Evaluation of the Optimal Duration of Thumb Cupping Therapy for Frozen Shoulder by Thermal Metabolic Imaging.

Objective: To evaluate the optimal duration of thumb cupping therapy for frozen shoulder (FS) by thermal metabolic imaging (TMI) and to provide clinical evidence. Methods: From April 2022 to August 2023, 120 patients with FS who received thumb cupping therapy at our hospital were randomized into groups A, B, and C for 5, 10, and 15 minutes of cupping, respectively. The clinical efficacy, Visual Analogue Scale (VAS) score, and shoulder range of motion (ROM) of the three groups were compared, and the skin blood flow and the incidence of adverse reactions during treatment were analyzed. Finally, the temperature difference improvement efficiency (i.e., higher TMI after treatment than before treatment) was compared among the three groups. Results: Groups B and C showed higher overall clinical efficacy than group A (P < .05). After treatment, lower VAS scores were determined in groups B and C compared with group A, whereas the ROM values of groups A and C were smaller than those of group B Group C had the greatest skin blood flow and the highest incidence of adverse reactions (P < .05), while group B had the highest temperature difference improvement efficiency (P < .05). Conclusions: Through TMI, it is confirmed that thumb cupping therapy with a duration of 10 min contributes to the highest efficacy and safety for patients with FS.

First Report of Root Rot Caused by Ilyonectria robusta in the Medicinal Herb Aconitum carmichaelii in China.

Aconitum carmichaelii Debeaux is used as a traditional Chinese medicine with antiarrhythmic, antiinflammatory and other pharmacological functions. It is widely cultivated in China. According to our survey, about 60% of A. carmichaelii in Qingchuan, Sichuan, suffered from root rot, reducing yields by 30% in the past five years. Symptomatic plants exhibited stunted growth, dark brown roots, reduced root biomass, and fewer root hairs. The disease caused root rot and plant death in 50% of the infected plants. In October 2019, ten symptomatic 6-month-old plants were collected from fields in Qingchuan. Diseased pieces of the roots were surface sterilized with sodium hypochlorite solution (2%), rinsed three times in sterile water, plated on potato dextrose agar (PDA), and incubated at 25°C in the dark. Six single-spore isolates of a Cylindrocarpon-like anamorp were obtained. The colonies on PDA were 35 to 37 mm diam after seven days with regular margins. The plates were covered with felty aerial mycelium, white to buff, and the reverse side chestnut near center with a ochre to yellowish leading edge. On spezieller nährstoffarmer agar (SNA), macroconidia were 1 to 3 septate, straight or slightly curved, cylindrical, with rounded ends, and varied in size: 1-septate 15.1 to 33.5 × 3.7 to 7.3 µm (n=250), 2-septate 16.5 to 48.5 × 3.7 to 7.6 µm (n=85), and 3-septate 22.0 to 50.6 × 4.9 to 7.4 µm (n=115). Microconidia were ellipsoid to ovoid, and 0 to 1 septate; aseptate spores were 4.5 to 16.8 × 1.6 to 4.9 µm (n=200), and 1-septate spores were 7.4 to 20.0 × 2.4 to 5.1 µm (n=200). The chlamydospores were brown, thick-walled, globose to subglobose, 7.9 to 15.9 µm (n=50). The morphology of these isolates was consistent with the previous description of Ilyonectria robusta (Cabral et al. 2012). Isolate QW1901 was characterized by sequencing the ITS, TUB, H3, and tef1α loci using previously reported primer pairs: ITS1/ITS4 (White et al. 1990), T1/Bt-2b (O'Donnell and Cigelnik 1997), CYLH3F/CYLH3R (Crous et al. 2004), and EF1/EF2 (O'Donnell et al. 1998). A Blastn search of the sequences of ITS, TUB, H3, and tef1α showed that QW1901 shared 99.26, 97.89, 97.79, and 99.17 % identities, respectively, with the ex-type strain of I. robusta (CBS308.35). The ITS, TUB, H3, and tef1α sequences were deposited in GenBank under accession nos. MW534715, and MW880180 to MW880182, respectively. A phylogenetic tree was constructed from a neighbor-joining analysis on the alignment of the combined ITS, TUB, H3, and tef1α sequence. QW1901 was clustered with the ex-type strain of I. robusta. To confirm the pathogenicity of I. robusta, bare roots of healthy 6-month-old A. carmichaelii were inoculated with mycelial plugs of 7-day-old QW1901 colonies selected randomly (Lu et al. 2015). Five needle-wound lateral roots and five intact roots were inoculated as replicates with pathogen-free agar plugs as a control. Then, all plants were grown in sterile soil in a growth chamber at 20±1°C and watered regularly. Pathogenicity assays were repeated twice. After 20 days of cultivation, infected plants exhibited symptoms similar to those observed in the field. All control plants remained asymptomatic. Sequencing confirmed the re-isolation of I. robusta from the inoculated plants, satisfying Koch's hypothesis. Ilyonectria robusta has been reported to cause root rot of plants such as Codonopsis tangshen and Panax ginseng ( Lu et al. 2015; Zheng et al. 2021), and has also been reported to be isolated from Aconitum kongboense in China (Wang et al. 2015). However, this is the first report of the pathogen causing root rot of A. carmichaelii. Management measures, such as growing disease-free seedlings in sterile soil, should be used to minimize the risk of this pathogen.

Transcriptome analysis of Aconitum carmichaelii and exploration of the salsolinol biosynthetic pathway.

Aconitum carmichaelii has been used in traditional Chinese medicine for treating various diseases for several thousand years. Based on the biosynthetic pathway of some alkaloids such as C19-diterpenoid alkaloids and obvious differences in alkaloid content between leaves of two A. carmichaelii varieties has been reported, we performed leaves transcriptome analysis of two A. carmichaelii varieties. Besides we characterized the biosynthetic pathway of salsolinol. A total of 56 million raw reads (8.28 G) and 55 million clean reads (8.24 G) were obtained from two varieties (Z175 and R184) leaves transcriptome, respectively, and 176,793 unigenes were annotated. 281 and 843 unigenes are involved in the salsolinol biosynthetic pathway and the formation of C19-diterpenoid alkaloids respectively. And including 34 and 24 unigenes are the differentially expressed genes (DEGs) in the biosynthesis pathway for C19-diterpenoid alkaloids and salsolinol between Z175 and R184 respectively, which were target genes to explore differences in C19-diterpenoid alkaloid and salsolinol biosynthesis in Z175 and R184. Thus genes involved in alkaloid biosynthesis and accumulation differ between varieties leaves. The mechanisms underlying the differences and their relevance require further exploration. The results expand our knowledge of alkaloids biosynthesis in A. carmichaelii leaves, and provide a theoretical basis for analysis differences in alkaloids biosynthesis patterns in different varieties.

Immune regulation mechanism of vitamin D level and IL-17/IL-17R pathway in Crohn's disease.

Immune regulation mechanism of vitamin D level and interleukin (IL)-17/IL-17 receptor (IL-17R) pathway in Crohn's disease was studied. Of 40 clean mature healthy rats, 10 rats were used as control group based on random number table, the remaining 30 rats to establish Crohn's disease rat models. After successful modeling, 30 rats were divided into model group, low-dose group and high-dose group with random number table. On the 1st day after modeling, rats in low-dose group were given a single dose of 1,750 IU of vitamin D, and rats in high-dose group a single dose of 7,500 IU of vitamin D. Changes in the condition of rats after modeling were observed and scored. Enzyme-linked immunosorbent assay was used for detecting IL-12, IL-17 and CXCL11 levels, western blotting for detecting IL-17R level, and flow cytometry for detecting Th1 cell and Th17 cell levels in the lamina propria of colon mucosa. Disease activity index scores were significantly lower in low-dose group and high-dose group of rats than those in model group (P<0.05). Those were significantly lower in high-dose group of rats than those in low-dose group (P<0.05). IL-17 and IL-17R levels were significantly lower in high-dose group of rats than those in low-dose group (P<0.05). Th1 cell level was significantly higher in high-dose group of rats than that in low-dose group (P<0.05), but Th17 cell level was lower than that in low-dose group (P<0.05). IL-12 levels were significantly higher in model group, low-dose group and highdose group of rats than those in control group (P<0.05). CXCL11 levels were significantly lower in model group, low-dose group and high-dose group of rats than those in control group (P<0.05). Vitamin D can effectively treat Crohn's disease, which may improve the chemotaxis and differentiation of Th1 cells by inhibiting IL-17/IL-17R pathway, thereby improving immune function and reducing the severity of disease.

Comprehensive quality evaluation of the lateral root of Aconitum carmichaelii Debx. (Fuzi): Simultaneous determination of nine alkaloids and chemical fingerprinting coupled with chemometric analysis.

Amino alcohol alkaloids are the active components in the lateral root of Aconitum carmichaelii Debx. (Fuzi), and they have a variety of pharmacological activities. However, the chemical fingerprints of the ester alkaloids reported to date were mainly obtained from high-performance liquid chromatography coupled with ultraviolet detection, and it is difficult to obtain information about amino alcohol alkaloids in Fuzi from such chromatograms. In this paper, a comprehensive fingerprinting method was established using high-performance liquid chromatography coupled with an evaporative light-scattering detector for the simultaneous quantitative analysis of both the amino alcohol alkaloids and ester alkaloids. A total of 42 samples of Fuzi from four production areas were analyzed by constructing high-performance liquid chromatography fingerprints. Then, the quantitative results of the chemical fingerprints combined with chemometrics methods were employed to reveal the factors affecting the geo-authentic Fuzi and to determine characteristic components that can be used to identify these samples. The results indicated distinct differences in the alkaloid contents among samples from the four regions; the geographical origin may be the primary factor affecting the geo-authentic Fuzi, and 15 major components (including songorine, neoline, and hypaconitine, which were quantitatively determined) were found to be characteristic components for the discrimination of Fuzi samples from various regions. Neoline might be a critical component for identifying geo-authentic Fuzi. This approach is convenient, reproducible and provides a promising method for the quality evaluation of Fuzi.

[Effects of micronutrient fertilizer application on yield and quality of Aconitum carmichaeli].

OBJECTIVE: To study the effects of Fe, Zn, B and Mn fertilizer with different ratio on the yield and quality of Aconitum carmichaeli. METHOD: Field experiment with the uniform design was applied, the yield and the contents of the total alkaloids and diester-alkaloids were measured. RESULT: Fe, Zn, B and Mn fertilizer of appropriate ratio could promote the growth of vegetative organs, increase the biomass, the content of alkaloids and the yield of Aconite significantly. Fe, Zn fertilizer of highly concentrated ratio increased the proportion of first sub-roots, but inhibited the growth of other vegetative organs, the number of roots was less than that with other treatments, so it was not conducive to the formation of production. High concentration of Mn was not conducive to the growth of underground of Aconite, its number of sub-roots was fewer, but the number of third sub-roots was more than that with other treatments, the yield was low. The yield treated with low concentration of B was 10% higher than that with high concentration, and the high concentration of B was not conducive to increase the content of the alkaloids. Among these treatments, The fourth treatment was the optimal combination, of which the volume of sub-roots was the largest and the most homogeneous, the growth of the vegetative organs was better and the accumulation of dry matters was more, the yield of this treatment was 10,754.7 kg x hm(-2), which was increased by 14.9%, and the content of alkaloid was increased by 13.9%. CONCLUSION: The ratio of 4 is the best treatment for high yield and quality cultivation of Aconite.

[Study on growth and dry substance accumulation of Curcuma longa].

OBJECTIVE: To explore the growth characteristics of Curcuma longa, and provide basis for standardized cultivation. METHOD: Plant samples were collected and investigated periodically. RESULT: According to the growth of different parts and the characteristics of dry substance accumulation of C. longa, the development of C. longa could be divided into five stages: emergence of seedlings, seedling, leaf, root tuber expansion, and dry substance accumulation of root tuber. In terms of number, leaf of C. longa increases gradually from one at first to eight at the final stage. Leaf size increases at a very low speed at the stage of seedling. However, leaves expands their sizes at a much higher speed at the stage of leaf. The dry substance in different parts accumulates increasingly with the development of C. longa dry substance mainly accumulates in leaves at the stage of leaf, and in rhizome at the stage of root tuber expansion. At the final stage, it mainly accumulates in root tuber. CONCLUSION: Cultivation technologies of C. longa and the relevant management methods could be established according to the growth of different parts of C. longa and the characteristics of dry substance accumulation in different stages.

[Effects of seeding and sowing methods on the yields of root tubers of Curcuma longa].

OBJECTIVE: To provide a scientific basis for standardizing the cultivation method for Curcuma longa. METHOD: Plant heights and seeding numbers were sampled periodically, the plot yields were counted a tharvested. RESULT: The effects of seeding method on yields, plant height and number were significant. The effects of different sowing ways on the yields were very little. CONCLUSION: The hole seeding method using mother tuber or blastostyle bearing three to four knots was shown to be the best may, worthy to be widely applied in production.