Aim To produce poly (lactic-co-glycolic acid) (PLGA) microspheres, containing a staphylokinase variant (K35R, DGR) with reduced immunogenecity and antiplatclet aggregation activities,which allowed the preservation of protein stability during both particle processing and drug release.Methods DGR-loaded microspheres were fabricated using a double emulsion-solvent evaporation technique. The effects of preparative parameters, such as stirring rate, polymer concentration, and the excipients of both internal and external aqueous phase (W2 ), on DGR encapsulation efficiency and microsphere characteristics were investigated. In vitro and in vivo release of DGR were conducted and the cause for instability of DGR during release was also investigated. Results Moderate ultrasonic treatment of aqueous DGR/dichloromethane mixtures caused approximately. Eighty four per cent DGR denaturation.However, the activity recovery of DGR almost amounted to 100% when 2% polyvinyl alcohol (PVA) was added into the aqueous phase. It was found that NaCl in the external water phase significantly increased DGR encapsulation efficiency. Furthermore, NaCl in the external water phase played a role in determining size and surface morphology of microsphere. In vitro release test showed a burst release of DGR from microspheres, followed by sustained release of 50% total activity over 15 days. In vivo experiments showed that DGR released from microspheres sustained 5 days. Denaturation of DGR within microspheres might be resulted from acidic microclimate. Conclusion The stability of DGR was effectively protected during microencapsulation and a relatively high encapsulation efficiency of DGR was obtained. PLGA microspheres could be an effective carrier for DGR.

Objective: To investigate the clinical efficacy and safety of Shure granule combined with estradiol valerate in the prevention of intrauterine adhesions in induced abortion.Methods: Selected 240 cases of artificial abortion were divided into observation group and control group according to the order of admission, 120 cases each.Patients in the control group were treated with estradiol valerate.The observation group was treated with Shure granules on the basis of the control group;The treatment time was 3 weeks.The time of vaginal bleeding and the amount of bleeding, menstruation, retidal and adverse drug reactions of the two groups were observed;The levels of serum fibronectin (FN), human laminin (LN) and vascular endothelial growth factor (VEGF) were compared between the two groups before and after the operation.The adhesion of the 12 cases in the two cases was compared.Results: The time of vaginal bleeding and the amount of bleeding in the observation group were significantly better than those in the control group (P＜0.05), and the normal rate of menstrual volume was higher than that of the control group (P＜0.05).The serum levels of FN, LN and VEGF in the observation group were significantly lower than those in the control group 2 weeks after the operation (P＜0.05).After 12 months, the incidence of intrauterine adhesions in the observation group was significantly lower than that in the control group (P＜0.05).There was no significant difference in the incidence of adverse drug reactions between the two groups (P＞0.05).Conclusion: The efficacy of Shure granule combined with estradiol valerate in preventing intrauterine adhesions in induced abortion is superior to estradiol valerate alone.

OBJECTIVETo investigate the effect of environmental risk factors on the development of stroke.

METHODSWith the use of cold-stimuli plus high-salt intake as environmental risk factors, a hypertension model with the complication of stroke was established in rats, then, a new technique, suppression subtractive hybridization (SSH), was used to identify the differential genes which specifically expressed in total cerebrum tissue of rat in each group. Comparison was made between control group and stroke group.

RESULTSBy the application of SSH, a total of 576 clones were generated in this study from two subtractive libraries, among them 456 clones were usable and were analyzed. Genes for cell/organs defense were down-regulated in stroke group and metabolism transcripts were shown to be up-regulated (P<0.01).

CONCLUSIONCell/organs defense genes may play important roles in the development of stroke. The above findings suggested that environmental risk factors could genetically alter individual sensitivity to stroke.

Animals ; Brain ; metabolism ; pathology ; Cell Division ; genetics ; Cold Temperature ; DNA, Complementary ; chemistry ; genetics ; Expressed Sequence Tags ; Gene Expression Regulation ; drug effects ; Immunity, Innate ; genetics ; Male ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Rats ; Rats, Wistar ; Sequence Analysis, DNA ; Signal Transduction ; genetics ; Sodium Chloride, Dietary ; administration & dosage ; Stroke ; genetics

DNA barcoding has been widely used for herb identification in recent decades,enabling safety and innovation in the field of herbal medicine.In this article,we summarize recent progress in DNA bar-coding for herbal medicine to provide ideas for the further development and application of this tech-nology.Most importantly,the standard DNA barcode has been extended in two ways.First,while conventional DNA barcodes have been widely promoted for their versatility in the identification of fresh or well-preserved samples,super-barcodes based on plastid genomes have rapidly developed and have shown advantages in species identification at low taxonomic levels.Second,mini-barcodes are attractive because they perform better in cases of degraded DNA from herbal materials.In addition,some mo-lecular techniques,such as high-throughput sequencing and isothermal amplification,are combined with DNA barcodes for species identification,which has expanded the applications of herb identification based on DNA barcoding and brought about the post-DNA-barcoding era.Furthermore,standard and high-species coverage DNA barcode reference libraries have been constructed to provide reference se-quences for species identification,which increases the accuracy and credibility of species discrimination based on DNA barcodes.In summary,DNA barcoding should play a key role in the quality control of traditional herbal medicine and in the international herb trade.

Objective:To explore the effect of intervention model based on comprehensive nutrition management on glucose and lipid metabolism and pregnancy outcome in patients with gestational diabetes mellitus (GDM).Methods:104 GDM patients admitted to Shanxi Bethune Hospital from February 2022 to March 2023 were randomly divided into control group and experimental group, with 52 cases in each group. The control group implemented routine management measures and nutrition guidance, while the experimental group implemented an intervention model based on comprehensive nutrition management on the basis of routine management. The indexes of glucose and lipid metabolism (glycosylated hemoglobin, fasting blood glucose, 2 h postprandial blood glucose, total cholesterol, triglyceride and low density lipoprotein cholesterol), pregnancy outcome, self-management ability and self-efficacy were compared between the two groups before and after intervention.Results:Before the intervention, there was no significant difference in general situation, glucose and lipid metabolism index, self-management ability and self-efficacy between the two groups ( P>0.05). After the intervention, the level of glucose and lipid metabolism index in the experimental group was significantly lower than that in the control group, with statistical significance ( P<0.05). The incidence of adverse pregnancy outcome in the experimental group was significantly lower than that in the control group, with statistical significance ( P<0.05). The scores of self-management ability and self-efficacy in the experimental group were significantly higher than those in the control group, with statistical significance ( P<0.05). Conclusion:The intervention model based on comprehensive nutrition management can effectively improve the glucose and lipid metabolism index and pregnancy outcome of GDM patients, and significantly improve their self-management ability and self-efficacy related to nutrition management, which has high clinical application and promotion value.

Allergic rhinitis (AR) is an allergic disease of nasal mucosa, currently the consensus has been reached that there is a Th1/Th2 imbalance in AR′s pathogenesis, meanwhile there are defects in the number and/or function of Treg cells. Mesenchymal stem cells (MSCs) have characteristics of multiple differentiation potential, low immunogenicity and immunomodulation function, which can be migrated and implanted into the nasal inflammatory mucosal, recovering the Th1/Th2 immune balance and up-regulate the Treg cells by immunomodulatory function to improve AR. For the present, the treatment of AR by MSCs is basically at the animal experiment stage. This paper reviews the progress of MSCs in AR′s treatment and provides a basis for the clinical treatment of MSCs.

ObjectiveTo investigate the different effects of Yunvjian with or without Achyranthis Bidentatae Radix on glucose and lipid metabolism and inflammatory response in diabetic rats with the syndrome of Yin deficiency and internal heat. MethodThe rat model of diabetes due to Yin deficiency and internal heat was established by feeding with a high-sugar and high-fat diet and injection of thyroxine and streptozotocin. The successfully modeled rats were randomized into model control， Yunvjian without Achyranthis Bidentatae Radix （11.8 g·kg^-1）， Yunvjian with Achyranthis Bidentatae Radix （12.8 g·kg^-1）， and Achyranthis Bidentatae Radix （1.0 g·kg^-1） groups （n=10）， and another 10 rats were taken as the normal control group. Each group was administrated with corresponding drugs or saline by gavage for 28 days. The fasting blood glucose （FBG）， fasting insulin （FINS）， total cholesterol （TC）， triglycerides （TG）， low-density lipoprotein cholesterol （LDL-C）， and high-density lipoprotein cholesterol （HDL-C） in rats were measured. Enzyme-linked immunosorbent assay was employed to determine the levels of cyclic adenosine monophosphate （cAMP）， cyclic guanosine monophosphate （cGMP）， triiodothyronine （T₃）， thyroxine （T₄）， interleukin （IL）-1β， IL-6， tumor necrosis factor （TNF）-α， and C-reactive protein （CRP） in the serum. The histopathological changes of the liver were observed. The expression of lipoxygenase-2 （COX-2） was detected by immunofluorescence. The mRNA levels of nuclear transcription factors-κB （NF-κB）， monocyte chemotactic protein-1 （MCP-1）， and intercellular adhesion molecule-1 （ICAM-1） were determined by real-time polymerase chain reaction （Real-time PCR）.Western blot was employed to determine the protein levels of NF-κB in hibitory protein（IκB） kinase β （IKKβ）， IκBα， and phosphorylated IκBα （p-IκBα） in the liver and the protein levels of NF-κB in the cytoplasm and nucleus. ResultCompared with the normal group， the model group showed elevated levels of FBG， FINS， insulin resistance index， TC， TG， LDL-C， cAMP， T₃， T₄， IL-1β， IL-6， TNF-α， and CRP， up-regulated mRNA levels of NF-κB， MCP-1， and ICAM-1， and up-regulated protein levels of COX-2， p-IκBα， and nuclear NF-κB （P<0.01）. Compared with the model group， Yunvjian without Achyranthis Bidentatae Radix lowered the levels of FBG， FINS， insulin resistance index， TC， TG， LDL-C， cAMP， T₃， T₄， IL-1β， IL-6， TNF-α， and CRP， down-regulated the mRNA levels of NF-κB， MCP-1， and ICAM-1， and down-regulated the protein levels of COX-2， p-IκBα and nuclear NF-κB （P<0.05， P<0.01）. Compared with the Yunvjian without Achyranthis Bidentatae Radix， Yunvjian with Achyranthis Bidentatae Radix showed lowered levels of FBG， FINS， insulin resistance index， and inflammatory cytokines， down-regulated mRNA levels of NF-κB， MCP-1， and ICAM-1， and down-regulated protein levels of p-IκBα and nuclear NF-κB （P<0.05， P<0.01）. ConclusionAchyranthis Bidentatae Radix can enhance the performance of Yunvjian in reducing blood glucose and inhibiting inflammation in diabetic rats with the syndrome of yin deficiency and internal heat by down-regulating the IKK/IκB/NF-κB signaling pathway.

Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.

Objective: To investigate the correlation between the proliferation inhibition effect of glucocorticoid (GC) on peripheral blood mononuclear cell (PBMC) and the pure tone average (PTA) improvement in SSNHL patients. Methods: Sixty inpatients with SSNHL were included from July 2013 to October 2015 in Nanjing Drum Tower Hospital, Medical School of Nanjing University. Peripheral venous blood was collected before receiving treatment, then the PBMC was isolated for GC proliferation inhibition. PBMCs of each patient were cultivated into 4 groups: Group A: PBMCs+ Medium; Group B: PBMCs+ Medium+ lipopolysaccharide (LPS, 1 μmol/L); Group C: PBMCs+ Medium+ LPS+ Dexamethasone; Group D: Medium. PBMCs were maintained in a humidified 5% CO₂ atmosphere at 37°C and were observed after 24 hours. 5-diphenyltetrazolium bromide (MTT) was used to measure PBMC proliferation inhibition rate. The PBMC proliferation inhibition rates were calculated according to the absorbance at 490 nm wavelength under a microtiter plate reader. Independent sample t tests of PBMC proliferation inhibition rate were performed between different groups. χ² tests were performed between gender, affected ear side, accompanied by vertigo or not, audiometric curve, time period from onset to treatment, PBMC proliferation inhibition rate and the improvement of pure tone average (PTA). Linear correlation analyses were performed between PBMC proliferation inhibition rate, the time period from onset to treatment and the hearing improvement. Results: The proliferation inhibition effect of GC on PBMC varied significantly among patients. The PBMC proliferation inhibition rate in GC insensitive group was lower than that in GC sensitive group (26.72%±21.82% vs 64.44%±25.48%, t=6.113, P<0.05). The PBMC proliferation inhibition rate in refractory group was lower than that in initial group (40.93%±28.57% vs 57.04%±31.19%, t=2.035, P=0.046). There was no statistical significance between gender, affected ear side, accompanied by vertigo or not, audiometric curve and the hearing improvement (χ² value was 2.320, 0.031, 2.143, 0.106, respectively, all P>0.05). Both in initial group and refractory group, the linear correlation analyses showed a significant positive correlation between PBMC proliferation inhibition rate and the PTA improvement (r value was 0.615, 0.657, respectively, all P<0.05), as well as a significant negative correlation between time period from onset to treatment and the PTA improvement(r value was -0.542, 0.370, respectively, all P<0.05). Conclusions The proliferation inhibition rate of PBMC in vitro by GC is correlated with patients′ hearing improvement. The proliferation inhibition test might be used to predict the sensitivity to GC treatment and be helpful for individualized treatment of SSNHLin clinical practice.