AIM:To conduct the relevance analysis of serum Dickkopf-1 (Dkk-1) and bone mineral density (BMD) in the different ages of female populations.METHODS: The women volunteers (n=100, 20~80 years old) were selected and divided into young group (20~39 years old), middle age group (40~59 years old) and elderly group (60~80 years old).The serum levels of Dkk-1 in the 3 groups of volunteers were detected by ELISA.In the middle age group, 25 people of 45~55 years old were selected and divided into postmenopausal group and premenopausal group to de-tect the serum level of Dkk-1 in the 2 groups of volunteers by ELISA.The BMD was measured by the method of dual energy X-ray absorptiometry.The differences of Dkk-1 expression levels among different ages of female populations, and the rele-vance with BMD were compared.RESULTS:With the increase in age, the serum Dkk-1 expression level increased ( P<0.05), and BMD were reduced (P<0.05).The blood level of Dkk-1 and BMD negatively correlated (P<0.05) in the 3 groups of volunteers.The serum levels of Dkk-1 and BMD had stronger negative correlation in postmenopausal women group than that in premenopausal women group.CONCLUSION:With the increase in age, the expression level of serum Dkk-1 increases and the BMD level decreases, which contribute to a risk of osteoporosis.In the same age range, the postmeno-pausal women express higher level of Dkk-1, and the decreased BMD is more obvious, which contribute to a greater risk of osteoporosis.The increased level of Dkk-1 also inhibits bone formation and promotes bone resorption.It may become a new target for preventing and treating osteoporosis.

The LEAFY (LFY) homologous gene of Dendrobium moniliforme (L.) Sw. was cloned by new primers which were designed based on the conservative region of known sequences of orchid LEAFY gene. Partial LFY homologous gene was cloned by common PCR, then we got the complete LFY homologous gene Den LFY by Tail-PCR. The complete sequence of DenLFY gene was 3 575 bp which contained three exons and two introns. Using BLAST method, comparison analysis among the exon of LFY homologous gene indicted that the DenLFY gene had high identity with orchids LFY homologous, including the related fragment of PhalLFY (84%) in Phalaenopsis hybrid cultivar, LFY homologous gene in Oncidium (90%) and in other orchid (over 80%). Using MP analysis, Dendrobium is found to be the sister to Oncidium and Phalaenopsis. Homologous analysis demonstrated that the C-terminal amino acids were highly conserved. When the exons and introns were separately considered, exons and the sequence of amino acid were good markers for the function research of DenLFY gene. The second intron can be used in authentication research of Dendrobium based on the length polymorphism between Dendrobium moniliforme and Dendrobium officinale.

The aim of this study is to investigate the therapeutic effect of RegIII-proinsulin-pBudCE4.1 plasmid on streptozotocin (STZ)-induced type 1 diabetes mellitus and its underlying mechanisms. The model of type 1 diabetes mellitus was established by intraperitoneal injections of STZ (40 mg kg(-1)) to Balb/c mice for five consecutive days. Then, ten type 1 diabetic mice were intramuscularly injected with 100 microg RegIII-proinsulin-pBudCE4.1 plasmid for 4 weeks (one time/week) and the blood glucose levels were monitored every week; whereas another ten diabetic mice served as negative control group were injected with pBudCE4.1 vector at the same dose. Normal control and model control mice were treated with normal saline at identical volume under the same way. Western blotting, MTT assay, ELISA, HE staining and Tunel assay were applied to explore the underlying mechanisms. Results showed that RegIII-proinsulin-pBudCE4.1 plasmid ameliorated the hyperglycemia symptoms in diabetic mouse remarkably. It induced an immunological tolerance state in type 1 diabetic mice by inhibiting the proliferation of splenic lymphocytes and recovering Th1/Th2 balance evidenced by MTT and ELISA analysis. Furthermore, it elevated insulin concentration in the serum of type 1 diabetic mice and promoted the regeneration of beta cells supported by the results of HE staining and Tunel assay. In conclusion, RegIII-proinsulin-pBudCE4.1 plasmid possesses powerful anti-diabetic ability, which may be involved in the inducing of immunological tolerance and enhancing beta cells recovery.

OBJECTIVETo explore the effects of moxibustion with seed-sized moxa cone at "Ganshu" (BL 18) on liver furiction and morphology in rat with precancerous lesion of hepatic cellular cancer MCC).

METHODSA total of 60 male Wistar rats were randomly divided into a normal group (10 rats), a model group (20 rats), a 20-day treatment group (15 rats) and a 40-day treatment group (15 rats). HCC model was established by intraperitoneal injection of diethylnitrosamine (DEN). Rats in the normal group received no treatment. Rats in the model group were treated with fixation. Rats in the 20-day treatment group and 40-day treatment group were treated by moxibustion with seed-sized moxa cone at "Ganshu" (BL 18), once every other day, for 20 days and 40 days, respectively. Blood sample in each group was collected 1 d before model establishment, 20 d, 40 d and 84 d after model establishment. Chemical method was applied to test the activity of ALT (alamine aminotransferase), AST (aspartate transaminase) and GGT (glutamyl transpeptidase); at the end of model establishment, all the rats were sacrificed to observe the liver morphology changes.

RESULTSAfter the first therapeutic course, the. content of ALT and AST in the 20-day treatment group was significantly lower than that in the model group (all P<0. 05); after the second therapeutic course, the content of ALT, AST and GGT in the 40-day treatment group was insignificantly lower than that in the model group (all P>0. 05). Under light microscope, the slice of liver tissue indicated that primary tumor was induced in the model group, and the tumor cells were stained and irregular; the cytoplasm in the 20-day treatment group was even, and the tumor cells were few with several nodules alone. In the 40-day treatment group the liver morphology was normal and the staining was even; the tumor cells were few without nodules or a few. Conclusion Moxibustion with seed-sized moxa cone at "Ganshu" (BL 18) could reduce the serum content of ALT, AST and GGT in rats with HCC, which could protect the liver and: delay the DEN-induced precancerous lesion on some levels.

Acupuncture Points ; Animals ; Aspartate Aminotransferases ; blood ; Humans ; Liver ; pathology ; Liver Neoplasms ; enzymology ; pathology ; therapy ; Male ; Moxibustion ; Rats ; Rats, Wistar ; gamma-Glutamyltransferase ; blood

This study examined the potential antilithic effects of a traditional Chinese medicine Urtica dentata Hand (UDH) in experimental rats and screened the optimal extract of UDH as a possible therapeutic agent for kidney stones. The rat model of urinary calcium oxalate stones was induced by intragastric (i.g.) administration of 2 mL of 1.25% ethylene glycol (EG) and 1% ammonium chloride (AC) for 28 days and was confirmed by Color Doppler ultrasound imaging. The rats in different experimental groups were then intragastrically given petroleum ether extract (PEE), N-butanol extract (NBE), aqueous extract (AqE) of UDH, Jieshitong (positive control drug), and saline, respectively. Treatment with NBE significantly reduced the elevated levels of urinary calcium, uric acid, phosphate, as well as increased urinary output. Accordingly, the increased calcium, oxalate levels and the number of calcium oxalate crystals deposits were remarkably reverted in the renal tissue of NBE-treated rats. In addition, NBE also prevented the impairment of renal function to decrease the contents of blood urea nitrogen (BUN) and creatinine. Taken together, these data suggest that NBE of UDH has a beneficial effect on calcium oxalate urinary stones in rats by flushing the stones out and protecting renal function.

Objective To exploring the mechanism of Jinshui Chenfei formula(JCF)in ameliorating silica(SiO2)-induced silicosis fibrosis based on endogenous metabolite changes.Methods A total of 32 SPF male Sprague-Dawley(SD)rats were divided into normal control group,model group,JCF group(9.72 g·kg-1·d-1),and Tetrandrine group(27 mg·kg-1·d-1)according to random number table method.The experimental silicosis model was established by intratracheal injection with SiO2 suspension(250 mg/kg)on day 1.From week 5-8,silicosis rats were treated with tetrandrine or JCF.On the end of week 8,the changes of pulmonary function index,including forced vital capacity(FVC),tidal volume(TV)and lung dynamic compliance(Cydn)were detected.The pathological changes of lung tissue were analyzed by hematoxyline-osin(HE)staining and Masson staining,the severity of focal alveolitis and fibrosis was also evaluated using the Szapiel scale and the Ashcroft scale,the positive staining of collagen Ⅰ(COL Ⅰ)and COL Ⅲ was detected using immunohistochemistry;the protein expression of transforming growth factor-β1(TGF-β1),fibronectin(FN),andα-smooth muscle actin(α-SMA)were measured by Western blotting.The rat serum samples were further screened for differential metabolites using ultra performance liquid chromatographytandem quadrupole time of flight mass spectrometr(UPLC-Q-TOF-MS)and pathway analysis was performed based on MetaboAnalyst 5.0.Results Compared with those in the normal control group,pathological changes such as alveolar structure destruction,the fibrous nodules encapsulated SiO2 particles were increased in lung tissues of rats in model group,alveolitis score and pulmonary fibrosis score were significantly higher(alveolitis score:2.62±0.27 vs.0.20±0.15,pulmonary fibrosis score:5.42±0.66 vs.0.50±0.84,both P<0.01);pulmonary function index including Cydn,FVC,and TV were significantly decreased[Cdyn(mL/cmH2O):0.26±0.03 vs.0.33±0.03,FVC(mL):8.09±0.47 vs.10.99±0.38,TV(mL):1.95±0.19 vs.2.53±0.26,all P<0.01];positive staining of COL Ⅰ,COL Ⅲ and ɑ-SMA,FN,TGF-β1 proteins expression showed higher in lung tissues[positive staining of COL Ⅰ(A value):13.47±1.76 vs.5.77±0.45;positive staining of COL Ⅲ(A value):10.39±0.47 vs.6.19±0.77,FN protein expression(FN/GAPDH):0.33±0.02 vs.0.21±0.07,α-SMA protein expression(α-SMA/GAPDH):1.78±0.16 vs.1.11±0.24,TGF-β1 protein expression(TGF-β1/GAPDH):0.52±0.10 vs.0.11±0.46,all P<0.01].Compared with the model group,the pathological changes of lung tissues were almost restored,alveolitis score and lung fibrosis score were significantly reduced in JCF and Tetrandrine groups(alveolitis score:1.10±0.15,1.33±0.31 vs.2.62±0.27,pulmonary fibrosis score:3.50±0.45,4.33±0.98 vs.5.42±0.66,all P<0.01);the pulmonary function index Cydn,FVC and TV were significantly increased[Cdyn(mL/cmH2O):0.32±0.05,0.31±0.04 vs.0.26±0.03,FVC(mL):9.41±0.85,8.70±0.92 vs.8.09±0.47,TV(mL):2.70±0.19,2.27±0.15 vs.1.95±0.19,all P<0.05];positive staining of COL Ⅰ,COL Ⅲ,and protein expression of FN,ɑ-SMA,and TGF-β1 in lung tissues was significantly decreased[COL Ⅰ(A value):7.09±0.67,8.13±0.64 vs.13.47±1.76,COL Ⅲ(A value):8.19±0.66,8.52±0.22 vs.10.39±0.47,FN protein expression(FN/GAPDH):0.19±0.06,0.24±0.03 vs.0.33±0.02,α-SMA protein expression(α-SMA/GAPDH):0.89±0.41,0.88±0.08 vs.1.78±0.16,TGF-β1 protein expression(TGF-β1/GAPDH):0.04±0.03,0.06±0.01 vs.0.52±0.10,all P<0.05].Metabolomics analysis showed that a total of 10 major differential metabolites were identified between normal control group,model group and JCF group,including arachidonic acid,palmitic acid,indole-3-acetic acid,propionylcarnitine,(S)-4-hydroxymandelonitrile,nalidixic acid,benzocaine,gramine,4-ethylphenol,N-benzylfor mamide.The differential metabolites in silicosis rats reversed by JCF treatment were mainly enriched,including unsaturated fatty acid biosynthesis,arachidonic acid metabolism,tryptophan metabolism,fatty acid elongation,fatty acid degradation and biosynthesis.Conclusion JCF could effectively improve the silicosis fibrosis,which is mainly related to biosynthesis of unsaturated fatty acids biosynthesis,arachidonic acid metabolism,tryptophan metabolism,fatty acid elongation,fatty acid degradation and biosynthesis.

Objective To analyze the clinical characteristics of lung adenocarcinoma patients with positive EGFR mutations detected in pleural effusion.Methods We retrospectively analyzed the clinical characteristics including gender,age,smoking history,presence of other underlying diseases(such as COPD,cardiovascular disease,and diabetes),site of pleural fluid,feature of pleural fluid,and TNM stage in patients with lung adenocar-cinoma who had been admitted to the first Affiliated Hospital of Bengbu Medical College from 2020.01 to 2022.12 for the first time by the detection of EGFR mutation positive in pleural effusion.The data were statistically analyzed using the SPSS 26.0 software.Results A total of 126 patients were screened for enrollment,including 61 patients(48.41%)with EGFR exon 19 deletion mutation(19del),56 patients(44.44%)with exon 21 L858R mutation(21L858R),and 9 patients(7.14%)with non-classical mutations.Univariate analysis showed that the three muta-tion subtypes were statistically significant in terms of gender,age,smoking history,and presence of COPD(P<0.05 for all comparisons),but not in terms of pleural fluid site,feature of pleural fluid,tumor size,and presence of cardiovascular disease,diabetes mellitus,presence of distant metastases,and mediastinal lymph node metastases(P>0.05 for all comparisons);Multivariate analysis showed that 21 L858R mutation was more likely to be found in male,older age,non-smoking,and presence of COPD than 19del mutation;non-classical mutation was more likely to be found in male than 19del mutation.Conclusions There are significant differences among the three mutation subtypes in sex,age,smoking history,and presence of COPD,but not in pleural fluid location,feature of pleural fluid,tumor size,presence of cardiovascular disease or diabetes mellitus,presence of distant metastases,or medias-tinal lymph node metastases;Among lung adenocarcinoma patients with positive EGFR mutations in pleural fluid,21 L858R mutation mostly occurs in male,older age,non-smokers,and those complicated with COPD,while non-classical mutation mainly develops in male.However,more case studies are needed to confirm the above conclusions.

To explore the application of gene chip technology in the acupuncture research so as to provide evidences for the mechanism of acupuncture for regulating bodies. The literature on the application of gene chip technology in the acupuncture field from 2001 to 2016 was collected in PubMed, Springer, CNKI and WANFANG databases, which was analyzed and summarized. There were some achievements of the technology for acupuncture research, focusing on the five aspects, including the study of the relationship between meridian-point and viscera, the influencing factors of acupuncture effect, the effect and mechanism of acupuncture analgesia, the mechanism of acupuncture anti-aging, the effect and mechanism of acupuncture for diseases of each system. Gene chip technology plays an important role in researching acupuncture mechanism. It is an important technology for genomics study of acupuncture. However, there are also some disadvantages such as high cost, deficient data mining, non-uniform observation objects, deficient professionals, etc. All those need further resolution so as to promote the application of this technology in the acupuncture researching field.