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italic>Gentiana crassicaulis Duthie ex Burk. is one of the plant sources of Gentianae Macrophyllae Radix (QinJiao). Gentiana tibetica King ex Hook. f. and Gentiana robusta King ex Hook. f. are relative species of G. crassicaulis. Due to the large intraspecific morphological variation, G. crassicaulis showed high morphological similarity with G. tibetica and G. robusta. And the distribution area of the three species overlaps to some extent, which makes it difficult to identify them. On the basis of morphological identification, the method of molecular identification of the three species was constructed in this study based on chloroplast genomes. The chloroplast genome of Gentiana tibetica is 148 765bp long, with LSC, SSC and IR 81 163 bp, 17 070 bp and 25 266 bp, respectively. The structure of the three is consistent. The chloroplast genome sequences of G. tibetica and G. crassicaulis are highly similar, and the number of variable sites is 9 (149 267 bp in total). Diagnostic SNP that could effectively identify the three species was screened and verified, and a dual-peak SNP detection method was established for the effective identification of each species and mixed samples. Our study provides basic data for the molecular identification of G. crassicaulis and its related species, and the arrangement of related Tibetan medicine.
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OBJECTIVE@#To compare the effect of acupoint injection and intramuscular injection with mouse nerve growth factor (mNGF) on gross motor function development of children with cerebral palsy (CP), and explore the treatment mechanism.@*METHODS@#A total of 63 children with CP were randomly divided into an observation group (32 cases, 4 cases dropped off ) and a control group (31 cases, 3 cases dropped off). Based on the routine rehabilitation therapy, the control group received intramuscular injection of mNGF(18 µg/2 mL), and the observation group received acupoint injection of mNGF at Xinshu (BL 15), Ganshu (BL 18), Pishu (BL 20), Shenshu (BL 23), Sanjiaoshu (BL 22), Shenting (GV 24), Baihui (GV 20), Fengfu (GV 16), Dazhui (GV 14), etc. Of them, 5-6 acupoints alternately were selected each time, and each acupoint was given 0.3-0.5 mL, totally 18 µg/2 mL. Both treatment were carried out once every other day for six months. Before and after treatment, the children's development of brain function was assessed using gross motor function classification system (GMFCS). Before treatment (T), after 2 (T), 4 (T) and 6 (T) months of treatment, the motor function was evaluated by gross motor function measure (GMFM-88). The systolic peak velocity (Vs), mean velocity (Vm) and vascular resistance index (RI) of anterior cerebral artery (ACA) and middle cerebral artery (MCA) were measured, and the level of N-acetyl aspartate acid (NAA), choline (Cho), lactate (Lac) and creatine (Cr) from the basal ganglia, thalamus and periventricular white mater were detected by magnetic resonance spectroscopy (MRS) technology with MAGNETOM Skyra3.0T magnetic resonance imaging system before and after treatment.@*RESULTS@#Compared with before treatment, the GMFCS classification of the observation group after treatment was significantly improved (0.05), however, the observation group had a 3.142 times of feasibility for good gross motor function development by more than level 1 compared to the control group (<0.05). After 2, 4, and 6 months of treatment, the GMFM-88 scores of the two groups showed an upward trend (<0.01), and the increase of the observation group was greater than that of the control group (<0.05). Compared with before treatment, in the ACA and MCA, the Vs and Vm increased, RI decreased in both groups after treatment (<0.01), and in the brain, NAA/Cr increased, Cho/Cr and Lac/Cr decreased (<0.01), and after treatment, the Vs, Vm of ACA and MCA and NAA/Cr of brain in the observation group were higher than those in the control group (<0.05), and the RI of ACA and MCA and Cho/Cr and Lac/Cr of brain in the observation group were lower than those in the control group (<0.05).@*CONCLUSION@#The mNGF acupoint injection has a better effect on the gross motor function in the children with cerebral palsy compared with the intramuscular injection, and the mechanism may be associated with exhibiting the double effects of acupoint effect and the targeting therapy of drug, which can effectively improve the cerebral hemodynamics and the metabolism of cerebral nervous substances.
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As a common Tibetan herb, Bawo Sebo was mainly used in the treatment of rheumatoid arthritis and urarthritis in Traditional Tibetan medicine. Based on our ethnobotanical survey, the origin of the herb was determined as Swertia verticillifolia T. N. Ho et S. W. Liu (Gentianaceae), endemic to the region of the Qinghai-Tibet Plateau. The diagnostic characters:perennial; stem leaves in whorls; corolla campanulate, yellow-green, 4-lobed; nectary 1 per corolla lobe, naked. Also, its complete chloroplast (cp) genome was sequenced. It is 151 682 bp in length, including a large single copy (LSC) region of 82 623 bp, a small single copy (SSC) region of 18 335 bp and a pair of inverted repeats (IRs) of 25 362 bp. It contains 129 unique genes, including 84 protein-coding genes, 37 tRNAs and 8 rRNAs. This study provides information for understanding the diversity of Swertia cp genomes, and the alpine species identification, conservation and molecular phylogenetic researches of Swertia and Gentianaceae.
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Objective To investigate the effect of hypobaric hypoxia and cold exposure on brown adipose tissue in mice. Methods Twenty-four 6-week old SPF C57BL/6 male mice were randomly divided into 4 groups with 6 mice in each group: normal atmospheric pressure and temperature group ( 18~22℃, 20~60 m ) ( NTNP ) , low atmospheric pressure and normal temperature group ( 18~22℃, altitude of 5000 m ) ( NTLP ) , normal atmospheric pressure and cold exposure group(0~6℃, altitude of 20 ~60 m)(LTNP), low atmospheric pressure and cold exposure group(0 ~6℃, altitude of 5,000 m)(LTLP). The experimental period was 4 weeks. The body weight was measured at the beginning and end of the experiment. By the end of the four-week trial, the back and inguinal fat were dissected and observed by histology using HE staining. The expression of UCP-1 as the marker of brown adipose tissue in the back fat was detected by qPCR and western blot. Results The body weight gain of NTNP group was higher ( P< 0. 05 ) than the other three groups. Meanwhile, the color of the back and groin fat tissue of mice of LTNP and LTLP groups were darker, the blood supply in mice of these two groups was richer than the NTLP group. The volume of adipose tissue of NTNP group was higher than others. The histology showed that the back adipose cells of the mice were smaller and darker and full of multilocular lipid droplets, exhibiting a typical morphology of brown fat cells. Compared with the NTNP and NTLP groups, the mRNA and protein levels of UCP-1 were higher under cold exposure, while low atmospheric pressure had a tendency to reduce the mRNA expression of UCP-1. Conclusions The formation of brown fat is affected by the imitated conditions of low atmospheric pressure and cold exposure, and is more closely related to the decresed temperature.
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The circular double-stranded cpDNA sequence consists of two inverted repeat (IR) regions, a large single-copy region (LSC) and a small single-copy region (SSC). The function of some genes within the genome is clear. However, it is unknown for such chloroplast genes as hypothetical chloroplast reading frames (ycfs) and so on. As one of ycf s, ycf15 gene is located within IR regions with length of 250-550 bp, conserved among plants and also, its function is often unclear. The gene has attracted an intense attention during the past decade. This review presents an overview of ycf15 gene including its basic type, structural features, and the application to phylogenetic analyses. As a potential DNA marker for species identification, future implications of the gene are also introduced.