The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the conversion of HMG-CoA to mevalonate in mavalonic acid pathway, which is the first committed step for isoprenoid biosynthesis in plants. However, it still remains unclear whether HGMR gene plays a role in the isoprenoid biosynthesis in Dendrobium officinale, an endangered epiphytic orchid species. In the present study, a HMGR encoding gene, designed as DoHMGR1 (GenBank accession JX272632), was identified from D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods, for the first time. The full length cDNA of DoHMGR1 was 2 071 bp in length and encoded a 562-aa protein with a molecular weight of 59.73 kD and an isoelectric point (pI) of 6.18. The deduced DoHMGR1 protein, like other HMGR proteins, constituted four conserved domains (63-561, 147-551, 268-383 and 124-541) and two transmembrane motifs (42-64 and 85-107). Multiple sequence alignment and phylogenetic analyses demonstrated that DoHMGR1 had high identity (67%-89%) to a number of HMGR genes from various plants and was closely related to Vanda hybrid cultivar, rice and maize monocots. Real time quantitative PCR (qPCR) analysis revealed that DoHMGR1 was expressed in the three included organs. The transcripts were the most abundant in the roots with 2.13 fold over that in the leaves, followed by that in the stems with 1.98 fold. Molecular characterization of DoHMGR1 will be useful for further functional elucidation of the gene involving in isoprenoid biosynthesis pathway in D. officinale.
Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Dendrobium ; enzymology ; genetics ; Gene Expression Regulation, Plant ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Molecular Weight ; Phylogeny ; Plant Leaves ; enzymology ; genetics ; Plant Roots ; enzymology ; genetics ; Plant Stems ; enzymology ; genetics ; Plants, Medicinal ; enzymology ; genetics ; Sequence Alignment ; Sequence Homology, Amino Acid

Objective To analyse the follow-up of one family with generalized epilepsies with febrile seizures plus (GEFS +).Methods We conducted a family with GEFS + by sexs,ages, seizure manifestation,electroencephalogram (EEG),and so on.Results There were 36 people in 5 generations of the family in all,including 14 patients(8 cases were male and 16 cases were female).Their ages were from 4 years and 5 months to 8 years.There were 8 cases febrile seizures (FS),4 cases with FS + and 1 case with FS + and absence seizures in 13 patients except 1 case without adequate knowledge.The Results of ECG indicted that 12 cases were normaland 4 cases with FS + and 1 case with absence seizures had epileptic discharges.Conclusions GEFS + is a common kind of inherited epilepsic syndrome and occur in childhood.So it is greatly important for epileptic children to know GEFS +

Objective To investigate the clinical significance of generalized epilepsy with febrile seizures plus(GEFS+ ). Methods The data of one family with GEFS+ were retrospectively analyzed by studying clinical manifestations, physical examinations, electroencephalogram(EEG), 24 hours dynamic EEG monitoring, et al. Some of the patients were examined by CT. Results Ⅳ 12, her chief complaints when admitted to hospital were frequent spasm for 3 days. She began to appear febrile seizures (FS) from 8 months after birth, and frequent generalized tonic - clonic FS appeared during that time. There were 36 people in 5 generations of the family including 14 patients (8 males and 6 females) ,aged from 4 years and 5 months to 82 years. FS presented in 8 cases (Ⅱ 2, Ⅲ1, Ⅲ4, Ⅲ6, Ⅳ1, Ⅳ11, Ⅳ17, Ⅴ2),febrile seizures plus(FS +) in 4 cases ( Ⅳ2, Ⅳ12, Ⅳ13, Ⅳ14), ES + and absence seizures in 1 case ( Ⅴ1 ), uncertain type in 1 case (Ⅰ2). The results of EEG indicated that 12 cases were normal and 4 cases with FS+ and 1 case with absence seizures had epileptic discharges. Apart form Ⅳ13, Ⅳ14 who were treated with magnesium valproate, the dosage for the other patients decreased, or medicine terminated or without medicine, and all the patients had no recurrence of seizures. The intelligence, movement development and neurological examinations of the family were all normal. Head CT scan of 3 cases were normal. Conclusions GEFS+ is autosomal dominant inheritance disease with conspicuous genetic heterogeneity and phenotypic heterogeneity. The apprehension of GEFS+ plays an important role in diagnosis and differential diagnosis of epilepsy in childhood.

Objective To evaluate the efficacy of argon surper-cryosurgery system (ASSS) in the treatment of huge hepatic multi-nidus carcinoma at advanced stage in cases of which conventional surgery is impossible. Methods Twenty-two patients with huge hepatic multi-nidus carcinoma at advanced stage, incurable with conventional surgery, were treated by ASSS in combination with entire or reduced-size excision. In terms of combined treatment, 1 patient received reduced-size operation and portal vein chemotherapeutic pump, 2 with entire excision and portal vein chemotherapeutic pump, 1 with entire excision and liver artery chemotherapeutic pump, 3 with only liver artery chemotherapeutic pump, and 6 with portal vein chemotherapeutic pump. Nine patients were simply treated by ASSS. Results The surgical procedures went smoothly in all cases and the complications, such as bleeding at the puncture aperturae, thorax effusion, bile leakage and liver cataphora, were successfully managed. The follow-up study was performed and all patients surived with the longest survival of 11 months up till now. Conclusions ASSS offers a new treatment method for huge hepatic multi-nidus carcinoma at advanced stage, but the indications must be carefully observed, and the surgical skills and prevention of complication are of great importance.

Objective To evaluate the efficacy of argon surper-cryosurgery system (ASSS) in the treatment of huge hepatic multi-nidus carcinoma at advanced stage in cases of which conventional surgery is impossible. Methods Twenty-two patients with huge hepatic multi-nidus carcinoma at advanced stage, incurable with conventional surgery, were treated by ASSS in combination with entire or reduced-size excision. In terms of combined treatment, 1 patient received reduced-size operation and portal vein chemotherapeutic pump, 2 with entire excision and portal vein chemotherapeutic pump, 1 with entire excision and liver artery chemotherapeutic pump, 3 with only liver artery chemotherapeutic pump, and 6 with portal vein chemotherapeutic pump. Nine patients were simply treated by ASSS. Results The surgical procedures went smoothly in all cases and the complications, such as bleeding at the puncture aperturae, thorax effusion, bile leakage and liver cataphora, were successfully managed. The follow-up study was performed and all patients surived with the longest survival of 11 months up till now. Conclusions ASSS offers a new treatment method for huge hepatic multi-nidus carcinoma at advanced stage, but the indications must be carefully observed, and the surgical skills and prevention of complication are of great importance.

Background A virtual reality simulator provides a novel training model for improving surgical skills in a variety of fields.They can simulate a variety of surgical scenarios to improve the overall skills required for endoscopic operations,and also record the operative process of trainees in real-time and allow for objective evaluation.At present,some simulators for transurethral resection of the prostate (TURP) are available.The utility of virtual reality simulators in training of transurethral prostatectomy was investigated.Methods Thirty-eight urologists were randomly selected to take part in a simulation based training of TURP using the TURPSimTM system.Pre and post-training global rate scale (GRS) scores and objective parameters recorded by the simulator were assessed.Then,questionnaires were filled out.Results Compared with baseline levels,the GRS scores of trainees increased (18.0±4.0 vs.12.4±4.2,P＜0.001),while the rate of capsule resection (26.3％±0.6％ vs.21.2％±0.4％,P ＜0.001),amount of blood loss ((125.8±86.3) ml vs.(83.7±41.6) ml,P ＜0.001),external sphincter injury (3.6±2.9 vs.2.0±2.0,P ＜0.001)decreased significantly after training.Most trainees were satisfied with the simulator based training and believed that the simulator accurately mimicked actual surgical procedures and could help improve their surgical skills.Conclusions As a new method of training on transurethral prostatectomy skills,training of TURP using a virtual simulator can help urologists improve their surgical skills and safety.Therefore,the application of the TURPSimTM system in education and training of urologic surgery is warranted.

OBJECTIVETo probe into a safe and effective method for treatment of functional ventricular premature, and develop a new preparation of Chinese herbs with high science and technique contents.

METHODSTwo hundred and thirty-eight cases were randomly divided into a slowly-releasing medication group (n = 118) and a western medicine group (n = 120). The slowly-control needle group were treated with intramuscular injection of slowly-releasing medication new type preparation made by proved recipe of Chinese herbs with replenishing and strengthening pectoral qi, and nourishing yin and tranquillization into Tanzhong (CV 17), Neiguan (PC 6), Xinshu (BL 15), Pishu (BL 20) and Feishu (BL 13) by a trocar to slowly release and control releasing of the medicine so as to maintain lasting stimulation; the western medicine group with oral administration of Rythmol, thrice daily, 150 mg each time.

RESULTSThe cured rate and the cured and markedly effective rate for the ventricular premature was 51.7% and 71.2% in the slowly-releasing medication group, which were significantly higher than 28.3% and 50.8% in the western medicine group (P < 0.001).

CONCLUSIONThe slowly-releasing medication new type preparation has a better therapeutic effect on functional ventricular premature, with high safety.

Acupuncture Points ; Acupuncture Therapy ; methods ; Adult ; Aged ; Delayed-Action Preparations ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Ventricular Premature Complexes ; physiopathology ; therapy

GLuc (Gaussia luciferase) is a secreted luciferase with high sensitivity. In this study, we primarily compared expression character of PTTR with that of PCMV, relied on easy secretion, high sensitivity and simple and fast detection of GLuc. We firstly constructed two plasmids pAAV2-neo-TTR-GLuc and pAAV2-neo-CMV-GLuc. Then, 4 cell lines were transfected with the two plasmids in aid of Lipofectamine 2000, including Huh7 and HepG2, which are derived from liver cells, as well as HEK293 and HeLaS3 cells, which are non-liver cell lines. We monitored the expression of GLuc in the supernatant of these cell cultures at different time points post-transfection. Furthermore, we injected the two plasmids with different doses into BALB/c mice by the means of hydrodynamic delivery and monitored the GLuc expression in vivo with 2.5 microl tail tip blood since 2 h post-injection. The cell assay results suggested that the expression of GLuc driven by CMV promoter was significantly higher than that of GLuc driven by TTR promoter. And, the luciferase activity of GLuc driven by CMV promoter was 50-300 times higher than that of GLuc driven by TTR promoter in HEK293 and HeLaS3 cell lines, but less than 10 times higher than that of GLuc driven by TTR promoter in the HepG2 and Huh7 cell lines, indicating the relative liver-specificity of TTR promoter. In the animal assay, the higher luciferase activity was determined in CMV promoter group than in TTR promoter group at different doses of the two plasmids. But the expression patterns for the two promoters differed obviously. The expression of GLuc driven by CMV promoter reached the maximum 10 hours post-injection and declined rapidly; while the expression of GLuc driven by TTR promoter reached the maximum 48 hours after delivery, and declined very slowly. These results implied that PTTR could keep expression of driven gene in a long time although its expression intensity is lower than PCMV's. Thus, it is more suitable for maintaining longer expression of target genes in liver.
Animals ; Cell Line ; Cytomegalovirus ; genetics ; metabolism ; Gene Expression ; Gene Transfer Techniques ; Genes, Reporter ; Humans ; Luciferases ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Prealbumin ; genetics ; metabolism ; Promoter Regions, Genetic

BACKGROUNDVirtual reality (VR) has been recognized as a useful modality in the training of surgical skills. With respect to basic endoscopic skill training of urology, we sought to investigate the effectiveness of the UroMentor(TM) virtual reality simulator (VRS) in the skill acquisition of flexible cystoscopy.

METHODSUrologists familiar with rigid cystoscopy procedures were selected to take part in a virtual training course of flexible cystoscopy. Changes in total operating time, frequency of injury, number of digital markers inside the bladder, and the global rating scale (GRS) scores were assessed following eight repeated training sessions on the UroMentor(TM).

RESULTSEighteen urologists voluntarily took part in the study. Total operating time was significantly lower after eight sessions of training by comparison ((111 ± 10) seconds and (511 ± 67) seconds, respectively; P < 0.001). Additionally, the frequency of injury decreased with training from (12 ± 2) times to (5 ± 1) times (P < 0.001), while the number of digital markers observed increased from 9 ± 0 to 10 ± 1 (P = 0.005). Finally, training with the UroMentor(TM) resulted in a GRS increase from (1.3 ± 0.2) points to (3.9 ± 0.2) points (P < 0.001).

CONCLUSIONthe VRS UroMentor(TM) can improve urologists' ability to perform flexible cystoscopy and could be used as an effective training tool for trainees.

Clinical Competence ; Computer Simulation ; Cystoscopy ; education ; Humans ; Urology ; education

Objective To study the bone biomechanics of the rabbit osteoporosis models induced by dexamethasone sodium phosphate injection (DX) using a combined treatment modality of 99Tc-MDP and GuKangLing.Methods Rabbits were intramuscularly injected with DX (2 mg/kg) twice a week for 6 weeks.The animal osteoporosis model group (Group C) and normal group (Group A) were compared to confirm the model was available.Another control group (Group B),the osteoporosis control group (Group D) were set for the comparison at the end of the experiment.The 99Tc-MDP therapy group (Group E),GuKangLing therapy group (Group F) and 99Tc-MDP plus GuKangLing therapy group (Group G) were included in the study.The treatment lasted for 16 weeks.The bone biomechanics,cytopathology bone histomorphology,bone mineral density (BMD),X-ray,CT,bone scintigraphy and serum bone alkaline phosphatase (BALP)and P (bone gla protein) were chosen as the markers or methods to evaluate the treatment results (excellent,effective and invalid).The analysis of variance (ANOVA) and t-test were used for group comparison analysis.Results Cytopathology result indicated that there was no bone trabecula destruction in Group A.However,there was distinct bone destruction in Group C.The bone biomechanics (left femur head (265.914 ±52.773) N,L4(369.671 ±94.919) N),BMD(left femur (0.238 ±0.016) g/cm2,L4(0.236 ±0.016) g/cm2)and bone histomorphology ( (66.230 ± 10.848) ％ ) in Group C reduced clearly as compared with Group A ((405.343±55.410) N,(750.870±53.718) N,(0.294±0.017) g/cm2,(0.302±0.023) g/cm2,( 131.500 ± 21.846) ％ ) ( t ≥4.550,all P ＜ 0.01 ).Radionuclide bone scan also showed that the uptake of tracers was higher by the main arthrosis in Group C than that in Group A.Vertebra was not clearly visualized on bone scan image.There were significant differences between Group A and Group C in serum BALP and P ((45.000±7.303) vs (12.485 ±1.512) U/L,(0.168±0.018) vs (0.115 ±0.017) μg/L,t =4.126,5.476,both P ＜ 0.01 ),which indicated that the animal osteoporosis model was available.The pathological results showed an improved recovery of bone structure and trabecular in Groups E and G,but a worse recovery in Group F.Biomechanics result in Groups E and G (left femur head (386.457 ±77.077) N and (432.771 ± 17.525) N,L4(649.550 ± 126.859) N and (655.443 ±76.555) N) improved apparently,which were similar to Group B.The radiotracer uptake in Group F was lower than that in group D.The bone biomechanics,bone histomorphology,BMD,serum BALP and P after the treatment showed significant differences in Groups E,F and G (F:8.556 - 31.608,all P＜0.01 ),and the bone biomechanics result in Group G was a little better than that in Group E (t =2.625,P ＜ 0.05 ).The results of Group G and E were considered as excellent,and Group F was considered as effective.Conclusions The treatment of 99Tc-MDP combined with GuKangLing could improve the bone biomechanics of rabbit osteoporosis models and may be a potential method to increase the bone strength for resisting external force.