Atropa belladonna is a medicinal plant and main commercial source of tropane alkaloids (TAs) including scopolamine and hyoscyamine, which are anticholine drugs widely used clinically. Based on the high throughput transcriptome sequencing results, the digital expression patterns of UniGenes representing 9 structural genes (ODC, ADC, AIH, CPA, SPDS, PMT, CYP80F1, H6H, TRII) involved in TAs biosynthesis were constructed, and simultaneously expression analysis of 4 released genes in NCBI (PMT, CYP80F1, H6H, TRII) for verification was performed using qPCR, as well as the TAs contents detection in 8 different tissues. Digital expression patterns results suggested that the 4 genes including ODC, ADC, AIH and CPA involved in the upstream pathway of TAs, and the 2 branch pathway genes including SPDS and TRII were found to be expressed in all the detected tissues with high expression level in secondary root. While the 3 TAs-pathway-specific genes including PMT, CYP80F1, H6H were only expressed in secondary roots and primary roots, mainly in secondary roots. The qPCR detection results of PMT, CYP80F1 and H6H were consistent with the digital expression patterns, but their expression levels in primary root were too low to be detected. The highest content of hyoscyamine was found in tender stems (3.364 mg x g(-1)), followed by tender leaves (1.526 mg x g(-1)), roots (1.598 mg x g(-1)), young fruits (1.271 mg x g(-1)) and fruit sepals (1.413 mg x g(-1)). The highest content of scopolamine was detected in fruit sepals (1.003 mg x g(-1)), then followed by tender stems (0.600 mg x g(-1)) and tender leaves (0.601 mg x g(-1)). Both old stems and old leaves had the lowest content of hyoscyamine and scopolamine. The gene expression profile and TAs accumulation indicated that TAs in Atropa belladonna were mainly biosynthesized in secondary root, and then transported and deposited in tender aerial parts. Screening Atropa belladonna secondary root transcriptome database will facilitate unveiling the unknown enzymatic reactions and the mechanisms of transcriptional control.
Alkaloids ; biosynthesis ; genetics ; metabolism ; Atropa belladonna ; genetics ; metabolism ; Gene Expression Regulation, Plant ; genetics ; Hyoscyamine ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism ; Scopolamine Hydrobromide ; metabolism ; Tropanes ; metabolism

OBJECTIVETo study the effect of exogenous Ca2+ on photosynthetic parameters of Pinellia ternate and accumulations of active components under high temperature stress.

METHODThe pigment contents of P. ternata leaves, photosynthesis parameters and chlorophyll fluorescence parameters of P. ternata leaves, the contents of guanosine, adenosine and polysaccharide in P. ternata tubers were measured based on different concentrations of exogenous Ca2+ in heat stress when the plant height of P. ternata was around 10 cm.

RESULTThe contents of total chlorophyll and ratio of chlorophyll a/b were relatively higher by spaying Ca2+. Compared with the control, spaying 6 mmol x L(-1) Ca2+ significantly enhanced the net photosynthetic rate (Pn), transpiration (Tr) and stomatal limitation (L8), but reduced intercellular CO2 concentration (C) in P. ternata leaves. With the increase of Ca2+ concentration, maximal PS II efficiency (Fv/Fm), actual photosynthetic efficiency (Yield) and photochemical quenching coefficient (qP) initially increased and then decreased, however, minimal fluorescence (Fo) and non-photochemical quenching coefficient (NPQ) went down first and then went up. The contents of guanosine and polysaccharide and dry weight of P. ternata tubers showed a tendency of increase after decrease, and the content of adenosine increased with the increase of Ca2+ concentration. The content of guanosine and polysaccharide in P. ternata tubers and its dry weight reached maximum when spaying 6 mmol x L(-1) Ca2+.

CONCLUSIONWith the treatment of calcium ion, the inhibition of photosynthesis and the damage of PS II system were relieved in heat stress, which increased the production of P. ternata tubers.

Breeding ; Calcium ; pharmacology ; Chlorophyll ; metabolism ; Dose-Response Relationship, Drug ; Heat-Shock Response ; drug effects ; Organ Size ; drug effects ; Photosynthesis ; drug effects ; Pinellia ; drug effects ; growth & development ; metabolism ; physiology ; Plant Leaves ; drug effects ; growth & development ; metabolism

OBJECTIVETo evaluate and compare standard sperm parameters and sperm DNA fragmentation in seminal ejaculates from men whose partners had a history of recurrent pregnancy loss (RPL) and a control group of men who had recently established their fertility.

METHODSSemen samples from 85 patients with a history of RPL and 20 men with proven fertility were analyzed according to World Health Organization guidelines. Sperm DNA fragmentation was detected by sperm chromatin dispersion test (SCD).

RESULTSA significant difference (P< 0.05) was observed in sperm motility but not other parameters between the two groups. The mean number of sperm cells with fragmented DNA, represented as DNA fragmentation index, was significantly increased in the RPL group [(34.99± 14.62)%] compared with controls [(10.82± 4.80)%].

CONCLUSIONThis study has indicated that sperm from men with a history of RPL have a higher incidence of DNA damage and poor motility compared with fertile males.

Abortion, Habitual ; etiology ; genetics ; Adult ; DNA Damage ; DNA Fragmentation ; Female ; Humans ; Male ; Pregnancy ; Sperm Motility

OBJECTIVETo investigate the feasibility and efficacy of transanal specimen extraction in laparoscopic colorectal resection.

METHODSFrom January 2009 to December 2010, transanal specimen extraction technique was performed in 12 patients undergoing laparoscopic colorectal resection. The clinical data were retrospectively analyzed.

RESULTSThere were 8 males and 4 females. The age ranged from 41 to 67 (mean, 58) years old. The mean operative time was (240±45) min, mean blood loss was (70±40) ml. There was no anastomotic leakage or stenosis. One patient with constipation suffered from intractable right lower abdominal pain and was managed with antispasmodic agents. The frequency of bowel movements was 3-7/d in the 5 patients with constipation and 1-2/d in patients with colorectal carcinoma at six months postoperatively. No local recurrence and metastasis in patients with colorectal cancer after follow-up of 1-20 months.

CONCLUSIONTransanal specimen extraction in laparoscopic colorectal resection is feasible, safe, and reliable.

Adult ; Aged ; Anal Canal ; surgery ; Colectomy ; methods ; Colorectal Neoplasms ; surgery ; Female ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Nerve injury produces a long lasting neuropathic pain, manifested as allodynia, a decrease in pain threshold and hyperalgesia, an increase in response to noxious stimuli. The mechanism underlying the lasting abnormal pain is not well understood. Our previous works have shown that electrical tetanic stimulation of the sciatic nerve induces long-term potentiation (LTP) of C-fiber evoked field potentials in the spinal dorsal horn, which is considered as a synaptic model of pathological pain. In the present study we tested if nerve injury, which is proved to produce neuropathic pain, induced the spinal LTP in intact rats. C-fiber evoked field potentials in spinal dorsal horn produced by electrical stimulation (10-20 V, 0.5 ms, 1/min) of the sciatic nerve were recorded. For induction of LTP of C-fiber evoked field potentials, three types of noxious stimuli were applied. (1) Electrical tetanic stimulation (40 V, 0.5 ms pulses at 100 Hz for 1 s repeated four times at 10 s intervals). (2) Transection of the sciatic nerve at 4-5 mm distal to the stimulation electrode. (3) Crushing the sciatic nerve with a forceps four times at 4-5 mm distal to stimulation electrode (from distal to proximal with 1 mm spacing at 10 s intervals), which simulated electrical tetanic stimulation. Acute nerve injury was made by either transection of the sciatic nerve at the distal to the stimulating electrode or crushing the sciatic nerve. We found that nerve injury by cutting or crushing the sciatic nerve produced LTP of C-fiber evoked field potentials lasting until the end of the experiments (3-9 h), and that pretreatment of the sciatic nerve with lidocaine 10 min prior to the nerve transectoin completely blocked LTP induced by nerve transection. The nerve transection-induced LTP was blocked by NMDA receptor antagonist AP5. LTP produced by nerve transection could not be further potentiated by electrical tetanic stimulation, while LTP induced by single electrical tetanic stimulation could be further potentiated by transection of the sciatic nerve. However, when LTP was saturated by several times of electrical tetanic stimulation, nerve transection did not affect the spinal LTP. We conclude that acute nerve injury induces LTP of C-fiber evoked field potentials in intact animals and that nerve transection is more powerful than electrical tetanic stimulation for induction of the spinal LTP. The results further support the notion that LTP of C-fiber evoked field potentials may underlie neuropathic pain.
Animals ; Evoked Potentials ; physiology ; Long-Term Potentiation ; physiology ; Male ; Nerve Fibers, Unmyelinated ; physiology ; Neural Pathways ; drug effects ; physiology ; Nociceptors ; physiology ; Posterior Horn Cells ; enzymology ; physiology ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; physiology ; Spinal Cord ; physiology

Humans ; Lipomatosis, Multiple Symmetrical ; pathology ; surgery ; Male ; Middle Aged

AIMTo prepare the breviscapine liposomes and study the pharmacokinetics of breviscapine liposomes in Beagle dogs.

METHODSThe cross-over design (two periods) was employed. Six Beagle dogs were administrated a single intravenous dosage of 28 mg of breviscapine liposomes and reference preparation, respectively, scutellarin in plasma of 6 dogs at different sampling time was determined by RP-HPLC. The pharmacokinetic parameters were calculated by 3P97 program and compared by statistic analysis.

RESULTSThe mean concentration-time curves of breviscapine liposomes and reference preparation were both fitted to two-compartment model with the main pharmacokinetic parameters as follows: T 1/2 alpha were (4.4 +/- 0.7) min and (1.8 +/- 1.3) min respectively; T 1/2 beta were (55 +/- 27) min and (28 +/- 23) min respectively; V(c) were (1 580 +/- 265) mL and (2 460 +/- 2 200) mL respectively; CL(s) were (88 +/- 10) mL x min(-1) and (324 +/- 69) mL x min(-1) respectively; and AUC(0-720) were (363 +/- 42) microg x min x mL(-1) and (102 +/- 19) microg x min x mL(-1) respectively. The T 1/2 alpha, CL(s) and AUC(0-720) of breviscapine liposomes all had significant difference from those of reference preparation, after the data were examined by a one-way analysis of variance (ANOVA).

CONCLUSIONCompared with the reference preparation, breviscapine liposomes had a much more higher concentration in plasma and contained characteristic of sustained-release, which ameliorated the pharmacokinetic properties of scutellarin.

Animals ; Apigenin ; blood ; Area Under Curve ; Brain ; metabolism ; Cross-Over Studies ; Delayed-Action Preparations ; Dogs ; Drug Compounding ; Drug Stability ; Erigeron ; chemistry ; Female ; Flavonoids ; administration & dosage ; isolation & purification ; pharmacokinetics ; Glucuronates ; blood ; Injections, Intravenous ; Liposomes ; Male ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the procedure and the value of G-banding, fluorescence in sit hybridization (FISH) and comparative genomic hybridization (CGH) techniques in prenatal diagnosis.

METHODSKaryotype analyses with three diagnostic procedures, G-banding, G-banding and FISH, G-banding, FISH and CGH, were performed in the amniotic fluid samples taken from 102 fetuses at gestational ages 16-24 weeks. And the significance was valued in prenatal diagnosis.

RESULTSIn the first procedure of karyotype analysis, 98 cases were diagnosed, 2 cases were not conformed while 2 cases were failed in all 102 cases. In the second procedure, 2 cases were determined, 1 case was not conformed and 1 case was still failed. In the third step, 2 cases were diagnosed. The diagnostic rate of the karyotype reached to 100% (102/102 cases) using all the three procedures. In total, seven cases with chromosomal abnormality were diagnosed. Four cases, 1 case and 2 cases were identified in the first step (4/7, 57.1%), the second (1/7, 14.3%) and the third (2/7, 28.5%), respectively.

CONCLUSIONIt can help improve the diagnostic rate of chromosomal aberrations and standardize diagnostic procedure to perform the three detecting steps in prenatal diagnosis.

Chromosome Aberrations ; statistics & numerical data ; Chromosome Banding ; methods ; Chromosome Disorders ; diagnosis ; genetics ; Chromosomes, Human, Pair 18 ; Comparative Genomic Hybridization ; methods ; Female ; Fetus ; Gestational Age ; Humans ; In Situ Hybridization, Fluorescence ; Intellectual Disability ; genetics ; Karyotyping ; methods ; Male ; Nucleic Acid Hybridization ; methods ; Pregnancy ; Prenatal Diagnosis ; statistics & numerical data ; Risk Factors ; Ultrasonography, Prenatal ; methods

Adult ; Female ; Humans ; Laparoscopy ; methods ; Male ; Muscular Diseases ; surgery ; Neck Muscles ; surgery ; Syndrome

OBJECTIVE: To explore the value of ultrasonographic evaluation in fetal deformity in prenatal diagnosis by a systematic continuous sequence approach (SCSA). METHODS: Successive prenatal ultrasonographic evaluation was performed to monitor the whole anatomic structure,form, posture and movement of 16,685 fetuses during gestation aging 14 approximately 40(+3) weeks. RESULTS: Satisfactory ultrasonic images were obtained in 16,627 fetuses using the SCSA (99.65%). Of them, 514 abnormal fetuses were confirmed after subsequent labor or induced labor and 498 abnormal fetuses were correctly diagnosed using SCSA during prenatal stage (96.89%). Whereas 16 fetuses missed recognition (3.11%). Its sensitivity, specificity, positive and negative predictive value of diagnosis on fetal deformity were 96.98%, 99.96%, 98.66%, and 99.90 %, respectively. CONCLUSION SCSA in prenatal ultrasonographic evaluation of the fetal structure and malformation is reliable and accurate.
Adult ; Congenital Abnormalities ; diagnostic imaging ; Female ; Humans ; Pregnancy ; Sensitivity and Specificity ; Ultrasonography, Prenatal ; methods