Aim: To improve the solubility, dissolution and bioavailability of lycopene by preparing lycopene solid dispersion. Methods: Lycopene solid dispersion was prepared by the solvent method using Poloxamer 188 as car-rier. The physicochemical characteristics of the dispersion were determined by DSC, ultraviolet-visible spectra and the Dissolution Apparatus Ⅱ( Paddle). The oral bioavailability of lycopene was estimated in rat after oral dosing of lycopene solid dispersion or oil preparation. The plasma concentrations of lycopene in rats were determined by HPLC. The pharmacokinetic parameters were estimated by Kinetica software package. Results: In vitro dissolution of lycopene solid dispersion was greater than those of lycopene (raw material), and the physical mixture of lyco-pene and Poloxamer 188, partly due to the existing molecular state of lycopene in the dispersion. It was also found that the relative bioavailability of lycopene solid dispersion to lycopene oil preparation was (312. 2±96. 9) % . The optimal ratio of lycopene to carrier in the dispersion was about 1: 5. Conclusion: Lycopene-Poloxamer 188 solid dispersion could be prepared by the proposed simple, low-costly procedure resulting in improved bioavail-ability of lycopene, which is worthy of further development.

Objective To estabolish the ALDH2 Glu504Lys genotyping technique for oral swab sample type based on PCR-gold magnetic nanoparticle chromatographyl.Methods According to the ALDH2 Glu504Lys gene the specific primers were designed by ARMS-PCR technique.Optimized the optimal ALDH2 Glu504Lys oral swab reaction system and PCR reaction procedures based on the routine reaction conditions and reaction procedure of laboratory PCR.60 samples of oral swabs were collected and tested by gold magnetic nano-chromatography.The results were verified by sequencing.Results The detected 60 cases of oral swab samples contain 16 cases heterozygous mutation,wild type 42 cases,homozygous mutation 2 cases and then the results by Goldmag nanoparticle detection was perfectly matched with sequencing results.Conclusion The Goldmag nanoparticles chromatographic detection technique of ALDH2 Glu504Lys buccal swabs system was established.With accuracy,fastness,reliability,it is worthy of clinical promotion.

For the purposes of meeting the needs of COPD patients for health education, decreasing times and costs of re-hospitalization, and increasing patient satisfaction and nurses' working enthusiasm. In 2011, our hospital established a specialist nursing clinic for chronic obstructive pulmonary disease (COPD), and two nursesentitled-deputy directorswith rich clinical experiences for more than fifteen years and strong communication skills were se-lected to take charge of the clinic, and they were responsible for nursing assessment, making individualized pul-monary rehabilitation plan, health guidance, long-term follow up, etc., for discharged stable COPD patients and COPD patients with no need for hospitalization. Patient satisfaction was improved from 83% to 96% after implemen-tation.

Objective: To explore the possible mechanism of moxibustion in myocardial protection of rats undergoing long-term fatigue exercise based on observing the classical pyroptosis pathway mediated by nuclear factor kappa-B (NF-κB)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/cysteinyl aspartate specific proteinase 1 (Caspase-1).Methods: A total of 50 specific-pathogen-free male Sprague-Dawley rats were bought. Ten unqualified rats were excluded, and the remaining 40 rats were divided into a normal group, a normal + Shenque (CV8) group, a model group, a model + non-meridian non-point group, and a model + Shenque (CV8) group according to the random number table method, with 8 rats in each group. Except for rats in the normal group and the normal + Shenque (CV8) group, rats in the other three groups were trained with an incline running table exercise protocol to create a long-term fatigue exercise model, 1 h/time, once a day for 5 d with 2 d off, for a total of 8 weeks. Rats in the normal group received no modeling or intervention. Rats in the normal + Shenque (CV8) group were not modeled but received mild moxibustion at Shenque (CV8); those in the model group were modeled only without intervention; those in the model + non-meridian non-point group received moxibustion at non-meridian and non-point spots after the modeling; those in the model + Shenque (CV8) group received moxibustion at Shenque (CV8) after modeling. The above moxibustion interventions were performed for 15 min/time once daily, for 5 d with 2 d off per week and a total of 8 weeks. Blood was collected from the femoral artery 4 h after the last exercise, and the serum interleukin (IL)-1β and IL-18 levels were measured. The NF-κB, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, and gasdermin D (GSDMD) expression levels were detected by Western blotting. Myocardial morphology and pyroptosis were observed by hematoxylin-eosin (HE) staining and electron microscopy. Results: The HE staining results showed that the myocardial cells in the model group and the model + non-meridian non-point group were disorganized with blurred transverse lines, widened interstitial spaces, interstitial edema, and inflammatory cell infiltration. The structure of myocardial cells in the model + Shenque (CV8) group was clearly visible, with slightly widened interstitial spaces and occasional infiltration of inflammatory cells in the interstitium. Compared with the normal group, the serum IL-1β and IL-18 levels were increased, and myocardial NF-κB, NLRP3, ASC, Caspase-1, and GSDMD expression levels were elevated in the model group and the model + non-meridian non-point group (P<0.01). Compared with the model group, the above indicators did not change significantly in the model + non-meridian non-point group, while all the above indicators were decreased in the model + Shenque (CV8) group (P<0.01). Compared with the model + non-meridian non-point group, all the above biochemical indicators were decreased in the model + Shenque (CV8) group (P<0.01). Transmission electron microscopy showed that the mitochondria number was increased in the model group and the model + non-meridian non-point group, some of the mitochondrial lumen was irregularly enlarged, the cell membrane structure was unclear, and chromatin was aggregated. The mitochondria number was increased, the swelling was reduced, and the nuclear membrane structure was more intact in the model + Shenque (CV8) group. Conclusion: Moxibustion at Shenque (CV8) regulates the NF-κB/NLRP3/Caspase-1 pathway and reduces the pyroptosisin the myocardium of rats with long-term fatigue exercise, thus reducing the myocardial injury caused by long-term fatigue exercise.

This study aimed to explore the expression changes of VASA gene in sheep testis development and to construct VASA gene knock-in vector to prepare for the study on the differentiation of sheep germ cells in vitro. The testicular tissues of 3-month-old (3M) and 9-month-old (9M) sheep which represent immature and mature stages, respectively, were collected. The differential expression of VASA gene was analyzed by quantitative real-time PCR (qPCR) and Western blotting, and the location of VASA gene was detected by immunohistochemistry. The sgRNA targeting the VASA gene was designed and homologous recombination vectors were constructed by PCR. Subsequently, plasmids were transferred into sheep ear fibroblasts. The VASA gene was activated in combination with CRISPR/dCas9 technology to further verify the efficiency of the vector. The results showed that the expression level of VASA gene increased significantly with the development of sheep testis (P < 0.01), and was mainly located in spermatocytes and round spermatids. The knock-in vector of VASA gene was constructed by CRISPR/Cas9 system, and the Cas9-gRNA vector and pEGFP-PGK puro-VASA vector were transfected into ear fibroblasts. After CRISPR/dCas9 system was activated, ear fibroblasts successfully expressed VASA gene. The results suggest that VASA gene plays a potential function in sheep testicular development and spermatogenesis, and the VASA gene knock-in vector can be constructed in vitro through the CRISPR/Cas9 system. Our results provided effective research tools for further research of germ cell development and differentiation.
Male ; Animals ; Sheep/genetics* ; CRISPR-Cas Systems/genetics* ; Gene Knock-In Techniques ; RNA, Guide, CRISPR-Cas Systems ; Plasmids ; Germ Cells