The high content of sucrose and raffinose reduces the prebiotic value of soybean oligosaccharides. Fructan sucrases can catalyze the conversion of sucrose and raffinose to high-value products such as fructooligosaccharides and melibiose. To obtain a fructan sucrase that can efficiently convert soybean oligosaccharides, we first mined the fructan sucrase gene from microorganisms in the coastal areas of Xisha Islands and Bohai Bay and then characterized the enzymatic and catalytic properties of the enzyme. Finally, recombinant extracellular expression of this gene was carried out in Bacillus subtilis. The results showed that a novel fructan sucrase, BhLS 39, was mined from Bacillus halotolerans. With sucrose and raffinose as substrates, BhLS 39 showed the optimal temperatures of 50 ℃ and 55 ℃, optimal pH 5.5 for both, and K_cat/K_m ratio of 3.4 and 6.6 L/(mmol·s), respectively. When 400 g/L raffinose was used as the substrate, the melibiose conversion rate was 84.6% after 30 min treatment with 5 U BhLS 39. Furthermore, BhLS 39 catalyzed the conversion of sucrose to produce levan-type-fructooligosaccharide and levan. Then, the recombinant extracellular expression of BhLS 39 in B. subtilis was achieved. The co-expression of the intracellular chaperone DnaK and the extracellular chaperone PrsA increased the extracellular activity of the recombinant BhLS 39 by 5.2 folds to 17 U/mL compared with that of the control strain. BhLS 39 obtained in this study is conducive to improving the quality and economic benefits of soybean oligosaccharides. At the same time, the strategy used here to enhance the extracellular expression of BhLS 39 will also promote the efficient recombinant expression of other proteins in B. subtilis.
Oligosaccharides/metabolism* ; Glycine max/metabolism* ; Bacillus subtilis/metabolism* ; Sucrase/biosynthesis* ; Raffinose/metabolism* ; Fructans/metabolism* ; Sucrose/metabolism* ; Bacillus/genetics* ; Recombinant Proteins/biosynthesis* ; Bacterial Proteins/biosynthesis*

Objective:To summarize the evidence of pelvic floor muscle training for the prevention and treatment of postpartum urinary incontinence, providing guidance and reference for clinical practice.Methods:According to the "6S" pyramid model, clinical decision-making, guidelines, and systematic reviews on pelvic floor muscle training for the prevention and treatment of postpartum urinary incontinence were searched in UpToDate, British Medical Journal (BMJ) Best Practice, National Institute for Health and Care Excellence, Scottish Intercollegiate Guideline Network, New Zealand Guideline Group, Guidelines International Network, Medlive, Joanna Briggs Institute (JBI) Evidence-Based Health Care Center Database, Cochrane Library, professional association website, Embase, PubMed, Web of Science, China National Knowledge Infrastructure, China Biology Medicine disc, WanFang Data, and VIP. The search period was from March 2013 to March 2023. Two trained researchers evaluated the quality of literature and integrated and extracted evidence.Results:A total of 22 articles were included, including 8 clinical decision-making, 6 guidelines, 7 systematic reviews, and 1 expert consensus. Twenty-one best pieces of evidence were summarized from 5 aspects, consisting of risk factors, prevention, evaluation, treatment and health guidance for postpartum urinary incontinence.Conclusions:The best evidence for the prevention and treatment of postpartum urinary incontinence through pelvic floor muscle training summarized is convenient for medical and nursing staff to conduct scientific urinary incontinence assessment, pelvic floor muscle training education and guidance for pregnant and postpartum women.

ObjectiveBased on serum pharmacochemistry and ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） the transitional components in the serum of rats after intragastric administration of water extract of Alismatis Rhizoma（AR）and salt-processed Alismatis Rhizoma（SAR） were compared. MethodSD rats were randomly divided into blank group, AR group（10 g·kg^-1） and SAR group（10 g·kg^-1）, 3 rats in each group, the administration groups were given AR and SAR aqueous extracts by gavage, respectively, and the blank group was given an equal volume of drinking water by gavage once in the morning and once in the evening, for 3 consecutive days. Sixty minutes after the last administration, blood was collected from the eye orbits, and the serum samples were prepared. The serum samples were prepared on an ACQUITY UPLC BEH C18 column（2.1 mm×50 mm, 1.7 μm） with the mobile phase of acetonitrile（A）-0.1% formic acid aqueous solution（B） in a gradient elution（0-10 min, 10%-50% A; 10-27 min, 50%-95%A; 27-27.1 min, 95%-10% A; 27.1-30 min, 10%A）, the data were collected at a flow rate of 0.3 mL·min^-1 in positive ion mode with a scanning range of m/z 100-1 200. Based on the self-constructed chemical composition library of AR, the total ion flow diagrams and secondary MS fragmentation information of the aqueous extracts of AR and SAR, as well as the administered serum and the blank serum, were compared with each other by UNIFI 1.9.2, so as to deduce the possible blood-migrating constituents and their cleavage patterns in the aqueous extracts, and the response intensity ratios of each chemical component were calculated before and after processing. ResultA total of 20 components, including 5 prototypical components and 15 metabolites, were analyzed and deduced from the serum of rats given aqueous extract of AR. And 14 components, including 5 prototypical components and 9 metabolites, were analyzed and deduced from the serum of rats given aqueous extract of SAR. Of these, 13 components were common to both of them, including 5 prototypical components and 8 metabolites. The 5 prototypical components were 16-oxoalisol A, alisol A 24-acetate, alisol A, alisol B and alisol C. The metabolites were mainly involved in phase Ⅰ metabolism（oxidation） and phase Ⅱ metabolism（glucuronidation）. There was a big change in the intensity of response of the common components before and after salt-processing, and the response intensities of the prototypical components, 16-oxoalisol A, alisol B and alisol C, were elevated, while the type and response intensity of metabolites were generally decreased, and it was hypothesized that the metabolic rate of terpenoids might be slowed down after salt-processing of AR, so that the blood-migrating constituents could participate in the metabolism of the body more in the form of prototypes. ConclusionSalt-processing of AR may promote the absorption of prototypical components into the blood by slowing down the metabolic rate of terpenoids, which can provide support for the research on material basis of AR and SAR.

ObjectiveTo investigate the effect of microemulsion on the distribution of index components in different phases of Zexietang extract based on high performance liquid chromatography（HPLC） and phase separation process. MethodParticle size meter and transmission electron microscope were used to characterize the colloidal particles in blank microemulsion， aqueous extract of Zexietang and microemulsion extract of Zexietang. The phase separation process was established by high-speed centrifugation and dialysis， and based on this process， the aqueous extract and microemulsion extract of Zexietang were separated into the true solution phase， the colloidal phase and the precipitation phase， respectively. The contents of six components， including atractylenolide Ⅲ， atractylenolide Ⅱ， 23-acetyl alisol C， alisol A， alisol B and alisol B 23-acetate， were determined by HPLC with the mobile phase of water（A）-acetonitrile（B） for gradient elution（0-5 min， 40%-43%B； 5-20 min， 43%-45%B； 20-45 min. 45%-60%B； 45-75 min， 60%-80%B）. The solubility of the index components in water and microemulsion was determined by saturation solubility method. ResultThe colloidal particles in the aqueous extract， microemulsion extract and blank microemulsion were all spherical， and the particle size， polydispersity index（PDI） and Zeta potential of the colloidal particles were in the order of aqueous extract >microemulsion extract >blank microemulsion. The results of phase separation showed that the colloidal phase and the true solution phase could be completely separated by dialysis for 2.5 h， and the phase separation process was tested to be stable and feasible. Compared with the aqueous extract of Zexietang， the use of microemulsion as an extraction solvent could increase the contents of atractylenolide Ⅲ， 23-acetyl alisol C， atractylenolide Ⅱ ， alisol A， alisol B and alisol B 23-acetate by 3.75， 6.82， 35.47， 10.66， 35.41， 27.75-fold， and could increase the extraction efficiencies of the latter five constituents by 2.03， 1.15， 1.70， 6.43， 5.53 times. The solubility test showed that the microemulsion could significantly improve the solubility of atractylenolide Ⅱ， alisol A， alisol B and alisol B 23-acetate， but it had less effect on the solubility of atractylenolide Ⅲ and 23-acetyl alisol C. ConclusionMicroemulsion can improve the extraction efficiency and increase the distribution of the index components in the colloidal phase state of Zexietang to different degrees， providing a reference for the feasibility of microemulsion as an extraction solvent for traditional Chinese medicine.

ObjectiveTo evaluate the pharmacological effect of Alismatis Rhizoma （AR） and its processed product on rats with edema of kidney Yin deficiency and explore the mechanism. MethodA total of 42 male SPF SD rats were randomized into normal group （equivalent volume of distilled water）， model group （equivalent volume of distilled water）， positive medicine Liuwei Diguangwan group （1.4 g·kg^-1）， low- and high-dose AR groups （1， 4 g·kg^-1， respectively）， and low- and high-dose salt-processed AR （SAR） groups （1， 4 g·kg^-1， respectively）， with six rats in each group. Adriamycin （tail vein injection） and thyroxine （gavage） were used to induce edema of kidney Yin deficiency in rats except the normal group. The administration lasted 4 weeks for all the groups. After the last administration， histopathological changes of rat kidneys were observed based on hematoxylin-eosin （HE） staining. Serum content of triiodothyronine （T₃）， thyroxine （T₄）， follicle-stimulating hormone （FSH）， and testosterone （T） was determined by radioimmunoassay， and serum content of creatinine （CREA）， urea （UREA），cholesterol （CHOL） and triglyceride （TG） by automatic biochemical analyser. The levels of gonadotropin-releasing hormone （GnRH）， cyclic adenosine monophosphate （cAMP）， and cyclic guanosine monophosphate （cGMP） in plasma were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of aquaporin（AQP）-1 and AQP-2 and the transcription of mRNA in kidney were measured by immunohistochemistry and real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， respectively. ResultCompared with normal group， the rats in model group showed decrease in body mass and urine volume （P<0.01）， increase in water consumption （P<0.05）， infiltration of a large number of inflammatory cells and fibrous tissue proliferation in the kidney， rise of the expression and transcript levels of T₃， T₄， cAMP/cGMP， CREA， FSH， AQP-1， and AQP-2 （P<0.01）， the contents of CHOL and TG were significantly increased （P<0.05）， and reduction in the levels of GnRH and T （P<0.01）. Body mass increased in both the low- and high- dose groups of AR and SAR compared with that in model group， with significant differences between the low-dose AR group and the low-dose SAR group （P<0.01）. Moreover， compared with model group， low- and high-dose AR and SAR insignificantly increased the urine volume of rats， reduced the inflammatory cells in kidney tissues， significantly decreased the levels of T₄， cAMP/cGMP， UREA， CREA， FSH， CHOL and TG in serum （P<0.05，P<0.01）， and elevated the level of GnRH （P<0.01）， high-dose AR， low- and high-dose SAR significantly lowered the transcription levels of AQP-1 and AQP-2 mRNA in the kidneys of rats （P<0.01）. ConclusionBoth AR and SAR alleviated the edema of kidney Yin deficiency in rats by down-regulating the expression of AQP-1 and AQP-2 and correcting the hypothalamic-pituitary-gonadal （HPG） axis disorder.

Single-cell RNA sequencing (scRNA-seq) is generally used for profiling transcriptome of individual cells.The droplet-based 10X Genomics Chromium (10X) approach and the plate-based Smart-seq2 full-length method are two frequently used scRNA-seq platforms,yet there are only a few thorough and systematic comparisons of their advantages and limitations.Here,by directly comparing the scRNA-seq data generated by these two platforms from the same samples of CD45-cells,we systematically evaluated their features using a wide spectrum of analyses.Smart-seq2 detected more genes in a cell,especially low abundance transcripts as well as alterna-tively spliced transcripts,but captured higher proportion of mitochondrial genes.The composite of Smart-seq2 data also resembled bulk RNA-seq data more.For 10X-based data,we observed higher noise for mRNAs with low expression levels.Approximately 10％-30％ of all detected tran-scripts by both platforms were from non-coding genes,with long non-coding RNAs (lncRNAs)accounting for a higher proportion in 10X.10X-based data displayed more severe dropout prob-lem,especially for genes with lower expression levels.However,10X-data can detect rare cell types given its ability to cover a large number of cells.In addition,each platform detected distinct groups of differentially expressed genes between cell clusters,indicating the different characteristics of these technologies.Our study promotes better understanding of these two platforms and offers the basis for an informed choice of these widely used technologies.

Objective To explore the effect of adipose-derived stem cells (ASCs) on the skin expansion rate in rabbit.Methods The rabbit ASCs were isolated from fat tissue and cultured in vitro.The ADSCs were identified by cell immunofluorescence and marked by Edu staining.20 new Zealand rabbits were randomly divided into experimental(n =10) and control group(n =10).An area of 1.5 cm ×1.5 cm on the one side back of each rabbit was tattooed and one 30 ml round expander was implanted subcutaneously.ASCs suspension (1 ml) was injected subcutaneously in the experimental group,while serum free DMEM medium(1 ml) in control group.The expansion was proceeded regularly under constant pressure for 4 weeks.The expanded tattooed square area was measured on the 7th,14th,28th day and analyzed statistically.The expanded skin was harvested for histological study.Immunohistochemical staining was used to detect the expression of vascular endothelial cell marker CD31,and the microvessel density determination.The expression of epidermal growth factor (EGF) and vascular endothelial growth factor(VEGF)was detected by ELISA for skin tissue specificity.Western Blot was used for detection of CK19 in the epidermal cells.Results The expanded skin thickness and expansion rate in experimental group were significant higher than those in control group (P ＜ 0.05).Compared with control group,the expression of CK19,CD31 and EGF,VEGF,as well as the microvessel density were all markedly increased in experimental group(P ＜0.05).Conclusions ASCs can increase the expansion rate of skin tissue by promotion of angiogenesis and tissue regeneration.

Objective To explore the effect of adipose-derived stem cells (ASCs) on the skin expansion rate in rabbit.Methods The rabbit ASCs were isolated from fat tissue and cultured in vitro.The ADSCs were identified by cell immunofluorescence and marked by Edu staining.20 new Zealand rabbits were randomly divided into experimental(n =10) and control group(n =10).An area of 1.5 cm ×1.5 cm on the one side back of each rabbit was tattooed and one 30 ml round expander was implanted subcutaneously.ASCs suspension (1 ml) was injected subcutaneously in the experimental group,while serum free DMEM medium(1 ml) in control group.The expansion was proceeded regularly under constant pressure for 4 weeks.The expanded tattooed square area was measured on the 7th,14th,28th day and analyzed statistically.The expanded skin was harvested for histological study.Immunohistochemical staining was used to detect the expression of vascular endothelial cell marker CD31,and the microvessel density determination.The expression of epidermal growth factor (EGF) and vascular endothelial growth factor(VEGF)was detected by ELISA for skin tissue specificity.Western Blot was used for detection of CK19 in the epidermal cells.Results The expanded skin thickness and expansion rate in experimental group were significant higher than those in control group (P ＜ 0.05).Compared with control group,the expression of CK19,CD31 and EGF,VEGF,as well as the microvessel density were all markedly increased in experimental group(P ＜0.05).Conclusions ASCs can increase the expansion rate of skin tissue by promotion of angiogenesis and tissue regeneration.

Objective To investigate the effects of local injection of rabbit adipose-derived mesenchymal stem cells (ADSCs) on the healing of skin deep partial-thickness scald wound of rabbit.Methods ADSCs were isolated from adipose tissue of one New Zealand rabbit and then sub-cultured.ADSCs of the third passage were used in the following experiments.Twenty-four rabbits were divided into ADSCs group (n =12) and control group (n =12) according to the random number table,and one deep partial-thickness scald wound with diameter of 5 cm on the two sides of the back near the buttocks was made.From post injury day (PID) 2,2 mL suspension of EdU-labeled ADSCs with the number of 5 × 105 per mL was subcutaneously injected in wounds of rabbits in ADSCs group,while the rabbits in control group were given 2 mL serum-free DMEM until the wounds were healed.Wound healing processes of rabbits in two groups were observed every day,and the healing time was recorded.On PID 7,14,21,and 28,areas of wound of three rabbits in two groups were measured and the healing rates were calculated,respectively.The healed wound tissue was harvested to observe the morphology by HE staining,and the expression of collagen fiber was observed by Masson staining.The distribution of EdU-labeled ADSCs in healed wound tissue on PID 28 was observed by inverted fluorescence microscope.The expressions of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) of healed wound tissue on PID 7,14,and 21 were detected by enzymelinked immunosorbent assay.Data were processed with analysis of variance of factorial design and paired samples t test.Results (1) The wound healing time of rabbits in ADSCs group was (19.5 ± 1.1) d post injury,which was significantly shorter than that in control group [(23.3 ± 1.5) d,t =4.50,P ＜ 0.05].On PID 7,wounds of rabbits in two groups were dry with no obvious exudation,and redness and swelling around wounds disappeared gradually,the wound healing rate of rabbits in ADSCs group was (15.1 ± 2.4)％,which was close to that in control group [(13.7±3.1)％,t =1.20,P ＞0.05].On PID 14,wounds of rabbits in ADSCs group were dry and scabbed obviously,and the wound healing rate was (73.1 ± 5.7) ％,while wounds of rabbits in control group were little scabbed with little exudation,and the wound healing rate was significantly lower than that in ADSCs group [(52.9 ± 5.1)％,t =8.06,P ＜ 0.01].On PID 21,wounds of rabbits in ADSCs group were generally healed,and the wound healing rate was (95.6 ± 3.0) ％,while a few wounds still existed in rabbits of control group,and the wound healing rate was significantly lower than that inADSCs group [(78.6±3.7)％,t =9.73,P ＜0.01].On PID 28,wounds of rabbits in two groups were totally healed with the healing rate of 100％,and texture and microvascular responses of healed wound tissue in ADSCs group were better than those in control group.(2) On PID 7,fibroblasts in healed wound tissue of rabbits in two groups were all increased,and there were little vascular and collagen fiber proliferation with no obvious differences.On PID 14,the number of fibroblasts in healed wound tissue of rabbits in ADSCs group was more than that in control group,and the collagen fibers in healed wound tissue of rabbits in ADSCs group were arranged in dense and uniform,while those in control group were sparse and irregular.On PID 21,skin layers were differentiated in healed wound tissue of rabbits in two groups,and collagen fibers in healed wound tissue of rabbits in ADSCs group were still denser than that in control group.On PID 28,newborn skin was well differentiated in healed wound tissue of rabbits in ADSCs group,which was better than that in control group.There were a lot of thick collagen fibers in healed wound tissue of rabbits in two groups,and EdU-labeled ADSCs were involved in skin texture of rabbits in ADSCs group.(3) The expressions of VEGF and EGF in healed wound tissue of rabbits in two groups were similar on PID 7 (with t values respectively 0.70 and 0.91,P values above 0.05),which in ADSCs group were significantly higher than those in control group on PID 14 and 21 (with t values from 2.85 to 4.81,P values below 0.01).Conclusions The transplantation of ADSCs can promote the wound healing of skin deep partial-thickness scald wound of rabbit and shorten the wound healing time.

There is an increasing interest and popularity of Chinese herbal medicine worldwide, which is accompanied by increasing concerns about its effectiveness and potential toxicity. Several ingredients, such as polyphenolic compounds berberine, flavonoids, and curcumin, have been studied extensively by using various animal models. Effectiveness of treatment and amelioration of metabolic syndromes, including insulin resistance and dyslipidemia, has been demonstrated. This review summarizes the major checkpoints and contributing factors in regulation of exogenous and endogenous lipid metabolism, with particular emphasis centered on triglyceride-rich and cholesterol-rich lipoproteins. Available experimental evidence demonstrating the lipid-lowering effect of berberine, flavonoids and curcumin in cell culture and animal models is compiled, and the strengths and shortcomings of experimental designs in these studies are discussed.
Animals ; Drugs, Chinese Herbal ; therapeutic use ; Dyslipidemias ; drug therapy ; Humans ; Lipid Metabolism ; Lipoproteins ; metabolism ; Metabolic Syndrome ; drug therapy ; Polyphenols ; therapeutic use