P450-mediated dehydrotyrosine formation during WS9326 biosynthesis proceeds via dehydrogenation of a specific acylated dipeptide substrate

WS9326A is a peptide antibiotic containing a highly unusual N-methyl-E-2-3-dehydrotyrosine (NMet-Dht) residue that is incorporated during peptide assembly on a non-ribosomal peptide synthetase (NRPS). The cytochrome P450 encoded by sas16 (P450Sas) has been shown to be essential for the formation of the alkene moiety in NMet-Dht, but the timing and mechanism of the P450Sas-mediated α,β-dehydrogenation of Dht remained unclear. Here, we show that the substrate of P450Sas is the NRPS-associated peptidyl carrier protein (PCP)-bound dipeptide intermediate (Z)-2-pent-1'-enyl-cinnamoyl-Thr-N-Me-Tyr. We demonstrate that P450Sas-mediated incorporation of the double bond follows N-methylation of the Tyr by the N-methyl transferase domain found within the NRPS, and further that P450Sas appears to be specific for substrates containing the (Z)-2-pent-1'-enyl-cinnamoyl group. A crystal structure of P450Sas reveals differences between P450Sas and other P450s involved in the modification of NRPS-associated substrates, including the substitution of the canonical active site alcohol residue with a phenylalanine (F250), which in turn is critical to P450Sas activity and WS9326A biosynthesis. Together, our results suggest that P450Sas catalyses the direct dehydrogenation of the NRPS-bound dipeptide substrate, thus expanding the repertoire of P450 enzymes that can be used to produce biologically active peptides.

Improved development of somatic cell cloned bovine embryos by a mammary gland epithelia cells in vitro model

Previous studies have established a bovine mammary gland epithelia cells in vitro model by the adenovirus-mediated telomerase (hTERT-bMGEs). The present study was conducted to confirm whether hTERT-bMGEs were effective target cells to improve the efficiency of transgenic expression and somatic cell nuclear transfer (SCNT). To accomplish this, a mammary-specific vector encoding human lysozyme and green fluorescent protein was used to verify the transgenic efficiency of hTERT-bMGEs, and untreated bovine mammary gland epithelial cells (bMGEs) were used as a control group. The results showed that the hTERT-bMGEs group had much higher transgenic efficiency and protein expression than the bMGEs group. Furthermore, the nontransgenic and transgenic hTERT-bMGEs were used as donor cells to evaluate the efficiency of SCNT. There were no significant differences in rates of cleavage or blastocysts or hatched blastocysts of cloned embryos from nontransgenic hTERT-bMGEs at passage 18 and 28 groups (82.8% vs. 81.9%, 28.6% vs. 24.8%, 58.6% vs. 55.3%, respectively) and the transgenic group (80.8%, 26.5% and 53.4%); however, they were significantly higher than the bMGEs group (71.2%, 12.8% and 14.8%), (p < 0.05). We confirmed that hTERT-bMGEs could serve as effective target cells for improving development of somatic cell cloned cattle embryos.

Association of miR-199a expression with clinicopathologic characteristics and prognosis of renal cell carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the association of miR-199a expression with the clinicopathologic characteristics and the survival of patients with renal cell carcinoma (RCC).</p><p><b>METHODS</b>Real-time quantitative RT-PCR was used to detect the expression of miR-199a in the tumor tissues and paired adjacent normal tissues from 67 patients with RCC. The correlations of miR-199a expression with the clinicopathologic characteristics and survival of RCC patients were analyzed.</p><p><b>RESULTS</b>Positive miR-199a expression was detected in both the RCC and adjacent normal tissues, but the tumor tissues showed a significantly lower expression level by a mean of 13.7 folds (P=0.003). The expression of miR-199a was negatively correlated with tumor recurrence in RCC patients in T stage (P<0.05), but showed no significant correlations with the patients age, histological type of the tumor, lymph node metastasis, distal metastasis, or Fuhrman grade (P>0.05). The patients with lowered miR-199a expression in the tumor tissue had a significantly shorter mean survival time than those without miR-199a down-regulation (P=0.017 by Log-rank test).</p><p><b>CONCLUSION</b>A decreased expression of miR-199a is significantly correlated with a higher tumor stage, a greater likeliness of tumor recurrence, and a poorer prognosis in RCC patients. miR-199a can serve as a promising prognostic factor of renal cell carcinoma.</p>

Acupuncture and Rehabilitation Training for Spastic Cerebral Palsy / 中国康复理论与实践

@#Objective To observe clinical effect of acupuncture combined with rehabilitation training on spastic cerebral palsy. Methods 160 patients with cerebral palsy were divided into acupuncture group(n=80) and control group(n=80). The acupuncture group received acupuncture, including the acupoints such as Baihui, Sishengcong, Yamen, Fengchi, etc., and combined with function training. The control group received Bobath appoach as rehabilitation training. Two groups were treated for 3 months. Results The acupuncture group(n=77) showed powerfully for 26 cases, effective for 46 cases, and invalid for 5 cases, and the efficient rate was 93.5%. The control group (n=78) showed powerfully for 20 cases, effective for 45 cases, invalid for 13 cases, and the efficient rate was 83.3% (P<0.05). Conclusion Acupuncture combined with rehabilitation training can facillitate to reduce muscle tone of spastic cerebral palsy, and improve effect of rehabilitation.

S3 neuromodulation using Chinese electro-acupuncture on BL-33 to treat neurogenic detrusor overactivity in patients with spinal cord injury / 中华泌尿外科杂志

Objective To observe the effects of electrical acupuncture for the treatment of neurogenic detrusor overactivity (DO) caused by spinal cord injury (SCI).Methods Twenty SCI patients with DO were included in this study. There were 18 males and 2 females;age ranged from 17 to 58 years. Patients were given electro-acupuncture treatment at the bilateral S3 foramen at the lateral position during the vidio-urodynamic investigation when the detrusor of the patients began to contract.The different intensities of electrical stimulation were used when the DO appeared during cystometry and the most effective intensity to inhibit DO was determined. Then, the bladder was emptied and the stimulation with selected intensity was used at the beginning of cystometry. The changes of parameters in the urodynamics, the urinary incontinence times per day and the pad used per day were recorded. The acute effects were observed. Ten days were set as one course and after 3 courses and 9 courses the patients underwent urodynamic test again. The long-term effects were observed.Results After treatment, the urinary incontinence times per day and the pad used per day decreased obviously (P<0. 05). The bladder capacity increased significantly and the maximum intravesical pressure decreased significantly. There were 3 patients having the long terms treatment. Conclusions The Chinese electro-acupuncture at S3 foramen in the SCI patients with DO is demonstrated effective. After the treatment bladder capacity could increase and the times of the urinary incontinence per week decrease.

Effect Of Electroacupuncture on Migraine / 中国康复理论与实践

@# Objective To observe the effect of electroacupuncture on migraine. Methods 60 patients were divided into electroacupuncture group (30 cases) who received electroacupuncture, and control group (30 cases) who received local nerve block. Results In the electroacupuncture group, 22 (73.33%) cases were cured, 6 (20.00%) cases were obviously improved, 2 (6.67%) cases were improved. While in control group, 17 (56.67%) cases were cured, 5 (16.67%) cases were obviously improved, 8 (26.67%) cases were improved. There was significant difference between these two groups (P<0.05). The time of treatment was less in the electroacupuncture group than in the control (P<0.05). Conclusion Electroacupuncture is effective on migraine.

Clinical observation on nanometer acupoint mounting method for alleviation of myospasm complicated by spinal injury / 中国针灸

<p><b>OBJECTIVE</b>To observe clinical effect of nanometer acupoint mounting method for alleviation of myospasm complicated by spinal injury.</p><p><b>METHODS</b>Sixty cases were randomly divided into an observation group and a control group, 30 cases in each group. The observation group were treated by nanometer mounting at 4 Jiaji (EX-B 2) points each on both sides of the affected spine and Shenshu (BL 23), Shangliao (BL 31), Ciliao (BL 32), Yang-lingquan (GB 34), Xuanzhong (GB 39); and the control group by mounting zinc oxide sticking tablets at the above acupoints. The mounting was replaced once each two days, 7 times constituting one course. One week and one month after the end of 3 courses, their results were recorded, respectively.</p><p><b>RESULTS</b>Before treatment, there was no significant difference between the two groups in grades of the myospasm degree (P > 0.05). One week after the end of treatment, 15 cases were grade I of myospasm, 9 cases were grade II, 5 cases were grade III and 1 case was grade IV in the observation group, and 1 cases grade I, 7 cases grade II, 14 cases grade III, 8 cases grade IV in the control group. Ridit analysis on the data indicated that there were significant differences before and after treatment in the myospasm degree (P < 0.01) and between the two groups after treatment (P < 0.01). One month after the end of treatment, the results were similar to those one week after the end of treatment.</p><p><b>CONCLUSION</b>Nanometer acupoint mounting method is a new one for alleviation of myospasm complicated by spinal injury, with convenience, safety and no side effect.</p>

Effect of Traditional Chinese Drug Combined with Training of Musculus Quadriceps Fexoris on Knee Osteoarthritis / 中国康复理论与实践

@#Objective To observe the effect of traditional Chinese drug combined with training of musculus quadriceps fexoris on knee osteoarthritis(KOA).Methods70 out-patient clinic KOA patients were divided randomly into the Chinese drug group and control group with 35 cases in each group.The Chinese drug group was treated with Shentongzhuyu medicinal broth PO bid;the control group was treated with Sulindac 0.2 g PO bid.Two groups were combined with the training of musculus quadriceps fexoris,having 10 times per course and 3 courses in total with a 3 days interval between two courses.The therapeutic effect was evaluated with footplate pressure gait analysis and modified JOA marks.ResultsAfter treatment,the effect of the Chinese drug group was superior to the control group(P<0.05),especially 12 weeks post treatment.There maximum weight loading,time integral and weight loading intergral of affected limb of the Chinese drug group significantly improved after treatment(P<0.01),but for the control group,only weight loading intergral improved(P<0.05).ConclusionTraditional Chinese drug combined with training of musculus quadriceps fexoris has better curative effect on the pain and functional disturbance of KOA.

Effect of Traditional Chinese Drug Combined with Training of Musculus Quadriceps Fexoris on Knee Osteoarthritis / 中国康复理论与实践

@#Objective To observe the effect of traditional Chinese drug combined with training of musculus quadriceps fexoris on knee osteoarthritis(KOA).Methods70 out-patient clinic KOA patients were divided randomly into the Chinese drug group and control group with 35 cases in each group.The Chinese drug group was treated with Shentongzhuyu medicinal broth PO bid;the control group was treated with Sulindac 0.2 g PO bid.Two groups were combined with the training of musculus quadriceps fexoris,having 10 times per course and 3 courses in total with a 3 days interval between two courses.The therapeutic effect was evaluated with footplate pressure gait analysis and modified JOA marks.ResultsAfter treatment,the effect of the Chinese drug group was superior to the control group(P<0.05),especially 12 weeks post treatment.There maximum weight loading,time integral and weight loading intergral of affected limb of the Chinese drug group significantly improved after treatment(P<0.01),but for the control group,only weight loading intergral improved(P<0.05).ConclusionTraditional Chinese drug combined with training of musculus quadriceps fexoris has better curative effect on the pain and functional disturbance of KOA.