Objective: To clone the full-length cDNA of the uridine diphosphate glycosyltransferase (UGT) gene in Bupleurum chinense (BcUGT8), which may be involved with the saikosaponin biosynthesis, and to construct the prokaryotic expression vector. The work will provide the foundation for its further function verification by in vitro expression and activity analysis of the purified protein. Methods: RACE and LD-PCR were used to clone the full-length cDNA of BcUGT8, on the basis of its partial cDNA sequence obtained from our previous high-flux sequencing by Roche (454) GS FLX system. The open reading form (ORF) was PCR cloned using primers with corresponding restriction enzymes cutting sites. The PCR products were digested with corresponding restriction enzymes and then were inserted in expression vector pET-28a (+) to construct the recombinant expression vectors. Results: The full-length cDNA of UGT gene was cloned from B. chinense, and the prokaryotic expression vector was obtained. Conclusion: The full-length cDNA cloning, sequence analysis, and prokaryotic expression vector construction provide a substantial foundation for follow-up bio-function analysis of BcUGT8 through protein expression, purification, and activity analysis in vitro.

ObjectiveTo evaluate the efficacy of self-management to improve the lower urinary tract symptoms (LUTS) and life quality in benign prostatic hyperplasia (BPH) patients.MethodsTwo hundred and twenty-two men were recruited to take αblockers for at least 3 months from March 2008 to September 2009. Participants were randomized to either attend a self-management program in addition to standard care or to standard care alone. Difference of scores of International Prostate Symptom Score (IPSS) and BPH-specific quality of life scale between the two groups was analyzed during enrollment period, the 1st week, the 3rd month and 6th month of therapy.ResultsAll participants were followed up for 6 months.The IPSS scores of the SMI group were 20.5 and 20.5 and the QOL were 50.9 and 50.8 at the 1st week.While the numbers were 19.6 and 19.3 for IPSS and 51.1 and 51.1 for QOL in the control group. There was no statistic difference in the control group.Whereas during the 3rd and 6th month assessment, the scores of IPSS and quality of life of self-management interventional group were statistically significantly lower than those of the control group.ConclusionsSelf-management intervention could reduce the LUTS symptoms and improve quality of life in BPH patients who were taking medicines.

The tissue-specific and MeJA-induced transcriptional levels of BcUGT3 and BcUGT6 in Bupleurum chinense were analyzed in the present study. The transcriptional levels of BcUGT3 in root, leaf, flower and fruit were similar and they all were higher than those in stem. The transcriptional level of BcUGT6 was the highest in leaf and the lowest in flower among in all tested tissues. With non-treated adventitious roots as control, BcUGT6's transcriptional levels were elevated to nearly 2 folds for 2 h, 8 h, 24 h, 2 d and 4 d in MeJA-treated adventitious roots of B. chinense. It showed that the transcriptional level of BcUGT6 was slightly affected by MeJA. While, BcUGT3's transcriptional levels were gradually elevated, and till 4 d after MeJA treatment, the expression level was about 7 folds than that of non-treated control. Using pET-28a (+), the expressions of two genes was investigated. Induced by IPTG, the target proteins were expressed in E. coli and then purified. All the results obtained in the present study will be helpful for follow-up bio-function analysis of BcUGT3 and BcUGT6.
Acetates ; pharmacology ; Bupleurum ; cytology ; enzymology ; genetics ; Cell Membrane ; metabolism ; Cyclopentanes ; pharmacology ; Escherichia coli ; genetics ; Gene Expression ; Gene Expression Regulation, Plant ; drug effects ; Hexosyltransferases ; chemistry ; genetics ; isolation & purification ; metabolism ; Intracellular Space ; metabolism ; Oxylipins ; pharmacology ; Protein Sorting Signals ; Protein Structure, Secondary ; Protein Transport ; Sequence Analysis ; Transcription, Genetic ; drug effects

OBJECTIVE: To study the how's for pharmacists at inpatient pharmacy to provide pharmaceutical care for improved safe and effective use of drugs. METHODS: Pharmacists at inpatient pharmacy can provide pharmaceutical care as per specific clinical needs, and start from such details as preparing the booklet of commonly-used chemotherapy drugs, offering face-to-face advises on drug use for diabetics, and helping nurses to manage the medicine cabinets. RESULTS & CONCLUSION: Pharmacists at inpatient pharmacy should provide the right pharmaceutical care that can meet actual clinical needs.

OBJECTIVETo observe the long-term apparent and structural change of the neo-urethra inner wall reconstructed with different kinds of tissue. To compare the difference between normal and reconstructed urethra for urethra reconstruction with more appropriate materials.

METHODSThe neo-urethra inner wall was observed during operation, through urography and urethroscope. The tissue section of neo-urethra inner wall was observed through light microscope and electron microscope.

RESULTSWith unchanged structure, the appearance of neo-urethra reconstructed with skin or mucosa could be close to normal urethra after a long time. Neo-urethra made of buccal mucosa possesses some of the most important basic structures for normal urethral microenvironment to form. The structure of neo-urethra made of bladder could be close to normal urethra after a long time.

CONCLUSIONSBuccal and bladder mucosa may be a better material than skin for urethra reconstruction.

Adolescent ; Adult ; Biocompatible Materials ; Child ; Female ; Graft Survival ; Humans ; Hypospadias ; surgery ; Male ; Mouth Mucosa ; transplantation ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Tissue Culture Techniques ; Tissue Engineering ; Tissue Transplantation ; methods ; Urethra ; surgery ; Young Adult

OBJECTIVETo understand the projects completion and research progresses in pediatrics which were funded by the National Natural Science Foundation of China (NSFC), and evaluate the accomplishment objectively and justly.

METHODSThe completion status of projects in pediatrics funded by department of clinical medicine II from 2002 to 2006 was analysed retrospectively, and important research achievement and outstanding development in some projects were reported.

RESULTSDuring the period between 2002 and 2006, 420 articles were published, and the average was 8.1 papers per project, which included 56 papers that were published in journals indexed by SCI (the average was 1.1 papers per project). The completion of general project was better than that of "the Young Researchers Fund" and small grant project. Ten post-doctors, 102 doctors and 109 masters were trained. Two projects were awarded with the first grade prize and another 2 with the second grade prize at the provincial and ministerial level, 4 items applied for patent and 1 was granted. These completed projects, which were mainly related to 7 of 12 subspecialties in the field of pediatrics, such as the respiratory disease, nephrology, neurology, cardiology, endocrinology, hematology, neonatology, are the major portion of the application projects and subsidized projects funded by NSFC, and achieved great research progresses.

CONCLUSIONDuring the period between 2002 and 2006, the 52 completed projects in pediatrics showed difference in the distribution and quality of accomplishment among subspecialties and among types of supported projects; there are some gaps between pediatrics and some other clinical basic subspecialties II, this situation released the research status and problems in development of pediatrics in China. The general projects completion was good, and many projects obtained research achievements, which reflect the leading function of NSFC in pediatric research.

Foundations ; Pediatrics ; Periodicals as Topic ; economics ; Research ; economics

OBJECTIVETo establish a method for the expression of glycogen synthase kinase 3β with high purity and biological activity.

METHODSE.coli expression system and baculovirus-insect cell expression system were used to produce the kinase, followed by purification using His-tag and GST-tag and determination of its purity and activity by SDS-PAGE and kinase reaction, respectively.

RESULTSGlycogen synthase kinase 3β produced from E.coli represented 54% of the total bacterial protein, as compared with 96% of the total protein from the insect cell system .Glycogen synthase kinase 3β produced from insect cell exhibited an one-fold higher biological activity than the protein obtained from E.coli.

CONCLUSIONSCompared with the protein from E.coli system, glycogen synthase kinase 3β from the insect cell expression system is endowed with a higher purity and bioactivity.

Animals ; Baculoviridae ; metabolism ; Escherichia coli ; metabolism ; Genetic Vectors ; Glycogen Synthase Kinase 3 ; biosynthesis ; isolation & purification ; Insecta ; cytology

OBJECTIVETo explore the surgical approaches, methods and techniques of repair of cerebrospinal fluid (CSF) rhinorrhea via frontal sinus under transnasal endoscopy.

METHODSCerebrospinal fluid rhinorrhea, located at the posterior wall of the frontal sinus (n = 9) and posterior lateral wall of the frontal recess (n = 4) had been repaired surgically. A transnasal endoscopic approach was chosen in 12 patients and combined approach was used in 1 patient during the first procedure. Three patients needed the second surgery. Among them, one patient needed to repair CSF rhinorrhea, 1 patient needed to treat intracranial abscess of frontal lobe via combined approach and another one was treated because of the complication of frontal cyst.

RESULTSTwelve patients were successfully repaired in the first surgery. Only 1 patient needed second surgery. Two patients occurred complications. One was intracranial infection after surgery, external drainage and packing in the frontal sinus was used. Another was obstructive cyst in frontal sinus, transnasal endoscopic frontal sinusotomy was performed.

CONCLUSIONSCSF rhinorrhea which located at the posterior wall of the frontal sinus can be successfully repaired via transnasal endoscopic approach if the leak was visible under endoscopy. The size of the frontal ostium and leak vantage should be considered to prevent the drainage of the frontal sinus which would result in obstructive cyst in frontal sinus, frontal sinusitis and intracranial infection. Combined approach was suggested to the patients that leakage could not be seen in frontal sinus or frontal ostium was difficult to enlarge.

Adolescent ; Adult ; Cerebrospinal Fluid Rhinorrhea ; surgery ; Child ; Endoscopy ; methods ; Female ; Frontal Sinus ; surgery ; Humans ; Male ; Middle Aged ; Young Adult

Objective:To investigate the effect of Yingyang Gongji Wan (YYGJ) on hepatoma cell line HepG2, and provide evidence for clinical application. Method:YYGJ-contained rats serum was prepared. Then the inhibiory rate of cells was detected by methye thiazolye telrazlium (MTS) method in both YYGJ group and blank group. Apoptosis of HepG2 was detected by TdT-mediated dUT nick-end labeling (TUNEL) method in blank group,YYGJ group, and 5-fluorouracil (5-FU) group. The mRNA expression and protein expression levels of E-cadherin, Vimentin, metalloproteinase-2 (MMP-2) and Smad4 were detected by Real-time quantitative PCR (Real-time PCR) and Western blot respectively. The invasion ability of HepG2 cells was detected by cell migration assay (transwell). Result:YYGJ-contained serum inhibited the proliferation of HepG2 cells in a time and concentration-dependent manner. As compared with blank group, the inhibitory rate was increased in 5%, 10%, and 20% YYGJ-contained serum groups on the third day (P<0.05), and apoptosis rate was also increased significantly in YYGJ and 5-FU groups (P<0.05), but there was no significant difference between 50% YYGJ group and 5-FU group in apoptosis rate. As compared with blank group, the mRNA and protein expression levels of E-cadherin and Smad4 were increased significantly, while Vimentin and MMP-2 mRNA and protein expression levels were decreased significantly in YYGJ and 5-FU groups (P<0.05). HepG2 cell invasive ability was decreased significantly in YYGJ and 5-FU groups as compared to blank group (P<0.05), but there was no significant difference between 50% YYGJ group and 5-FU group. Conclusion:YYGJ-contained serum can inhibit the proliferation of HepG2 cells, induce apoptosis, regulate epithelial-mesenchymal transition (EMT)-related E-cadherin, Vimentin, MMP-2 and Smad4 genes and proteins, and decrease tumor invasion ability. The effect was similar to that of 5-fluorouracil. As a unique prescription, YYGJ can be used as a representative for the treatment of coldness and dampness syndrome of primary liver cancer and its anti-cancer mechanism should be further studied.

OBJECTIVETo explore the expression and clinical significance of Hedgehog signaling transcription factor Gli1 in acute lymphoblastic leukemia (ALL) patients.

METHODSThe clinical specimens were obtained from 32 newly diagnosed and 6 relapsed ALL patients. Normal bone marrow cells from 15 healthy donors were used as controls. Real-time qPCR and Western blot were applied to detect Gli1 mRNA and protein expression in bone marrow mononuclear cells (BMMNC) of these samples respectively. The relation of Gli1 mRNA levels with clinical parameter was also evaluated.

RESULTSThe expression level of Gli1 mRNA in de novo and relapsed ALL patients was significantly higher than that in the normal controls (P < 0.05). There was no stalistically significant difference of the Gli1 mRNA expression between de novo and relapsed ALL cases (P > 0.05). In 24 de novo ALL patients with complete remission (CR) after induction chemotherapy, the levels of Gli1 mRNA were significantly reduced as compared with levels before treatment (P < 0.05). However, in 4 ALL patients without remission, no obvious difference of Gli1 mRNA levels were observed as compared with levels of Gli1 before treatment (P > 0.05). A positive correlation between the Gli1 mRNA expression level and white blood cell count (WBC) was found in the BMMNC of ALL patients (R = 0.725, P < 0.05). Similarly, Gli1 protein expression was significantly higher in the de novo and relapsed ALL cases compared with normal controls. The Gli1 protein level was down-regulated when the ALL patients was in CR.

CONCLUSIONThe expression of Gli1 mRNA and protein has been found to be high in de novo and relapsed ALL patients, and the change of Gli1 expression maybe relate to therapeutic efficacy and prognosis of ALL patients.

Bone Marrow Cells ; Humans ; Induction Chemotherapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Prognosis ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Remission Induction ; Transcription Factors ; Zinc Finger Protein GLI1