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1.
Chinese Journal of Urology ; (12): 339-343, 2021.
Article in Chinese | WPRIM | ID: wpr-885018

ABSTRACT

Objective:To evaluate the best parameter of predicting the operation time and clearance of flexible ureteroscopic lithotripsy through comparing correlations between three stone burden parameters (diameter, area, volume) and the operation time or clearance retrospectively.Methods:Clinical data and CT images of 70 patients who performed flexible ureteroscopic lithotripsy because of single kidney stone in our center from January 2018 to December 2019 were retrospectively reviewed. There were 46 males and 24 females; their age was (47±12) years old. Stones were located on the left side in 28 cases and right side in 42 cases; 32 cases in the renal pelvis , 29 cases in the lower calyx, 6 cases in the middle calyx and 3 cases in the upper calyx. The free software ITK-SNAP 3.6.0 to segment kidney stones in 3D models with the CT image was used. The stone volume was calculated automatically after the segment. The largest section of the stone on the CT coronal plane was selected to measure the maximum length (D) and width (d) of the stone, the maximum diameter of the stone was D, and the stone area was calculated using the formula 0.25πDd. The patients were divided by the operation clearance into total clearance group and partial clearance group. The correlations between three stone burden parameters (volume, diameter, area) and operation time or clearance of the flexible ureteroscopic lithotripsy were compared. Simple linear regression model was also applied to compare three measurement methods. Then other factors which may affect the operation time was evaluated with the stepwise linear regression model, such as stone component and location.Results:The median operation time was 63(50, 84)min. Of 70 cases, 47 cases were in the stone-free group, with stone volume 633(248, 1 087)mm 3, maximum diameter 15(10, 19)mm, and area 82(49, 186)mm 2. 23 cases were in the non stone-free group, with volume 696(408, 1 418)mm 3, maximum diameter 15(12, 20)mm, area 105(73, 201)mm 2. There was no difference between the two groups in volume, maximum diameter and area of stones (all P>0.05). The stone-free rate of the diameter >2 cm group was 55% (6/11), ≤2 cm group was 70% (41/59). There was no significant difference between the two groups. Correlation between stone volume and operation time is the best. The correlation coefficient of stone volume is 0.58, of stone diameter is 0.33, of stone area is 0.34. Coefficients of determination of the stone volume linear regression is the best, too. R square of stone volume is 0.36, of stone diameter is 0.17, of stone area is 0.22. Forward stepwise regression model shows stone volume is the most important parameter which correlate with operation time. None of stone volume, diameter or area has significant correlation with the clearance of stone. Conclusion:Stone volume is the best predictive parameter of the stone burden because it has the best correlation with the operation time of the flexible ureteroscopic lithotripsy of the single kidney stone.

2.
Article in Chinese | WPRIM | ID: wpr-881386

ABSTRACT

@#Eleven alkaloids were isolated from the twigs and leaves of Ervatamia pandacaqui using chromatographic methods of silica gel, Sephadex LH-20, ODS, and HPLC.Their structures were elucidated by physical,chemical and spectroscopic methods and determined as voacristine 7-hydroxyindolenine (1),iboxygaine (2), 19S-hydroxyibogamine (3), 3-oxotabersonine (4), perivine (5), pericyclivine (6), rhazinalinol (7), geissoschizol (8), 3, 14-dihydroolivacine (9), vallesamine (10), and conolobine A (11), respectively.All compounds were isolated from this plant for the first time.

3.
Article in English | WPRIM | ID: wpr-880869

ABSTRACT

Gap junction (GJ) has been indicated to have an intimate correlation with adhesion junction. However, the direct interaction between them partially remains elusive. In the current study, we aimed to elucidate the role of N-cadherin, one of the core components in adhesion junction, in mediating connexin 43, one of the functional constituents in gap junction, via transforming growth factor-β1(TGF-β1) induction in osteoblasts. We first elucidated the expressions of N-cadherin induced by TGF-β1 and also confirmed the upregulation of Cx43, and the enhancement of functional gap junctional intercellular communication (GJIC) triggered by TGF-β1 in both primary osteoblasts and MC3T3 cell line. Colocalization analysis and Co-IP experimentation showed that N-cadherin interacts with Cx43 at the site of cell-cell contact. Knockdown of N-cadherin by siRNA interference decreased the Cx43 expression and abolished the promoting effect of TGF-β1 on Cx43. Functional GJICs in living primary osteoblasts and MC3T3 cell line were also reduced. TGF-β1-induced increase in N-cadherin and Cx43 was via Smad3 activation, whereas knockdown of Smad3 signaling by using siRNA decreased the expressions of both N-cadherin and Cx43. Overall, these data indicate the direct interactions between N-cadherin and Cx43, and reveal the intervention of adhesion junction in functional gap junction in living osteoblasts.


Subject(s)
Cadherins , Cell Communication , Connexin 43 , Osteoblasts , Transforming Growth Factor beta1
4.
Article in Chinese | WPRIM | ID: wpr-880481

ABSTRACT

In view of the inherent drawbacks of traditional medical equipment procurement mode, a decision-making model of medical equipment procurement based on Improved Markov model is proposed and the engineering evaluation practice is carried out. The data pool of medical equipment procurement of big data level is constructed, the core constraint factors of medical equipment procurement are perceived by deep learning, the decision model of medical equipment procurement under multi-dimensional constraints is constructed, and the Improved Markov model is introduced, and the observable decision-making scheme of medical equipment procurement is given. The results show that the model can give the decision-making scheme of medical equipment procurement with global optimal attribute under multi-dimensional constraints. It has obvious advantages in the balance of long-term demand and immediate demand, the optimization of procurement decision-making scheme, and the accuracy of patient demand prediction in a long period.

5.
Article in Chinese | WPRIM | ID: wpr-877549

ABSTRACT

OBJECTIVE@#To observe the effect of moxibustion on Nrf2/HO-1 signaling pathway in rats with diminished ovarian reserve (DOR), and to explore the protective mechanism of moxibustion on ovarian reserve function.@*METHODS@#Forty SD rats were randomly divided into a blank group, a model group, a moxibustion group and a hormone group, 10 rats in each group. The rats in the model group, moxibustion group and hormone group were treated with intragastric administration of tripterysium glycosides turbid liquid to prepare DOR model. The rats in the blank group were treated with intragastric administration of sodium chloride solution with the same volume, once a day for 14 days. The rats in the hormone group were treated with hormone sequential therapy for 14 days from the day of modeling; the rats in the moxibustion group were treated with moxibustion at bilateral "Shenshu" (BL 23) or "Guanyuan" (CV 4) and "Zhongwan" (CV 12) from the day of modeling, and the two groups acupoints were alternated every other day, 10 min each time, for 14 consecutive days. The estrus cycle was observed every day by vaginal exfoliated cell smear, and the estrus cycle disorder rate in each group was calculated. After the intervention, the HE staining was used to observe the histological morphology of ovaries; ELISA was used to detect the contents of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E@*RESULTS@#Compared with the blank group, the rate of estrus cycle disorder in the model group was increased (@*CONCLUSION@#Moxibustion could reduce the rate of estrus cycle disorder, improve the level of serum sex hormones and antioxidant stress in DOR rats, and the mechanism may be related to the regulation of Nrf2/HO-1 signaling pathway.


Subject(s)
Animals , Female , Humans , Moxibustion , NF-E2-Related Factor 2/metabolism , Ovarian Reserve , Rats , Rats, Sprague-Dawley , Signal Transduction
6.
Article in Chinese | WPRIM | ID: wpr-865804

ABSTRACT

Objective:To explore the application effect of four-track teaching mode (LBL: lecture-based learning; CBL: case-based learning; PBL: problem-based learning) in pediatric teaching of integrated traditional Chinese and Western Medicine.Methods:175 integrated traditional Chinese and Western medical students in seven-year program were divided into experimental group and control group. The experimental group was taught by four-track teaching mode, while students in the control group. Were applied with traditional teaching method ("LBL+CBL").Results:Students in the experimental group highly rated the four-track teaching mode and were better than those in the control group in the following aspects: satisfaction with teaching methods, interest in learning, the ability of creative thinking, the thinking ability of clinical diagnosis and treatment, and memorization ( P<0.05). And students' mid-term, final and comprehensive scores in the experimental group were significantly higher than those in the control group ( P<0.05). Conclusion:Compared with traditional teaching methods, the four-track teaching mode can improve the quality of pediatric teaching of integrated traditional Chinese and Western medical students, which should be further integrated and perfected in the teaching practice.

7.
Journal of Leukemia & Lymphoma ; (12): 684-687, 2020.
Article in Chinese | WPRIM | ID: wpr-862905

ABSTRACT

Objective:To investigate the effect of rapamycin in treatment of tyrosine kinase inhibitor (TKI)-resistant chronic myelogenous leukemia (CML) without ABL mutation.Methods:Flow cytometry was used to detect the positive expressions of p-mTOR and p-S6 in CD33 positive cells of 2 CML patients with TKI resistance in Jiangsu Province Hospital, and the influence of rapamycin on the positive expressions of p-mTOR and p-S6 in vitro.Results:Rapamycin in vitro inhibited the positive expressions of p-mTOR and p-S6 in CD33 positive cells. After 3 months of oral administration of rapamycin, the positive expressions of p-mTOR and p-S6 in CD33 positive cells were decreased, and the copy number of BCR-ABL fusion gene was also decreased simultaneously.Conclusion:Part of the resistance of CML patients to TKI may be related to the activation of intracellular signaling pathway of mTOR.

8.
Article in English | WPRIM | ID: wpr-828961

ABSTRACT

Homoeostasis depends on the close connection and intimate molecular exchange between extracellular, intracellular and intercellular networks. Intercellular communication is largely mediated by gap junctions (GJs), a type of specialized membrane contact composed of variable number of channels that enable direct communication between cells by allowing small molecules to pass directly into the cytoplasm of neighbouring cells. Although considerable evidence indicates that gap junctions contribute to the functions of many organs, such as the bone, intestine, kidney, heart, brain and nerve, less is known about their role in oral development and disease. In this review, the current progress in understanding the background of connexins and the functions of gap junctions in oral development and diseases is discussed. The homoeostasis of tooth and periodontal tissues, normal tooth and maxillofacial development, saliva secretion and the integrity of the oral mucosa depend on the proper function of gap junctions. Knowledge of this pattern of cell-cell communication is required for a better understanding of oral diseases. With the ever-increasing understanding of connexins in oral diseases, therapeutic strategies could be developed to target these membrane channels in various oral diseases and maxillofacial dysplasia.


Subject(s)
Bone and Bones , Cell Communication , Connexins , Metabolism , Physiology , Gap Junctions , Metabolism , Pathology , Homeostasis , Physiology , Humans , Mouth Diseases , Phosphorylation
9.
Article in English | WPRIM | ID: wpr-828960

ABSTRACT

Once pulp necrosis or apical periodontitis occurs on immature teeth, the weak root and open root apex are challenging to clinicians. Berberine (BBR) is a potential medicine for bone disorders, therefore, we proposed to apply BBR in root canals to enhance root repair in immature teeth. An in vivo model of immature teeth with apical periodontitis was established in rats, and root canals were filled with BBR, calcium hydroxide or sterilized saline for 3 weeks. The shape of the roots was analyzed by micro-computed tomography and histological staining. In vitro, BBR was introduced into stem cells from apical papilla (SCAPs). Osteogenic differentiation of stem cells from apical papilla was investigated by alkaline phosphatase activity, mineralization ability, and gene expression of osteogenic makers. The signaling pathway, which regulated the osteogenesis of SCAPs was evaluated by quantitative real time PCR, Western blot analysis, and immunofluorescence. In rats treated with BBR, more tissue was formed, with longer roots, thicker root walls, and smaller apex diameters. In addition, we found that BBR promoted SCAPs osteogenesis in a time-dependent and concentration-dependent manner. BBR induced the expression of β-catenin and enhanced β-catenin entering into the nucleus, to up-regulate more runt-related nuclear factor 2 downstream. BBR enhanced root repair in immature teeth with apical periodontitis by activating the canonical Wnt/β-catenin pathway in SCAPs.


Subject(s)
Animals , Berberine , Pharmacology , Cell Differentiation , Dental Papilla , Male , Osteogenesis , Periapical Periodontitis , Therapeutics , Rats , Stem Cells , Cell Biology , Metabolism , Wnt Signaling Pathway , Wnt3A Protein , Genetics , Metabolism , X-Ray Microtomography
10.
Article in Chinese | WPRIM | ID: wpr-873295

ABSTRACT

Objective::To investigate the effects of black bean juice with different stewing times on the appearance character and the content changes of effective components of Polygoni Multiflori Radix Praeparata. Method::HPLC was employed with Agilent ZORBAX Extend-C18 column (4.6 mm×250 mm, 5 μm), a gradient mobile phase of methanol (A)-water (B) was eluted (0-30 min, 5%-100%A; 30-40 min, 100%A), the flow rate was 1.0 mL·min-1, the injection volume was 10 μL and the column temperature was 35 ℃, detection wavelength was set at 280 nm. The contents of stilbene glycoside, emodin, emodin methyl ether, emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside in samples prepared at different processing times were simultaneously determined by HPLC. Result::The content of stilbene glycoside decreased gradually with the increase of stewing time, compared with 8 h, its content decreased by 76% at 64 h. The contents of emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside increased first, and then decreased, reaching the highest value at 24 h, and then decreased to the level similar to the content of 8 h after 40 h, and then fluctuated slightly. The contents of emodin and emodin methyl ether increased first, and then decreased, reached the maximum when stewed for 32 h, then decreased slowly and tended to be stable. Conclusion::The stewing time has significant influence on the content of various components in Polygoni Multiflori Radix Praeparata, and the changing trend is different, the processing time of Polygoni Multiflori Radix Praeparata shall be standardized. At the same time, it is not sufficient to take stilbene glycoside and anthraquinones as the indicator ingredients for this decoction pieces, the quality control indicators such as polysaccharides shall be considered.

11.
Article in Chinese | WPRIM | ID: wpr-872800

ABSTRACT

Objective::To establish a rapid evaluation method for Cinnamomi Cortex decoction pieces by near infrared spectroscopy. Method::The contents of coumarin, cinnamalol, cinnamic acid and cinnamaldehyde in 86 batches of Cinnamomi Cortex of different origins were determined by HPLC. And the NIR spectra of different batches of Cinnamomi Cortex were also collected. With NIR spectrum as independent variable and coumarin, cinnamalol, cinnamic acid and cinnamaldehyde as dependent variables, a quantitative analysis model of four components in cinnamon was established by partial least squares method. Result::The correlation coefficients (r) of coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde near infrared quantitative analysis models were 0.952 8, 0.977 7, 0.961 9, 0.992 2, root mean square error of cross(RMSEC) were 0.012 2, 0.006 1, 0.004 3, 0.82 g·g-1, root mean square errorof cross-validation(RMSECV) were 0.015 8, 0.011 2, 0.002 0, 1.481 1 g·g-1, and root mean square error of prediction(RMSEP) were 0.017 8, 0.010 3, 0.010 3, 0.005 5, 1.63 g·g-1. Conclusion::The established NIR quantitative analysis model of four active ingredients in Cinnamomi Cortex slices has a good accuracy, and provides a basis for rapid evaluation of the quality of Cinnamomi Cortex slices.

12.
Article in Chinese | WPRIM | ID: wpr-872743

ABSTRACT

Objective:To detect the changes of functional expression profile of energy metabolism related differential genes in sleep deprived rats before and after intervention by Tianwang Buxindan by microarray sequencing technology, so as to provide possible ideas and theoretical basis for the prevention and treatment of sleep deprivation. Method:The rats were randomLy divided into two groups: the Tianwang Buxindan group and the model group. The Tianwang Buxindan group was given the decoction of Tianwang Buxindan at the dose of 20 g·kg-1, and the model group was given the pure water of equal volume for 14 days. Taking liver, heart and hypothalamus as samples, high-throughput sequencing was used to obtain differential genes. Gene Ontology(Go)classification and kyoto Encyclopedia of Genes and Genomes (KEGG)pathway enrichment analysis were used to construct a co expression network with lncrna. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression levels of neuropeptide Y(NPY), bispecific phosphatase 1/mitogen-activated protein kinase phosphatase-1(DUSP1/MKP-1)and alpha-L-iduronidase(IDUA), three key genes with significant differences in energy metabolism. Result:The 321 differentially expressed genes were obtained, 231 of which were up-regulated and 90 down regulated, which mainly promoted the process of lipid metabolism, glucose metabolism and protein metabolism, participated in the synthesis and expression of fibrinogen, vitamin B6 and mesencephalic astrocyte-derived neurotrophic factor(MANF), and involved mitogen activated protein kinases(MAPK), p53 gene(p53), cyclic adenosine monophosphate(cAMP) and other signal pathways. Compared with the model group, the expression of IDUA significantly increased in the Tianwang Buxindan group (P<0.05), but decreased significantly in NPY and DUSP1(P<0.01). Conclusion:Tianwang Buxindan can interfere with the energy metabolism mechanism of sleep deprived rats in many ways. By down regulating the mRNA expression level of NPY and DUSP1 genes, it may activate the p38 MAPK signal pathway and affect the lipid metabolism.

13.
Article in Chinese | WPRIM | ID: wpr-872698

ABSTRACT

Objective:By studying the effects of Tianwang Buxindan on sleep quality, cognitive function, inflammatory factors and immune-related gene expression in sleep deprivation model rats, explore the effect of Tianwang Buxindan on the learning and memory process under sleep deprivation and its anti-inflammatory effects possible mechanism. Method:The 40 male SPF rats were used to simulate the sleep deprivation model by multi-platform water environment method, and were randomly divided into model group, Tianwang Buxindan group (20 g·kg-1) and estazolam group (0.1 mg·kg-1), set up a normal group, 10 in each group. A total of 4 weeks of sleep deprivation modeling was performed, and drug intervention was performed 2 weeks later. The model group and the blank group were given equal volumes of pure water. Electroencephalogram (EEG) evaluation of modeling and analysis of sleep structure and quality of rats, Morris water maze positioning navigation and space exploration experiment analysis of learning and memory ability of rats, application of enzyme-linked immunosorbent assay (ELISA) was used to detect the serum inflammatory factor interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), monocyte chemokine-1 (MCP-1) expression, Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA levels of EB virus inducible gene 3 (EBI3), extracellular signal-regulated kinase 5 (ERK5), and p21 activated protein kinase 4 (PAK4). Result:The sleep deprivation model was successfully built. Compared with blank group, the total sleep time, total duration of slow wave sleep and the duration of the first and second phases of slow wave sleep in the model group were significantly shortened (P<0.01). The incubation period on the upper platform, the total swimming distance and the time to reach the original platform for the first time increased significantly, while the number of times to cross the platform and the target quadrant significantly decreased (P<0.05,P<0.01). The expression levels of IL-1β,TNF-α and MCP-1 increased significantly (P<0.01), the mRNA expression levels of EBI3, ERK5 and PAK4 in the hypothalamus of the model group decreased significantly (P<0.01). Compared with the model group, the sleep quality of the rats in the Tianwang Buxindan group was significantly improved. The total sleep time, the total duration of slow wave sleep and the duration of the first phase of slow wave sleep were significantly increased (P<0.01). The incubation period on the platform, the total swimming distance and the time to reach the original platform for the first time are shortened, the number of times to cross the platform and the target quadrant time are extended (P<0.05,P<0.01), IL-1β, TNF-α, MCP-1 expression levels were significantly reduced (P<0.05,P<0.01), mRNA expression levels of EBI3, ERK5 and PAK4 in rat hypothalamus were significantly increased (P<0.05,P<0.01). Conclusion:Tianwang Buxindan can improve the sleep quality and learning and memory ability of sleep deprivation model rats, which may be related to the increase of the expression level of related inflammatory factors and its anti-inflammatory effect.

14.
Article in Chinese | WPRIM | ID: wpr-872647

ABSTRACT

Objective:To explore the effect of Anmeidan (AMD) on the learning and memory levels of sleep deprived rats through mitochondrial mediated hippocampal neuronal apoptosis pathway. Method:Forty-eight SD rats were randomly divided into blank group, model group, low, medium, high-dose AMD groups (4.86, 9.72, 19.44 g·kg-1·d-1) and estazolam group (0.1 mg·kg-1·d-1). Insomnia model was prepared by self-made sleep deprivation box for 14 days. Morris water maze was used to detect learning and memory levels, enzyme-linked immunosorbent assay (ELISA) was used to detect the expressions of cytochrome C (Cyt-C), cysteine aspartic acid protease-3 (Caspase-3) in hippocampus. Transmission electron microscopy (TEM) was used to observe the morphological structure of mitochondria in hippocampus. Protein and mRNA expressions of Cyt-C, Caspase-3, Bcl-2, Bax were detected by immunofluorescence (IF) and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) respectively. Result:In the model group, the incubation period of the platform and the total distance of swimming and the time of first arriving platform were prolonged, the number of platform crossing and the time of target quadrant movement were reduced, protein and mRNA expressions of Bcl-2 dropped, protein and mRNA expressions of Bax increased (P<0.01), and mitochondrial structure was abnormal with crista fracture, swelling and deformation. And protein and mRNA expressions of Cyt-C, Caspase-3 increased significantly (P<0.01). Low, medium and high-dose AMD groups could improve levels of space exploration and navigation of SD rats (P<0.01), increase protein and mRNA expressions of Bcl-2, decrease protein and mRNA expressions of Bax, improve the damage of mitochondria, and decrease the protein and mRNA expressions of Cyt-C, Caspase-3 (P<0.01). Conclusion:AMD can improve the learning and memory levels of SD rats, the effect is related to the mitochondrial mediated hippocampal neuronal apoptosis pathway and decrease of Cyt-C and Caspase-3 expressions.

15.
Acta Pharmaceutica Sinica ; (12): 294-297, 2020.
Article in Chinese | WPRIM | ID: wpr-789034

ABSTRACT

Four alkaloids were isolated from the total alkaloids of the twigs and leaves of Alstonia yunnanensis (Apocynaceae) by using silica gel, ODS, Sephadex LH-20, and HPLC chromatography. Structures were determined by physical, chemical and spectroscopic methods and identified as N4-methylpseudoakuammigine (1), pseudoakuammigine (2), vinorine (3), picraline (4). Among them, compound 1 is a new monoterpenoid indole alkaloid.

16.
Article in English | WPRIM | ID: wpr-827418

ABSTRACT

Wernekink commissure syndrome (WCS) is very rare. Four patients with WCS, admitted to our hospital from April to May 2018, were chosen for this study, and their clinical manifestations, imaging features, and etiology were retrospectively analyzed based on the literatures. All patients with WCS manifested as bilateral cerebellar ataxia such as symmetrical limb and trunk ataxia, but the degree of ataxia was asymmetrical distribution based on the anatomy. Dysarthria was the main and constant clinical manifestation of the syndrome. Ophthalmoplegia had great variability, and WCS with oculomotor nerve palsy may be considered as atypical WCS. The incidence of olivary degeneration and palatine myoclonus is relatively low, which may be related to the difference in the reported time intervals of cases. Changes in consciousness and emotion may be the characteristic of neglected WCS, which should be paid more attention. Cerebral infarction is the main etiology of WCS. We reported that cerebral infarction and WCS was the first symptom in a patient with systemic lupus erythematosus (SLE). We should pay more attention to special etiology in diagnosis and treatment of WCS.


Subject(s)
Cerebellar Ataxia , Cerebral Infarction , Humans , Lupus Erythematosus, Systemic , Retrospective Studies , Syndrome
17.
Chinese Journal of Biotechnology ; (12): 1590-1599, 2020.
Article in Chinese | WPRIM | ID: wpr-826818

ABSTRACT

β-defensin is a primary protein immune factor in channel catfish's (Ietalurus punetaus) resistance to pathogenic microorganisms. Its primary structure contains a signal peptide composed of 24 amino acid residues at the N-terminal and a mature peptide composed of 43 amino acid residues at the C-terminal. The mature peptide region is responsible for the biological activity of β-defensin. In the present study, a recombinant strain of Pichia pastoris that produces channel catfish β-defensin, was constructed to realize the biosynthesis of channel catfish β-defensin based on eukaryotic expression. First, the β-defensin gene "IPBD" was isolated from the skin of channel catfish by RT-PCR. After linking it with the expression vector pPICZA, pPICZA-IPBD was transferred into competent P. pastoris X-33 cells to obtain recombinant P. pastoris strains. The yeast transformants with multi-copy gene inserts were obtained by using the culture medium containing 1 000 μg/mL zeocin. Using BMM culture medium (without amino nitrogen culture medium) instead of BMMY culture medium (with amino nitrogen culture medium), the fermentation and culture conditions of the recombinant strain were optimized, and the optimal conditions for producing channel catfish β-defensin were determined as follows: the expression was induced for 96 h with 1.0% methanol at 28 °C , 250 r/min. Purified protein with molecular weight of 5.98 kDa was obtained by nickel affinity chromatography, and MALDI-TOF/TOF mass spectrometry proved that it was the expected recombinant IPBD. The antibacterial test results showed that the inhibitory rates of recombinant IPBD on Gram-positive Staphylococcus aureus and Listeria monocytogenes and Gram-negative Pseudomonas aeruginosa were 69.6%, 71.6% and 65.8%, respectively. This study provides a recombinant DNA technique for the development of small molecule natural antibacterial peptide from fish.

18.
Article in Chinese | WPRIM | ID: wpr-826726

ABSTRACT

OBJECTIVE@#To explore the therapeutic effect and partial mechanism of electroacupuncture (EA) for patients with insulin resistance (IR) polycystic ovary syndrome (PCOS).@*METHODS@#Seventy patients with IR-PCOS were randomly divided into an EA group (36 cases, 5 cases dropped off) and a medication group (34 cases, 4 cases dropped off). The patients in the medication group were treated with oral administration of metformin hydrochloride, 500 mg each time, twice a day. The patients in the EA group were treated with EA (continuous wave, 2 Hz of frequency) at Zusanli (ST 36), Zhongwan (CV 12), Qihai (CV 6), Yishu (EX-B 3), Shenshu (BL 23), Pishu (BL 20), Ciliao (BL 32) for 30 min, three times a week. One menstrual cycle or 4 weeks were taken as a course of treatment, and 3 continuous courses were given. The follow-up was 3 months. The lipid metabolism indexes of triacylglycerol (TG), total cholesterol (TC), high-density lipoprotein (HDL) and low-density lipoprotein (LDL), homeostasis model assessment-insulin resistance index (HOMA-IR) and testosterone (T) in serum were compared before and after treatment, and the clinical effects of the two groups were evaluated during the follow-up.@*RESULTS@#The total effective rate was 67.7% (21/31) in the EA group and 60.0% (18/30) in the medication group, with no significant difference between the two groups (>0.05). After treatment, the levels of serum T, HOMA-IR, LDL, TG and TC were decreased significantly in the two groups (<0.01, <0.05), and HDL was increased significantly (<0.01); the levels of TC in the EA group after treatment was lower than that in the medication group (<0.05).@*CONCLUSION@#EA may adjust some dyslipidemia in patients to correct IR and improve endocrine disorder of PCOS, which had superior/similar effects to metformin.


Subject(s)
Acupuncture Points , Electroacupuncture , Female , Humans , Insulin Resistance , Polycystic Ovary Syndrome , Therapeutics
19.
Article in Chinese | WPRIM | ID: wpr-827930

ABSTRACT

This research is to predict anti-Alzheimer's disease active constituents on the target of acetylcholinesterase(AChE) from Glycyrrhizae Radix et Rhizoma with the help of pharmacophore and molecular docking. AChE ligand-based pharmacophore model was set up and the molecular library of the constituents from Glycyrrhizae Radix et Rhizoma were established by collecting literature. Then the constituents from Glycyrrhizae Radix et Rhizoma were screen for the potential AChE inhibitory potency in silico through matching with the best pharmacophore model. The flexible docking was used to evaluate the interactions between compounds screened from pharmacophore model and AChE protein(PDB ID:4 EY7). The interactions were expressed including but not limited to CDOCKER interaction energy, hydrogen bonds and non-bonding interactions. The molecular library of Glycyrrhizae Radix et Rhizoma contains 44 chemical constituents. As for the pharmacophore model, six kinds of potential AChE inhibitory constituents from Glycyrrhizae Radix et Rhizoma were considered to be the promising compounds according to the results of searching 3 D database of pharmacophore model. The molecular docking was possessed and the interaction patterns were given to show the detail interactions. The compounds screening from the pharmacophore model were consistent with the existing studies to some degree, indicating that the virtual screen protocols of AChE inhibitory constituents from Glycyrrhizae Radix et Rhizoma based on pharmacophore and molecular docking was reliable.


Subject(s)
Drugs, Chinese Herbal , Glycyrrhiza , Molecular Docking Simulation , Rhizome , Triterpenes
20.
Article in Chinese | WPRIM | ID: wpr-871297

ABSTRACT

Adenosine, a purine nucleoside present in human cells, has a variety of biological functions and serves as an important immune regulator. It plays an important role in the differentiation, maturation and migration of immune cells, especially lymphocytes. Adenosine could bind to different receptors on the cell membrane to produce different effects. Extracellular adenosine level is a key factor in determining the subtypes of receptor. It is affected by synthesis, transport and metabolism, and closely related to the tissue energy state. As a key enzyme in adenosine metabolism pathway, adenosine deaminase has been proved to affect the immune function of adenosine by regulating extracellular adenosine level. In the past, researches on adenosine deaminase mainly focused on enzyme activity. In recent years, researchers have found other important functions of adenosine deaminase and classify it as moonlighting protein. Adenosine deaminase affects the development, proliferation, differentiation and functional activation of immune cells by binding to the anchoring proteins CD26 and adenosine receptors on the cell surface, exerting immune regulatory functions independently of its catalytic functions. In recent years, the immunomodulatory functions of adenosine deaminase have attracted increasing attention. This paper reviewed the new progress in the regulatory functions of adenosine deaminase in the immune system.

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