariable logistical model were used to assess the predictive value of CRP on hospital death events. Results The median (interquartile range) serum CRP level of survival patients and patients who died during the hospital stay were 167. 00(132. 50,208. 50) mg/L and 327. 00(246. 25, 411. 50) mg/ L, respectively (Z= - 7. 481,P<0. 001). ROC analysis showed that CRP was an effective predictor for hospital death of CAP patients, with the area under curve (AUC) being 0. 85 (95%CI: 0. 78-0. 91). The optimal cot-off value for serum CRP was 230. 50 mg/L, with the sensitivity being 0. 83(95%CI: 0. 76-0. 89) and specificity being 0. 79(95%CI: 0. 65-0. 89). Logistic regression analysis showed that, after adjusted for age, serum creatinine and ESR, CRP on admission was still independently associated with hospital death of CAP patients (.OR = 13. 42, P<0. 01). Conclusion Increased CRP is an independent risk factor for hospital death events in patients with severe CAP.

Brain ; abnormalities ; pathology ; surgery ; Child, Preschool ; Epilepsy ; etiology ; Humans ; Magnetic Resonance Imaging ; Male

Animals ; Bone Marrow Cells ; cytology ; Brain ; cytology ; physiopathology ; Brain Injuries ; physiopathology ; therapy ; Cell Movement ; Cells, Cultured ; Cisterna Magna ; Immunohistochemistry ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; transplantation

OBJECTIVETo explore the effect of Zengjing Capsule No. 1 (ZJC1) on morphology and motility of sperm in patients with oligospermia (OSM).

METHODSSeventy-two OSM patients were assigned to 2 groups by a randomizing digital table, the treated group and the control group, they were treated respectively by ZJC1 and Wuzi Yanzong Pill (WYP). The changes of density, motility and morphology of sperm in patients before and after 3-month treatment were examined using computerized WLJY-9000 colour semen analysis system with refined Papanicolaou's stain.

RESULTSThe density, motility and morphology of sperm were improved and sperm deformity rate was significantly decreased after treatment in both groups (P < 0.01), but the effects in the treated group were better than those in the control group (P < 0.01).

CONCLUSIONZJC1 can enhance the density and motility of sperm and reduce the sperm deformity rate in patients with OSM.

Adult ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Male ; Oligospermia ; drug therapy ; physiopathology ; Phytotherapy ; Semen Analysis ; Sperm Count ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects ; Young Adult

Signal transducer and activator of transcription (STAT) 3 is a critical transcription factor for cell proliferation and survival. It is activated within cells by many cytokines to mediate immune and inflammatory responses to injury. Inflammatory bowel disease (IBD), represented by Crohn′s disease (CD) and ulcerative colitis (UC), is a chronic inflammatory disease of the intestinal tract. STAT3 has been shown to be abnormally activated in IBD colon tissues by many pro-inflammatory cytokines, leading to disruption of the intestinal mucosal barrier and excessive innate immune and Th17 responses. The persistent chronic inflammation eventually leads to intestinal fibrosis and stenosis. In addition to immune responses, STAT3 is also involved in intestinal fibrosis in IBD by promoting the transcription of fibrosis-related genes. Colitis-associated cancer (CAC) is a particularly aggressive subtype of colorectal cancer and is associated with chronic inflammation-induced IBD. STAT3 has also been associated with CAC initiation and development. STAT3 is overactivated in tumors, which leads to suppression of the anti-tumor activity of immune cells and promotion of cancer cell proliferation, tumor angiogenesis, invasion, and migration. In the present article, we summarize the role of STAT3 in IBD and CAC and the research progress of the related drugs developed for UC and CAC treatment.

OBJECTIVETo evaluate the feasibility and safety of the peripherally inserted central catheters (PICC) as a venous access for newborns who need a long-term venous transfusion.

METHODSSixty-five newborns receiving PICC and 80 newborns receiving peripheral intravenous catheters (PIV) from April 2006 to February 2008 were included in this study. A retrospective cohort study was used to compare the indwelling time of catheters, catheter-related mechanical complications, the incidence of sepsis, and the mortality between the two groups.

RESULTSThe indwelling time of catheters in the PICC and the PIV groups was 18.75+/-7.62 days (range:7-62 days) and 1.49+/-0.57 days (range: 30 minutes to 4 days) respectively. The indwelling time of catheters in the PICC group was significantly longer than that in the PIV group (<0.01). The incidence of catheter-related mechanical complications in the PICC group was significantly lower than that in the PIV group (27.7% vs 63.8%; <0.01). There were no significant differences in the incidence of sepsis and the mortality between the two groups.

CONCLUSIONSThe application of PICC can cause a decrease in the number of venous puncture. PICC is a safe and effective venous access in newborns.

Catheterization, Central Venous ; adverse effects ; Catheterization, Peripheral ; adverse effects ; Humans ; Infant, Newborn ; Sepsis ; etiology ; Time Factors

To study the effect of micro (mi)RNA on cellular proliferation induced by hepatitis B x protein, HBx, in human liver cells and to investigate the underlying molecular mechanism of this cancer-related effect. The human L02 hepatocyte cell line was stably transfected with HBx (L02/HBx) or an HBx mutant (L02/HBx-d382) that induces higher levels of cellular proliferation. The differential miRNA expression profiles were determined by microarray analysis and confirmed by real-time PCR. Two miRNAs, miR-338-3p and miR-551b, that were found to be significantly down-regulated in the L02/HBx-d382 cells were selected for further study and transfected individually into cells using the lipofectamine procedure. The cell survival rate was analyzed by MTT assay, and cell cycles were assessed by flow cytometry. Expressions of cyclinD1, cyclinG1, and E2F1 were assessed by real-time PCR and Western blotting. Compared with the microarray miRNA profile of L02/pcDNA3.0 cells, six miRNAs were up-regulated and five miRNAs were down-regulated in the L02/HBx-d382 cells, while four miRNAs were up-regulated and 12 were down-regulated in the L02/HBx cells. The microarray results were consistent with real-time PCR results. Transfection of miR-338-3p and miR-551b significantly inhibited the cell survival rates (P less than 0.001) and induced G0/G1 phase cycle arrest. According to MTT results: for L02/HBx-d382 cells, compared with lipofectamine or non-transfected (NC) controls, the t value of miR-338-3p was 10.402, 9.133 and the t value of miR-551b was 8.763, 7.403; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 9.105, 8.074 and the t value of miR-551b was 7.673, 7.52. According to flow cytometry results: for L02/HBx-d382 cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 12.173, 11.107 and the t value of miR-551b was 15.364, 13.377; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 15.416, 13.378, and the t value of miR-551b was 13.276, 13.109. The protein levels of cyclinD1, cyclinG1, and E2F1 were significantly reduced by both miR-338-3p and miR-551b ( P less than 0.001). For L02/HBx-d382 cells, compared with lipofectamine or NC controls: E2F1 had t = 11.132, 10.031 and 12.017, 10.973, respectively; cyclinD1 had t = 15.654, 15.013 and 15.447, 14.733, respectively; cyclinG1 had t = 8.017, 7.661 and 7.402, 7.417, respectively. For L02/HBx cells, compared with lipofectamine or NC controls: E2F1 had t = 14.244, 13.331 and 15.022, 14.468, respectively; cyclinD1 had t = 8.695, 8.137 and 7.877, 7.503, respectively; cyclinG1 had t = 7.73, 7.471 and 7.596, 7.41, respectively. In contrast, the mRNA levels for E2F1, cyclinD1, and cylcinG1 showed no significant differences between the miRNA transfected cells and controls. Wild-type HBx and the high proliferation-inducing mutant HBx can influence the miRNA expression profile of L02 cells. HBx down-regulates miR-338-3p and miR-551b in L02 cells, and the high proliferation-inducing mutant has a more robust effect. The mechanism of miR-338-3p- or miR-551b-mediated cell growth inhibition appears to be related to the direct modulation of cyclinD1, cyclinG1, and E2F1.
Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Cycle ; Cell Line ; Cell Proliferation ; Cyclins ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Genes, Viral ; Hepatitis B virus ; genetics ; metabolism ; Hepatocytes ; metabolism ; pathology ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; genetics ; metabolism ; Mutation ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; Real-Time Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; Transfection

BACKGROUNDCytosine deaminase (CD) converts 5-fluorocytosine (5-FC) to 5-fluorouracil (5-FU) in CD/5-FC gene therapy, 5-FU will be mostly converted into nontoxic beta-alanine without uracil phosphoribosyltransferase (UPRT). UPRT catalyzes the conversion of 5-FU to 5-fluorouridine monophosphate, which directly kills CD::UPRT-expressing cells and surrounding cells via the bystander effect. But the pharmacokinetics and the bystander effect of CD::UPRT/5-FC has not been verified in vivo and in vitro. Before the CD::UPRT/5-FC bi-gene therapy system is used in clinical trial, it is essential to monitor the transgene expression and function in vivo. Thus, we developed a preclinical tumor model to investigate the feasibility of using (19)F-magnetic resonance spectroscopy ((19)F-MRS) and optical imaging to measure non-invasive CD and UPRT expression and its bystander effect.

METHODSC6 and C6-CD::UPRT cells were cultured with 5-FC. The medium, cells and their mixture were analyzed by (19)F-MRS. Rats with intracranial xenografted encephalic C6-CD::UPRT glioma were injected intraperitoneally with 5-FC and their (19)F-MRS spectra recorded. Then the pharmacokinetics of 5-FC was proved. Mixtures of C6 and C6-CD::UPRT cells at different ratios were cultured with 5-FC and the cytotoxic efficacy and survival rate of cells recorded. To determine the mechanism of the bystander effect, the culture media from cell comprising 25% and 75% C6-CD::UPRT cells were examined by (19)F-MRS. A comparative study of mean was performed using analysis of variance (ANOVA).

RESULTS(19)F-MRS on samples from C6-CD::UPRT cells cultured with 5-FC showed three broad resonance signals corresponding to 5-FC, 5-FU and fluorinated nucleotides (F-Nuctd). For the C6 mixture, only the 5-FC peak was detected. In vivo serial (19)F-MRS spectra showed a strong 5-FC peak and a weak 5-FU peak at 20 minutes after 5-FC injection. The 5-FU concentration reached a maximum at about 50 minutes. The F-Nuctd signal appeared after about 1 hour, reached a maximum at around 160 minutes, and was detectable for several hours. At a 10% ratio of C6-CD::UPRT cells, the survival rate was (79.55 +/- 0.88)% (P < 0.01). As the C6-CD::UPRT ratio increased, the survival rate of the cells decreased. (19)F-MRS showed that the signals for 5-FU and F-Nuctd in the culture medium increased as the ratio of C6-CD::UPRT in the mixture increased.

CONCLUSIONS(19)F-MRS studies indicated that C6-CD::UPRT cells could effectively express CD and UPRT enzymes. The CD::UPRT/5-FC system showed an obvious bystander effect. This study demonstrated that CD::UPRT/5-FC gene therapy is suitable for 5-FC to F-Nuctd metabolism; and (19)F-MRS can monitor transferred CD::UPRT gene expression and catalysis of substrates noninvasively, dynamically and quantitatively.

Animals ; Antimetabolites ; pharmacokinetics ; therapeutic use ; Cell Line ; Cytosine Deaminase ; genetics ; physiology ; Flucytosine ; pharmacokinetics ; therapeutic use ; Genetic Therapy ; methods ; Glioma ; drug therapy ; therapy ; Humans ; Magnetic Resonance Imaging ; Male ; Pentosyltransferases ; genetics ; physiology ; Rats ; Rats, Sprague-Dawley