AIM To prepare curcumin-loaded poly (D,L-lactide-co-glycolide) acid (PLGA) nanoparticles,and to investigate their effects on promoting wound healing.METHODS Phacoemulsification method was adopted in the preparation of PLGA nanoparticles.With lactic acid-glycolic acid ratio,surfactant type and concentration and PLGA consumption as influencing factors,encapsulation efficiency,drug loading,particle size and PDI as evaluation indices,the preparation process was optimized by single factor test.The morphology was observed under transmission electron microscopy,after which the in vitro drug release was determined by dialysis bag method,the optical properties were investigated by fluorescence spectroscopy,and the effects on promoting wound healing of mouse skin.RESULTS The optimal conditions were determined to be 50∶50 for lactic acid-glycolic acid ratio,60 mg for PLGA consumption,and 3％ for surfactant (PVA) concentration.The obtained PLGA nanoparticles with spherical core-shell structure demonstrated the average particle size,PDI,Zeta potential and accumulative release rate within 168 h of (149.70±7.27) nm,0.28±0.04 and (-10.18±1.00) mV and (39.21±1.44)％,respectively,which displayed typical aggregation-induced emission property,excitation wavelength-dependent property and optical stability.Compared with the PBS group,PBS+illumination group,curcumin+illumination group and preparation group,the preparation+illumination group exhibited increased wound healing rate (P<0.05,P<0.01) and reduced wound epidermal thickness (P<0.05,P<0.01).CONCLUSION Curcumin-loaded PLGA nanoparticles show good optical properties,which can promote the wound healing of mouse skin when combined with photodynamic therapy.

Objective:To evaluate the effect of five-element acupuncture on clinical symptoms, brain metabolism and immunoglobulin level in patients with chronic fatigue syndrome.Methods:It was a randomized controlled trial. A total of 94 patients with chronic fatigue syndrome in our hospital from January 2021 to January 2022 were selected and divided into two groups according to the random number table method, with 47 in each group. The conventional western medicine group was treated with conventional western medicine, and the five-element acupuncture group was treated with five-element acupuncture on the basis of the conventional western medicine group. Both groups were treated for 4 weeks. Before and after treatment, the serum levels of interferon-γ (IFN-γ), corticosterone (CORT), IL-2 and 5-hydroxytryptamine (5-HT) were detected by ELISA; the levels of natural killer (NK) cells, CD4 +, CD8 +, IgG and IgM were detected by flow cytometry; the whole body superconducting MRI scanner was used to scan T2 Flair, T2WI and TlWI sequences of the hippocampus, and the spectral curves and the areas under the peak of N-acetylaspartic acid (NAA), creatine (Cr) and choline (Cho) were obtained, and the ratios of Cho/Cr and NAA/Cr were calculated. the fatigue Scale-14 (FS-14) and Fatigue Severity Scale (FSS) were used to evaluate the fatigue state of the patients, and the Depression-Anxiety-Stress Scale-21 (DASS-21) and Beck Anxiety Inventory (BAI) were used to evaluate the anxiety state of the patients. Symptom Checklist 90 (SCL-90) and Somatic and Mental Health Report Score (SPHERE) were used to evaluate the quality of life of patients. And the clinical efficacy was evaluated. Results:After treatment, the levels of IgG, CD4 + and NK in the five-element acupuncture group were significantly higher than those in the conventional western medicine group ( t values were 4.76, 3.65, 6.42, respectively, P<0.01), and the level of IgM, CD8 + was significantly lower than that in the conventional western medicine group ( t values were 7.30, 4.79, P<0.01); the levels of IFN-γ, IL-2 and 5-HT in the observation group were significantly higher than those in the conventional western medicine group ( t values were 7.60, 4.05, 2.79, respectively, respectively, P<0.01), and the level of CORT was significantly lower than that in the conventional western medicine group ( t=6.72, P<0.01); the NAA/Cr levels in the left [(1.10±0.04) vs. (1.05±0.03), t=6.86] and right [(1.18±0.02) vs. (1.21±0.03), t=8.23] hippocampus of the experimental group were significantly higher than those in the conventional western medicine group ( P<0.01), and the Cho/Cr levels in the left [(1.08±0.04) vs. (1.03±0.03), t=5.70] and right [(1.17±0.02) vs. (1.20±0.03), t=5.71] hippocampus of the experimental group were significantly lower than those of the conventional western medicine group ( P<0.01). After treatment, the scores of physical fatigue, mental fatigue and FSS in the five-element acupuncture group were significantly lower than those in the conventional western medicine group ( t values were 8.08, 9.08 and 7.07, respectively, P<0.01). The scores of DASS-21, BAI, SCL-90 and SPHERE in the conventional western medicine group were significantly lower than those in the conventional western medicine group ( t values were 3.63, 5.77, 8.74, 5.92, respectively, P<0.01).The total effective rate was 95.74% (45/47) in the five-element acupuncture group and 82.98% (39/47) in the conventional western medicine group, and there was no significant difference between the two groups ( χ2=2.80, P=0.094). Conclusion:Five-elements acupuncture can improve the expression of T lymphocytes, increase the levels of immunoglobulin and NK, reduce the level of CORT, regulate the brain metabolism of NAA in the left and right hippocampus, improve the clinical symptoms and negative emotions, and improve the clinical efficacy and quality of life in patients with chronic fatigue syndrome.

ObjectiveTo investigate the possible mechanism of electroacupuncture at "Neiguan" （PC6） and "Zhongwan" （RN12） in the treatment of functional dyspepsia （FD）. MethodsSPF male SD rats were randomly divided into model group， Neiguan （PC6） group， Zhongwan （RN12） group， and Neiguan-Zhongwan （PC6-RN12） group， with 8 rats in each group. Except for the normal group， the other groups of rats were cogavaged with 0.1% sucrose iodoacetamide solution combined with small platform standing training to establish FD rat models. After successful modeling， the rats in the normal group and model group were tied up for 30 min/d for 7 days； the Neiguan group， Zhongwan group， and Neiguan-Zhongwan group were treated with electroacupuncture intervention at "Neiguan" （PC6）， "Zhongwan" （RN12）， and "Neiguan" - "Zhongwan" （PC6-RN12） acupoints， respectively， using continuous wave， frequency of 2 Hz， current intensity of 1 mA， 30 min/d， and continuous intervention for 7 days. The general condition of rats in each group was observed. After treatment， the body weight and food intake of rats were measured， and the gastric emptying rate was calculated； HE staining was performed on the gastric antrum tissue of rats to observe the histopathologic changes； the expressions of calcitonin gene-related peptide （CGRP） and Ghrelin protein in gastric antrum were detected by Western Blot； the metabolites in gastric antrum tissues were detected by liquid chromatography-mass spectrometry （LC-MS）， and differential metabolites were screened by correlation analysis， then Metabo Analyst 5.0 and KEGG databases were used for metabolic pathway analysis. ResultsUnder light microscope， the gastric antrum structure was complete and the glands were abundant. No obvious inflammation and edema were found in gastric mucosa. Compared with the normal group， the body weight， food intake， and gastric emptie rate of rats in model group decreased， the expression of Ghrelin protein decreased and the expression of CGRP protein increased in gastric antrum tissue （P＜0.01）. Compared with model group， the body weight， food intake， and gastric emptance rate of rats in Neiguan group， Zhongwan group and Neiguan-Zhongwan group all increased， CGRP protein expression decreased in Neiguan group， and Ghrelin protein expression increased and CGRP protein expression decreased in Zhongwan group and Neiguan-Zhongwan group （P＜0.01 or P＜0.05）. Compared with Neiguan-Zhongwan group， Ghrelin protein expression decreased and CGRP protein expression increased in Neiguan group and Zhongwan group （P＜0.05 or P＜0.01）. Metabolomics results showed that compared with normal group， the content of metabolites adenosine diphosphate ribose， adenosine monophosphate and adenosine diphosphate in gastric antrum tissue of model group decreased； compared with model group， the contents of adenosine diphosphate， adenosine diphosphate and citicoline in Neiguan group increased， the contents of nicotine adenine dinucleotide， cytidine diphosphate ethanolamine and citicoline in Zhongwan group increased， and the contents of adenosine diphosphate， cytidine diphosphate and citicoline in Neiguan-Zhongwan group increased （P＜0.05 or P＜0.01）. Compared with model group， the main metabolic pathways of different metabolites in PC6-RN12 group were glycerophospholipid metabolism， taurine and hypotaurine metabolism， purine metabolism， starch and sucrose metabolism. ConclusionElectroacupuncture at “Neiguan” and “Zhongwan” acupoints can effectively regulate gastrointestinal motility and improve FD symptoms in FD rats， and the effect is better than that of "Neiguan" or "Zhongwan" acupoints alone. The mechanism may be related to the influence of related metabolites on energy metabolism， glucose metabolism and nucleotide metabolism， thereby regulating gastrointestinal motility hormones.

AIM To prepare curcumin-loaded poly (D,L-lactide-co-glycolide) acid (PLGA) nanoparticles,and to investigate their effects on promoting wound healing.METHODS Phacoemulsification method was adopted in the preparation of PLGA nanoparticles.With lactic acid-glycolic acid ratio,surfactant type and concentration and PLGA consumption as influencing factors,encapsulation efficiency,drug loading,particle size and PDI as evaluation indices,the preparation process was optimized by single factor test.The morphology was observed under transmission electron microscopy,after which the in vitro drug release was determined by dialysis bag method,the optical properties were investigated by fluorescence spectroscopy,and the effects on promoting wound healing of mouse skin.RESULTS The optimal conditions were determined to be 50∶50 for lactic acid-glycolic acid ratio,60 mg for PLGA consumption,and 3％ for surfactant (PVA) concentration.The obtained PLGA nanoparticles with spherical core-shell structure demonstrated the average particle size,PDI,Zeta potential and accumulative release rate within 168 h of (149.70±7.27) nm,0.28±0.04 and (-10.18±1.00) mV and (39.21±1.44)％,respectively,which displayed typical aggregation-induced emission property,excitation wavelength-dependent property and optical stability.Compared with the PBS group,PBS+illumination group,curcumin+illumination group and preparation group,the preparation+illumination group exhibited increased wound healing rate (P<0.05,P<0.01) and reduced wound epidermal thickness (P<0.05,P<0.01).CONCLUSION Curcumin-loaded PLGA nanoparticles show good optical properties,which can promote the wound healing of mouse skin when combined with photodynamic therapy.

OBJECTIVES: To observe the effects of electroacupuncture (EA) at "Neiguan" (PC 6) and "Zusanli"(ST 36) on the gastric emptying rate, the level of serotonin (5-HT) and the protein expression of motilin (MTL), ghrelin, substance P (SP) and vasoactive intestinal peptide (VIP) in the antral tissue of the rats with functional dyspepsia (FD) and explore the effect mechanism of EA in treatment of FD. METHODS: A total of 21 SPF male SD rat pups were randomly divided into a normal group, a model group and an EA group, with 7 rats in each group. In the model group and the EA group, FD model was prepared by the gavage with 0.1% sucrose iodoacetamide solution combined with the modified small platform method. After the successful modeling, EA was applied to "Neiguan" (PC 6) and "Zusanli"(ST 36) in the rats of the EA group, with disperse-dense wave, 20 Hz/100 Hz in frequency, stimulated for 30 min, once daily, for 7 days consecutively. Before and after intervention, the general condition of the rats was observed in each group. After the completion of intervention, the gastric emptying rate was measured, the morphological changes of gastric antral tissue were observed using HE staining, the level of 5-HT was detected with ELISA method, and the protein expression of MTL, ghrelin, SP, and VIP was determined with Western blot method in the antral tissue of rats. RESULTS: In the normal group, the rats were in a good mental state, with lustrous fur, flexible movement and the increase of food intake and body mass. In the model group, the rats were poor in mental state, lack of lustre in fur, preference for the body curled up, reduced activity and response; and a part of rats had loose stool, obviously enlarged gastric body and gastric food retention. In the EA group, the general condition of rats, e.g. the mental state, food intake and activity, were improved, the gastric body got smaller obviously and the gastric food retention was reduced when compared with the model group. The antral structure was intact, the glands were rich and no injury of the gastric mucosa was found, e.g. inflammatory reaction and edema in the rats of each group. Compared with the normal group, the gastric emptying rate was decreased (P<0.01), 5-HT level was increased (P<0.01), the protein expression of MTL and ghrelin was reduced (P<0.01) and that of VIP was elevated (P<0.01) in the rats of the model group. The gastric emptying rate was increased (P<0.01), 5-HT level was decreased (P<0.01), and the protein expression of MTL and ghrelin was elevated (P<0.05, P<0.01) in the rats of the EA group when compared with those in the model group. CONCLUSIONS Electroacupuncture at "Neiguan" (PC 6) and "Zusanli"(ST 36) may effectively relieve gastric dysfunction, strengthen gastric motility and promote gastric emptying so as to alleviate the symptoms of dyspepsia in FD rats, and its mechanism may be related to the regulation of gastrointestinal hormones in the antral tissue.
Rats ; Male ; Animals ; Electroacupuncture ; Dyspepsia/therapy* ; Rats, Sprague-Dawley ; Ghrelin ; Gastrointestinal Hormones ; Serotonin ; Vasoactive Intestinal Peptide ; Acupuncture Points

OBJECTIVE: To explore the pathogenic variants and clinical classification of two fetuses with Short-rib thoracic dysplasia with or without polydactyly (SRTD). METHODS: With informed consent obtained, the phenotypic characteristics of the fetuses were comprehensively examined, and genomic DNA was extracted from fetal skin tissue and peripheral blood samples of the parents with conventional phenol-chloroform method. Whole exome sequencing (WES) was carried out on both fetuses, and the candidate variants were validated by Sanger sequencing. The pathogenicity of the candidate variants was analyzed using bioinformatic software VarCards, and the impact of the variants on the protein structure was predicted with Swiss-Pdb-viewer. RESULTS: Both fetuses were found to harbor compound heterozygous variants of the DYNC2H1 gene, including c.515C>A (p.Pro172Gln) and c.5983G>A (p.Ala1995Thr) in fetus 1, and c.5920G>T (pGly1974) and c.9908T>C (p.He3303Thr) in fetus 2. The parents of both fetuses were heterozygous carriers. CONCLUSION The compound heterozygous variants of the DYNC2H1 gene probably underlay the SRTD3 in the two fetuses.
Humans ; Fetus ; Chloroform ; Computational Biology ; Ethnicity ; Ribs

OBJECTIVE: To explore the genetic basis for three Chinese patients with McCune-Albright syndrome (MAS). METHODS: Three children who had respectively presented at Shandong Provincial Hospital in April 2019 and Peking Union Medical College Hospital in August 2020 and May 2021 were selected as the research subjects. Peripheral blood samples of the probands and their family members were taken for the extraction of genomic DNA. Potential variants were screened by whole exome sequencing (WES), and candidate variants were validated by Sanger sequencing of the patients and their family members. RESULTS: The proband from family 1 was found to harbor a heterozygous c.601C>T (p.R201C) missense variant in exon 8 of the GNAS gene, whilst the probands from families 2 and 3 were both found to harbor a heterozygous c.602G>A (p.R201H) missense variant in exon 8 of the GNAS gene. Both variants were known to be pathogenic, and all probands were found to be mosaics for the corresponding variants but with various degrees. CONSLUSION WES can effectively diagnose MAS and other somatic genetic disorders. In this study, the combined WES and Sanger sequencing have verified the degree of mosaicisms of pathogenic variants in the three MAS patients, albeit no apparent correlation was found between the degree of mosaicisms and the phenotype of patients. Above finding has provided a basis for genetic counseling and prenatal diagnosis for the affected families.
Humans ; Mutation ; Fibrous Dysplasia, Polyostotic/genetics* ; East Asian People ; Exons ; Phenotype ; Pedigree

Objective To evaluate the effects of early exercise intervention in patients who have undergone primary isolated valve surgery. Methods Forty patients scheduled for mitral, aortic, and/or tricuspid valve surgery were allocated to receive a supervised exercise intervention consisting of cycling for 3 min/d at the bedside after operation (intervention group, n=20, mean age [49.05±3.728] years) or to receive no exercise intervention (control group, n=20, mean age [47.95 ± 3.214] years). Oxygen saturation (SpO2) was measured by pulse oximetry continuously before and after the 6-minute walk test. Psycho-educational counseling was provided, and patients were assessed using standard patient questionnaires. Results The arterial SpO2 level increased significantly in the intervention group after exercise compared with the control group (P<0.05). Heart rate returned to baseline in the intervention group postoperatively and was significantly lower than that in the control group (P<0.05). Conclusions A small amount of supervised cycling exercise at the bedside is a safe activity that may improve peripheral arterial SpO2 and reduce heart rate to the baseline level following longer distance walk before discharge in patients who have undergone isolated valve surgery.

Objective:To identify pathogenicity of the potential splicing variants in two Chinese Han patients with osteogenesis imperfecta.Methods:Genomic DNA was extracted using the conventional phenol-chloroform method; whole exome sequencing (WES) was used to analysis the disease-related variants in the two probands; Minigene assay was used to identify pathogenicity of the variants found in the patients' genome that possibly affect RNA splicing.Results:Two potential splicing variants, c.858+1_858+5delGTAAG in intron 12 of COL1A1 and c.1405-7C>T in intron 24 of COL1A2, were found in proband 1 and proband 2, respectively. In addition, a missense mutation, c.2972G>T (p.G991V) in exon 45 of COL1A2, was detected in proband 2. Minigene assay revealed that the variant in proband 1 caused the skipping of exon 12, while the variant in proband 2 did not lead to aberrant splicing. G199 of the COL1A2 in proband 2 was a highly conserved amino acid site, and the results suggested that c.2972G>T (p.G991V) may be the real pathogenic variant by the means of bioinformatics analysis.Conclusion:The variant c.858+1_858+5delGTAAG in COL1A1 was a causative variant that led to OI in proband 1, while the missense variant c.2972G>T (p.G991V) in COL1A2 was the cause of OI in proband 2, instead of the variant c.1405-7C>T. Minigene assay for potential splicing variants detected by WES could not only validate the pathogenicity of the candidate variants and enrich the mutation spectrum of OI, but also lay the foundation for patients' prenatal diagnosis and subsequent mechanism research.

On the base of the idea of traditional Chinese medicine as "disease prevention", the mode and the protocol of the moxibustion intervention for the group under quarantine after close contact with coronavirus disease 2019 (COVID-19) were explored. The group under quarantine after close contact with COVID-19 was taken as the subjects. By the non-contact physician-patient communication network platform co-developed by China Association of Acupuncture-Moxibustion, Hunan Provincial Association of Acupuncture-Moxibustion, Data Center of China Academy of Chinese Medical Sciences and Yuge Medicine Company, an exploratory randomized controlled trial was designed. A total of 100 cases were included and randomized into a moxibustion group and a conventional intervention group, 50 cases in each one. In the moxibustion group, moxibustion intervention was used. In the conventional intervention group, the conventional observation was adopted without moxibusiton intervention applied. The outcomes included the symptoms changes, e.g. anxiety, emotional disturbance, fatigue, headache and diarrhea, as well as whether quarantine release and the case confirmed or not, etc. The results were evaluated before intervention, in 14 days of intervention and 2 weeks after intervention separately. In this research, on the base of internet plus technology and with the internet communication platform adopted, through mobile phone WeChat App, it was to implement the subject screen, the random allocation and the instruction of moxibustion intervention as well as the quality control of patient's diary and data collection. It is anticipated that the significance and the implementation mode of moxibustion intervention can be assessed preliminarily for the group under quarantine after close contact with COVID-19.
Acupuncture Therapy ; methods ; Betacoronavirus ; China ; Coronavirus Infections ; therapy ; Humans ; Moxibustion ; Pandemics ; Pneumonia, Viral ; therapy ; Quarantine