AIM: To establish the determination for camphor, menthol, synthetic borneol, and methyl salicylate in four kinds of TCM adhesive plasters by GC. METHODS: The internal standard method was employed. Naphthalene, the internal standard, was added to the sample before treatment. The sample solution was prepared by distillation with essential oil collector. The GC system consisted of glass column, 10% PEG 20M as the stationary phase, nitrogen as the carrier gas, column temperature at 130℃, and FID as the detector. RESULTS: Camphor, menthol, synthetic borneol(borneol and isoborneol), and methyl salicylate in 6 lots of samples and naphthalene have been separated well under the chromatographic condition. In the methodology study on Fengshi Zhitong Adhesive Plasters, the average recovery of camphor, menthol, synthetic borneol, and methyl salicylate were 95.54%( RSD =1.6%), 96.90%( RSD =2.2%), 97.30%( RSD =1.4%), and 96.48%( RSD =2.8%), respectively. CONCLUSION: The method is sensitive, accurate and reproducible, and can be used to control the quality of the four kinds of preparations.

Objective To identify a myeloid differentiation factor 88(MyD88)in Oncomelania hupensis,and characterize the role of MyD88 against Schistosoma japonicum infection. Methods The complete cDNA of MyD88 in O. hupensis was ob-tained by using rapid amplification of cDNA ends(RACE),and homologues sequences and conserved domains were aligned and the structure of MyD88 was predicted either. A phylogenetic tree of MyD88 was further constructed with other species. In ad-dition,the mRNA expression level of O. hupensis MyD88 before and after S. japonicum infection was investigated by real-time quantitative PCR(RT-qPCR). Results The cDNA of O. hupensis MyD88 consisted of 1406 bp open reading frame(ORF),en-coding 468 amino acid residues,which contained death domain and Toll/interlrukin-1 receptor(TIR)domain,the typical fea-tures of MyD88 family proteins. The predicted amino acid sequence of O. hupensis MyD88 shared 38%-52%identity with other mollusc. O. hupensis MyD88 was phylogenetically closeted to Biomphalaria glabrata MyD88. The O. hupensis MyD88 existed in all selected tissues and expressed highly in hemocyte,up-regulated after S. japonicum infection in all selected tissues except cephalopodium,especially higher in whole snail and hemocyte. Conclusion MyD88-dependent signaling pathway is present in O. hupensis and plays an important role in innate immune response against S. japonicum infection.

Objective To discuss the effect of gene transfection of rAAV2/1-NPY-EGFP on KA-induced rat seizures,EEG and the expression of hippocampal phosphorylated Tau protein.Methods Altogether 72 healthy male Wistar adult rats were randomly divided into three groups:control group,KA group and NPY group(n=24).The epileptic models were established by the injection of KA 2 μl (0.4 μg/μl) five times to the right side of the hippocampus CA3 area every three days.rAAV2/1-NPY-EGFP group,in which 10 μl of rAAV2/1-NPY-EGFP (titer 5× 1011 v.g./ml) was injected to the lateral ventricle in successful rats chronic model,while KA group was injected with an equal dose of saline.The control group was injected with an equal dose of saline both in the hippocampal CA3 area and the lateral ventricle.The seizure situation,the onset latency and EEG were observed at 2 weeks and 4 weeks after vector injection.Then the expression of phosphorylated Tau protein in hippocampus were detected with Western blotting.Results (1) Scale and latency of each seizure onset in rats of rAAV2/1-NPY-EGFP group (12.13 ± 8.06) had no significant difference at 2 weeks (P＞ 0.05) compared with KA group (12.10± 8.07).The scale of seizure in rats of rAAV2/1-NPY-EGFP group(6.06±3.78) significantly reduced at 4 weeks(P ＜0.05).Latency of seizure onset (79.06±8.83min) significantly increased at 4 weeks(P＜0.05),EEG epileptic discharge frequency and wave amplitude decreased (P＜ 0.05) at 4 weeks.The control group had no seizures.(2)Compared with the control group,the expression of phosphorylated Tau protein in KA group and NPY group significantly increased(P＜0.05) at 2 weeks and 4 weeks,and the expression of phosphorylated Tau protein in the NPY group (1.15±0.16 RQ value) at 4 weeks significantly decreased (P＜0.05) compared with the KA group(1.87± 0.23 RQ value).Conclusion rAAV2/1-NPY-EGFP gene transfection significantly reduces scale of seizure onset and prolongs latency of seizure onset in KA-induced rat model.rAAV2/1-NPY-EGFP gene transfection may play anti-epileptic and neuroprotective effects through inhibiting the expression of phosphorylated Tau protein in hippocampus of KA-induced epileptic rat model.

Objective To observe the cerebral hemodynamics in ischemic stroke patients after compound anisodine acupoint injection.Methods 40 stroke patients were measured with transcranial color Doppler before and 4 h,24 h after compound anisodine acupoint injection.Results After injection,the velocity of the cerebral artery showed some increase,the end diastolic velocity of the ophthalmic artery increased and the pulsitility index decreased(P<0.05).Conclusion The compound anisodine acupoint injection may improve the flow velocity of ophthalmic artery and cerebral artery.

Objective To investigate the molecular characterization of a Chinese pedigree with rare β thalassemia genotype.Methods Phenotypic analysis was performed using standard hematological tests to measure red blood cell parameters, including RBC,Hb,MCV,MCH,MCHC and RDW.SPIFE automatic Hb agarose gel electrophoresis instrument was used to measure hemoglobin fraction Hb A,Hb A2 and Hb F.The alleles of β thalassemia mutation were determined by RDB assay, and then cloning and sequencing were performed to define the mutation sites.Results The proband and his father had typical microcytic hypochromic anemia with low MCV and MCH(79.8, 63.1 fl and 19.9, 20.9 pg, respectively) and high level of Hb A2 (5.66% and 5.60%, respectively).The proband′s mother had normal MCV and MCH. β thalassemia mutation analysis with RDB assay showed that the proband had thalassemia minor resulting from double mutations on one globin gene.One showed codons 41/42 (-TTCT) mutation and the other was CAP mutation from positions +40 to +43 in the promoter region.These two mutations were inherited from his father.The genotype of the proband and his father was β41/42、CAP/βA ,and the genotype of his mother was βA/βA.Conclusions It′s rare that double mutations occur on single β globin gene, with one mutation on CD41/42(-TTCT) and the other mutation from positions +40 to +43 relative to the mRNA cap site in the promoter region.The findings enrich knowledge of the mutation spectrum of β thalassemia.

Objective To compare the dosimetry difference among three-dimensional conformal radiotherapy (3 DCRT),simplified intensity-modulated radiotherapy (sIMRT) and intensity-modulated radiotherapy (IMRT) in whole pelvic irradiation in postoperative rectal carcinoma,in order to optimize the protocol for clinical practice.Methods From 2006 to 2008,10 patients with stage II and ID rectal cancer after radical resection (Dixon surgery) participated in this study.3DCRT,sIMRT and IMRT were performed for each patient.The dose distribution of target volume and normal tissues,conformal index (CI) and HI were analyzed using the dose-volume histogram (DVH).Results The CI for PTV of IMRT and sIMRT was superior to that of 3DCRT.3DCRT had the best HI in PTV target area dose distribution,while IMRT was similar with sIMRT,however,there were no significant difference among them.As regarded as the protection on organs at risk,for bladder,IMRT was superior to 3DCRT and slightly better than sIMRT;for small intestine,sIMRT showed better performance than 3DCRT while IMRT was better than sIMRT but with no significant difference;for colon,no dosimetry difference was found among three plans;for caput femoris,IMRT and sIMRT were better than 3DCRT.Additionally,sIMRT was similar to 3DCRT in MU of segments,but significantly lower than IMRT.The mean values of total MU for 3DCRT,sIMRT and IMRT were 569.73 ±48.69,542.97 ±69.78,and 770.25 ±73.12,respectively.Conclusions All of 3DCRT,sIMRT and IMRT could provide target area with sufficient and accurate dose,meanwhile they could also protect organs at risk well on rectal cancer after radical resection.Compared with 3DCRT plan and IMRT plan,sIMRT plan might be the optimal plan for clinical practice.

Objective To evaluate the efficacy and safety of adding neoadjuvant chemotherapy following neoadjuvant chemoradiotherapy in patients with locally advanced rectal cancer.Methods A total of 80 patients confirmed with locally advanced rectal cancer were enrolled during January 2012 and December 2015 in Guizhou Medical University Affiliated Cancer Hospital and were randomized with the method of lottery into the experimental group and the control group.In the experimental group,40 patients received 4 cycles of FOLFOX4 after chemoradiotherapy and then had total mesorectal excision (TME).Another 4 cycles of FOLFOX4 were administered after surgery.In the control group,40 patients had TME surgery 6-8 weeks after chemoradiotherapy and received 8 cycles of FOLFOX4 as adjuvant chemotherapy.Pelvic radiotherapy dose was 50 Gy in 25 fractions,5 days per week for 5 weeks.5-Fu continuous infusion was administered throughout radiotherapy.The pCR rate,downstaging rate,R0 resection rate,local recurrence rate,distant metastasis rate,survival rate,incidence of toxicities,surgical complications and completion of treatment were observed.Results The pCR rate was 20.0％ in the experimental group and 5.0％ in the control group (x2 =4.114,P ＜ 0.05).The downstaging rate was 77.4％ in the experimental group and 55.6％ in the control group(P ＞ 0.05).No statistically significant difference was observed in R0 resection rate,3-year local recurrence rate,3-year distant metastasis rate and 3-year survival rate between the two groups (P ＞ 0.05).Patients in the experimental group had higher completion rate of 8 cycles of systemic chemotherapy (87.1％ vs.61.5％,x2 =4.985,P ＜0.05).No statistically significant difference was observed in acute toxicities and postoperative complications.Conclusions Adding systemic chemotherapy after neoadjuvant chemoradiotherapy for locally advanced rectal cancer has significantly higher pCR rate and lower toxicities and side events compared with chemoradiotherapy alone.Longer follow-up and larger scale of clinical research are needed.Trial registration Chinese clinical trial registry,ChiCTR-IPR-17010454.

ObjectiveTo explore the clinic symptom and the characteristics of video,tightly close,intracranial electroencephalogram (EEG) of patients with central region diastematia epilepsy. Methods Retrospective analysis of 9 patients with central region diastematia epilepsy admitted from June,2007 to August,2009.The characteristics of all patients' seizure symptom and EEG manifestation were analyzed using patients' medical history,video and EEG records.ResultsPatients with central region diastematia epilepsy had relatively long sezure history.The duration of seizure was commonly short,with frequent episode and no obvious intelligence impairment.The seizure was often accompanied with the hyperkinesia in the lower limbs.Scalp EEG showed discharges with low amplitude waves in the mean line area.The superhigh amplitude and regular rhythm slow sharp wave could be found in the diastematia cortex EEG.All patients had an Engel Class Ⅰ outcome after surgery.ConclusionThe seizure symptoms are characteristic in the patients with central region diastematia epilepsy,and some special manifestations can be found in different phase,wave amplitude,rhythm,lead array.

2-Keto-L-gulonic acid (2-KLG), the precurcor of L-ascorbic acid synthesis, was prepared directly from D-glucose by tandem fermentation. In the first step fermentation Erwinia sp. SCB 247 translated D-glucose to 2,5-diketo-D-gluconate (2,5-DKG), which accumenlated 180 mg 2,5-DKG per ml in the broth. In the second step fermentation Corynebacterium sp. SCB 3058 reduced 2,5-DKG to 2-KLG, which accumulated 35 mg 2-KLG per ml in the broth. This reductive fermentation was obtained under aerobic conditions by adding a hydrogen donor such as glucose.The average yield of five batches fermentation was 56.3 mol%, from D-glucose to 2-KLG in 10 L fermentor.

OBJECTIVETo determine mutations of two common potassium channel subunit genes KCNQ1, KCNH2 causing long QT syndrome (LQTS) in the Chinese.

METHODSThirty-one Chinese LQTS pedigrees were characterized for mutations in the two LQTS genes, KCNQ1 and KCNH2, by sequencing.

RESULTSTwo novel KCNQ1 mutations, S277L in the S5 domain and G306V in the channel pore, and two novel KCNH2 mutations, L413P in the transmembrane domain S1 and L559H in the transmembrane domain S5 were identified. The triggering factors for cardiac events developed in these mutation carriers included physical exercise and excitation. Mutation L413P in KCNH2 was associated with the notched T wave on ECGs. Mutation L559H in KCNH2 was associated with the typical bifid T wave on ECGs. Mutation S277L in KCNQ1 was associated with a high-amplitude T wave and G306V was associated with a low-amplitude T wave. Two likely polymorphisms, IVS11 + 18C > T in KCNQ1 and L520V in KCNH2 were also identified in two LQTS patients.

CONCLUSIONSThe mutation rates for both KCNQ1 (6.4%) and KCNH2 (6.4%) are lower in the Chinese population than those from North America or Europe.

Asian Continental Ancestry Group ; Cation Transport Proteins ; China ; DNA-Binding Proteins ; ERG1 Potassium Channel ; Ether-A-Go-Go Potassium Channels ; Female ; Humans ; KCNQ Potassium Channels ; KCNQ1 Potassium Channel ; Long QT Syndrome ; genetics ; Male ; Mutation ; Potassium Channels ; genetics ; Potassium Channels, Voltage-Gated ; Trans-Activators ; Transcriptional Regulator ERG