In this paper, Liuwei Dihuang pill was used to study the identification of Chinese patent medicine by fluorescence sequencing typing technology. The DNA of Paeonia suffruticosa was used as template to amplify by five pair of FAM fluorescence labeling primers. Then, the amplified products were sequenced. The sequencing results were analyzed by GeneMarker V1.80 to screen the best fluorescence labeling primers. As a result, psbA-trnH fluorescence labeling primer was used to identify the raw materials of Liuwei Dihuang pill. The results showed that three kinds of raw plant medicinal materials in Liuwei Dihuang pill were able to be correctly identified by psbA-trnH fluorescence labeling primer. The fluorescence sequencing typing technology can stably and accurately distinguish raw medicinal materials in Chinese patent medicine.
DNA Primers ; chemistry ; genetics ; DNA, Plant ; chemistry ; genetics ; Drugs, Chinese Herbal ; chemistry ; standards ; Fluorescent Dyes ; chemistry ; Plants, Medicinal ; chemistry ; genetics ; Polymerase Chain Reaction ; instrumentation ; methods ; Quality Control ; Staining and Labeling

BACKGROUND: A long-term follow-up indicates that orthotopic ileal neobladder can cause acid-base balance and nutritional metabolic disorder. Otherwise, a long mesenterium is necessary to balance the tension of bladder at pelvic cavity and urinary inosculation due to a high position of ileum. On the contrary, sigmoid neobladder is near by urinary canal, and orthotopic sigmoid neobladder as a bladder substitute after radical cystectomy has few effects on acid-base balance of electrolytes, nutritional metabolism and secretion of mucus.OBJECTIVE: To compare the clinical results of these two operations basis on long-term follow-up. DESIGN: Retrospective analysis.SETTING: Department of Urinary Surgery, the Second Affiliated Hospital of Kunming Medical College.PARTICIPANTS: 164 patients with carcinoma of bladder were selected from Department of Urinary Surgery, the Second Affiliated Hospital of Kunming Medical College form January 1995 to March 2005. Ninety-six of them, including 74 males and 22 females, with age of 43-74 years and the average age of 65 years, accepted the operation of orthotopic ileal neobladder were regarded as the ileal neobladder group, and the other 68, including 64 males and 4 females, with age of 51-72 years and the average age of 62 years, accepted the operation of orthotopic sigmoid neobladder were regarded as the sigmoid neobladder group. All patients were finally diagnosed as pathological examination, and informed consent was provided by all patients. Treatment plan was approved by the local ethical committee.METHODS: ① Orthotopic ileal neobladder: Once the bladder was removed, a segment of ileum about 40-60 cm in length was isolated. In the operative procedure, the distal part of ileum which connected to the caecum often kept, the length of which was 15-20 cm. Both distal ends of the ureters were anastomosed to the homolateral not been split end of the isolated bowel. A perforation was constructed at the bottom of the pouch which served as the outlet, this outlet was then anastomosed to the proximal portion of the remaining urethra. ② Orthotopic sigmoid neobladder: After surgically removing the bladder, a part of the sigmoid colon, the length of which was 30-40 cm was isolated. Other operations were as the same as those mentioned above. MAIN OUTCOME MEASURES: Time of operation, blood loss during the procedure, length of time confined to bed, time of indwelling catheter, the ability to maintain continence and urinate, the results of urodynamic studies, and pouch related complications after operation.RESULTS: In 164 patients, 12 (7.3%) were lost to follow-up. The mean follow-up times were 46 months in the group of orthotopic ileal neobladder and 42 months in the group of orthotopic sigmoid neobladder, respectively. Blood loss during the procedure and the ability to maintain continence and urinate were similar in the two groups (P > 0.05). Compared with sigmoid neobladder group, the ileal neobladder group spent more time on operation, keeping the bed and indwelling catheter. The max volume of ileal pouch was higher than that of sigmoid pouch, and the difference was significant in statistic analysis (t=2.56-3.08, P < 0.05-0.01). Incidence of complication of ileal pouch (16.7%, 29.2%) was higher than that of sigmoid pouch (9%, 16%). The incidence in the early phase was not significantly different, but that in the late phase was significantly different (x2=5.426, P < 0.05).CONCLUSION: Compared with orthotopic ileal neobladder, sigmoid neobladder is worthy of being preferred for its shorter operative time, faster recovery and lower rate of pouch related complications.

Liquid-phase microextraction is a novel pretreatment technique for biological samples developed on the basis of liquid-phase extraction technology, which is simple, rapid, economical, and environmentally friendly, and has been widely used in the analysis of biological matrix samples such as blood, urine, and saliva. In this paper, we review the basic principles of the main modes of liquid-phase microextraction techniques, i.e., single-drop microextraction, dispersive liquid-liquid microextraction, and hollow-fiber liquid-phase microextraction, and the progress of their applications in biological sample pretreatment by reviewing the literature in the past five years, with a view to providing technical support and reference for sample pretreatment in the fields of in vivo drug analysis, pharmacokinetic studies and new drug development.

It is unanimously accepted that stroke is a highly heterogeneous disorder.Different subtypes of ischemic stroke may have different risk factors,clinical features,and prognoses.The aim of this study was to evaluate the risk factors,clinical characteristics,and prognoses of different subtypes of ischemic stroke defined by the Trial of ORG10172 in Acute Stroke Treatment (TOAST) criteria.We prospectively analyzed the data from 530 consecutive patients who were admitted to our hospital with acute ischemic stroke within 7 days of stroke onset during the study period.Standardized data assessment was used and the cause of ischemic stroke was classified according to the TOAST criteria.Patients were followed up till 30 and 90 days after stroke onset.It was found that large-artery atherosclerosis was the most frequent etiology of stroke (37.4％),and showed the highest male preponderance,the highest prevalence of previous transient ischemic attack,and the longest hospital stay among all subtypes.Small artery disease (36.4％) was associated with higher body mass index,higher plasma triglycerides,and lower plasma high-density lipoprotein cholesterol than cardioembolism.Cardioembolism (7.7％),which was particularly common in the elderly (i.e.,individuals aged 65 years and older),showed the highest female preponderance,the highest prevalence of atrial fibrillation,the earliest presentation to hospital after stroke onset,the most severe symptoms on admission,the maximum complications associated with an adverse outcome,and the highest rate of stroke recurrence and mortality.Our results suggest that ischemic stroke should be regarded as a highly heterogeneous disorder.Studies involving risk factors,clinical features,and prognoses of ischemic stroke should differentiate between etiologic stroke subtypes.

Lian Qiao Bai Du Wan was used to study the identification of Chinese patent medicine by molecular marker technique. DNA was extracted through modified CTAB method. The psbA-trnH and rbcL sequences were gradient amplified, and PCR products were ligated with the pEASY-T5 vector and then transformed into Trans1-T1 cells, respectively. Clones were selected randomly and sequenced. All sequences were analyzed by BlastN and the neighbor-joining (NJ) phylogenetic tree was constructed by MEGA 4.0. The results showed that nine kinds of medicinal materials can be identified by psbA-trnH sequences, and six kinds of medicinal materials by rbcL sequences from Lian Qiao Bai Du Wan. Molecular marker technique can stably and accurately distinguish multi-origin medicinal materials in Chinese patent medicine.
Base Sequence ; Chloroplasts ; genetics ; Cluster Analysis ; DNA Barcoding, Taxonomic ; DNA, Chloroplast ; genetics ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Drugs, Chinese Herbal ; chemistry ; Forsythia ; chemistry ; genetics ; Phylogeny ; Plants, Medicinal ; chemistry ; genetics ; Polymerase Chain Reaction ; Ribulose-Bisphosphate Carboxylase ; genetics ; Sequence Analysis, DNA ; Species Specificity

Adult ; Humans ; Kidney Neoplasms ; surgery ; Laparoscopy ; methods ; Male ; Nephrectomy ; methods ; Retroperitoneal Space ; surgery

The bioinformatics software was used to predict the B cell and T cell epitopes of Toxoplasma gondii microneme 16 (TgMIC16) followed by epitopes display on the yeast of Saccharomyces cerevisiae.B and T cell epitopes of TgMIC16 were predicted by DNAStar and IEDB,and software of SYFPEITHI and BIMAS,respectively.According to the predicted results,the antigenic epitope region was expressed with pCTCON2/EBY100 display system.The expression protein was detected by indirect immunofluorescence (IFA) and flow cytometry.Results showed that there were potential B/T cell epitopes in TgMIC16 and concentrated in the aa343-625 region.The epitope region was successfully displayed on the surface of yeast cells,and the optimal induction time was 24 hours.The study would lay the foundation for the development and efficacy evaluation of the yeast carrier vaccine in the further research.

BACKGROUND: The perioperative hemorrhage of knee surgeries is a difficulty in clinic, and the efficacy of tranexamic acid to reduce postoperative bleeding has attracted more attention, but choosing which administrations remains controversial. OBJECTIVE: To investigate the efficacy of tranexamic acid by intravenous injection or articular injection for reducing the perioperative hemorrhage of total knee arthroplasty. METHODS: Sixty patients undergoing unilateral total knee replacement were enrolled, and were then randomized into three groups (n=20 per group): no tranexamic acid administration (group A); intravenous dropping of 15 mg/kg tranexamic acid before tourniquet application plus 10 mg/kg tranexamic acid at 3 hours postoperatively (group B); articular injection of 50 mL saline diluted with 1 g tranexamic acid through a drainage tube (group C). Two-hour closure of drainage tube was performed in all patients. The postoperative dominant and hidden blood loss, blood transfusion rate, pulmonary embolism as well as lower extremity deep venous thrombosis were recorded. RESULTS AND CONCLUSION: (1) The dominant and hidden blood loss in the groups B and C were significantly less than those in the group A (P < 0.05); the dominant blood loss showed no significant difference between groups B and C (P > 0.05); the group B exhibited a significantly less hidden blood loss compared with group C (P < 0.05). (2) The blood transfusion rate in the groups B and C was significantly lower than that in the group A (P < 0.05). (3) No pulmonary embolism or lower extremity deep venous embolism occurred during 3-month follow-up. (4) That is to say, tranexamic acid can obviously reduce perioperative blood loss and blood transfusion rate without pulmonary embolism or lower extremity deep venous thrombosis, and intravenous administration exerts better clinical effectiveness.

Objective To compare the clinical efficacy of orthotopic ileal neobladder versus ortho- topic sigmoid neobladder.Methods The data of 96 patients who had undergone orthotopic ileal neoblad- der and 68 patients who had undergone orthotopic sigmoid neobladder were retrospectively analyzed.The perioperative condition,urinary continence,urodynamics,and pouch-related complications were compared between the 2 groups.Results Of all the 164 patients,12(7.3%)were lost to follow-up.The mean fol- low-up was 46(2-86)months in orthotopic ileal neobladder group,and 42(4-78)months in orthotopic sigmoid neobladder group.There was no significant difference in intraoperative blood loss and postoperative urinary continence between the 2 approaches(P＞0.05).However,compared with sigmoid neobladder group,ileal neobladder group had longer operative time and postoperative recovery time,and got a bigger pouch(P＜0.05).The early and late pouch-related complication rates of ileal neohladder group were 16. 7% and 29.2%,which were higher than those of sigmoid neobladder group.During the follow-up,tumor recurred in 3 cases of ileal neobladder group,but none in sigmoid neobladder group.Conclusions Ortho- topic ileal neobladder and sigmoid neobladder are similar in operative difficulties,and both can achieve satis- factory clinical results.Compared with ileal neobladder,sigmoid neobladder has shorter operative time, quicker recovery and lower rate of pouch-related complications,thus is a preferred procedure.

OBJECTIVETo study the ultrastructural changes of the rat convoluted seminiferous tubule after alcohol consumption.

METHODSForty-eight Wistar mature male rats were divided into two groups randomly: control group (A) and experimental one (B). 6 ml/(kg x d) of 50 degrees alcohol was perfused through the gastric tube for 39 days in Group B; and 6 ml/(kg x d) of normal saline was supplemented in Group A. The ultrastructure of the rat convoluted seminiferous tubule was observed by transmission electron microscope at day 14, 27 and 40.

RESULTSIn Group A, the pykno-basement membrane was unstriated and uniform, Sertoli cells showed cytoplasmic profusion, with big nucleus, well-distributed nucleoplasm, distinct nucleolus, more mitochondria and plain hierarchical tight-junction. And the ultrastructure of the rat convoluted seminiferous tubule in Group B began to change at the end of the first spermatogenic cycle (D 14) and changed more and more evidently with the ethanol administration, mainly as follows: (1) more lysosomes and vacuolisation found in Sertoli cells, and organelles decreased and blurry; (2) more and bigger vacuoles among the spermatogonia, Sertoli cells and basement membrane; (3) obvious apoptosis of spermatogonia and apoptotic bodies aggregated near the membrane; (4) more cytoplasm and vacuolisation in the sperm of the convoluted seminiferous tubule, and disarranged, deleted or clustered mitochondria in the sperm tail; (5) blurry and rigid tight-junction; (6) thickened, wrinkled or broken basement membrane and under-basement

CONCLUSIONAlcohol can cause ultrastructural changes of the basement membrane, tight-junction and Sertoli cells of the membrane. rat convoluted seminiferous tubule and apoptosis of spermatogonia.

Animals ; Apoptosis ; drug effects ; Basement Membrane ; drug effects ; pathology ; Ethanol ; toxicity ; Male ; Microscopy, Electron, Transmission ; Random Allocation ; Rats ; Rats, Wistar ; Seminiferous Tubules ; drug effects ; ultrastructure ; Sertoli Cells ; drug effects ; pathology