Title correlation analysis of chitinase-3 like-protein-1, matrix metalloproteinase-9 and imaging markers in middle aged and elderly patients with cerebral small vessel disease / 中华老年医学杂志

Objective:This study aimed to investigate the correlation between serum chitinase-3 like-protein-1(YKL-40), matrix metalloproteinase-9(MMP-9), and imaging markers in middle-aged and elderly patients with cerebral small vessel disease(CSVD).It also sought to elucidate the mechanism underlying the onset and development of CSVD.Methods:A case-control study was conducted with 130 CSVD patients and 20 age-matched controls admitted to the Department of Neurology of the Second Affiliated Hospital of Zhengzhou University between December 2020 and November 2022.Based on their white matter hyperintensity(WMH)and enlarged perivascular spaces(EPVS)scores, CSVD patients were categorized into mild, moderate, and severe groups.Cerebral microbleeds(CMBs)were classified into non-lacunar, lacunar, non-CMBs, and CMBs groups.Patients' general data were collected, and serum levels of YKL-40 and MMP-9 were measured using ELISA.Results:The results showed 34 cases in the severe WMH group, 35 cases in the EPVS severe group, 90 cases in the group with lacunar and 65 cases in the group with CMBs.Statistically significant differences(both P<0.05)in serum YKL-40 and MMP-9 in WMH severe groups compared to age control group.Statistically significant differences(both P<0.05)in serum YKL-40 and MMP-9 in EPVS severe groups compared to age control group.The increase of serum YKL-40 and MMP-9 levels was an independent risk factor for the severity of WMH( OR=1.25, 95% CI: 0.164-0.289, P<0.05)( OR=1.13, 95% CI: 0.090-0.160, P<0.05)and EPVS( OR=1.05, 95% CI: 0.021-0.076, P<0.05)( OR=1.02, 95% CI: 0.006-0.026, P<0.05).There was no significant different in the serum YKL-40 and MMP-9 levels between lacunar group and non-lacunar group( P>0.05).The serum YKL-40 and MMP-9 levels in the group with CMBs were significantly higher than those in the group without CMBs( P<0.05).The ROC curve showed that the optimal cutoff point of YKL-40 was 0.38, the area under the curve was 0.669(95% CI: 0.569-0.768, P<0.01), the optimal cutoff point of MMP-9 was 0.40, and the area under the curve was 0.746(95% CI: 0.644-0.848, P<0.01). Conclusions:Changes in serum YKL-40 and MMP-9 levels in middle-aged and elderly patients with CSVD were found to be associated with various imaging markers, potentially contributing to the onset and development of CSVD.These findings may aid in early CSVD diagnosis and provide insights for potential interventions.

Implementation of point-of-care platforms for rapid detection of porcine circovirus type 2

Background@#Porcine circovirus type 2 (PCV2) infection is ubiquitous around the world.Diagnosis of the porcine circovirus-associated disease requires clinic-pathological elements together with the quantification of viral loads. Furthermore, given pig farms in regions lacking access to sufficient laboratory equipment, developing diagnostic devices with high accuracy, accessibility, and affordability is a necessity. @*Objectives@#This study aims to investigate two newly developed diagnostic tools that may satisfy these criteria. @*Methods@#We collected 250 specimens, including 170 PCV2-positive and 80 PCV2-negative samples. The standard diagnosis and cycle threshold (Ct) values were determined by quantitative polymerase chain reaction (qPCR). Then, two point-of-care (POC) diagnostic platforms, convective polymerase chain reaction (cPCR, qualitative assay: positive or negative results are shown) and EZtargex (quantitative assay: Ct values are shown), were examined and analyzed. @*Results@#The sensitivity and specificity of cPCR were 88.23% and 100%, respectively; the sensitivity and specificity of EZtargex were 87.65% and 100%, respectively. These assays also showed excellent concordance compared with the qPCR assay (κ = 0.828 for cPCR and κ = 0.820 for EZtargex). The statistical analysis showed a great diagnostic power of the EZtargex assay to discriminate between samples with different levels of positivity. @*Conclusions@#The two point-of-care diagnostic platforms are accurate, rapid, convenient and require little training for PCV2 diagnosis. These POC platforms can discriminate viral loads to predict the clinical status of the animals. The current study provided evidence that these diagnostics were applicable with high sensitivity and specificity in the diagnosis of PCV2 infection in the field.

Study on injectable chitosan hydrogel with tendon-derived stem cells for enhancing rotator cuff tendon-to-bone healing / 中国修复重建外科杂志

OBJECTIVE@#To explore the effect of chitosan (CS) hydrogel loaded with tendon-derived stem cells (TDSCs; hereinafter referred to as TDSCs/CS hydrogel) on tendon-to-bone healing after rotator cuff repair in rabbits.@*METHODS@#TDSCs were isolated from the rotator cuff tissue of 3 adult New Zealand white rabbits by Henderson step-by-step enzymatic digestion method and identified by multidirectional differentiation and flow cytometry. The 3rd generation TDSCs were encapsulated in CS to construct TDSCs/CS hydrogel. The cell counting kit 8 (CCK-8) assay was used to detect the proliferation of TDSCs in the hydrogel after 1-5 days of culture in vitro, and cell compatibility of TDSCs/CS hydrogel was evaluated by using TDSCs alone as control. Another 36 adult New Zealand white rabbits were randomly divided into 3 groups ( n=12): rotator cuff repair group (control group), rotator cuff repair+CS hydrogel injection group (CS group), and rotator cuff repair+TDSCs/CS hydrogel injection group (TDSCs/CS group). After establishing the rotator cuff repair models, the corresponding hydrogel was injected into the tendon-to-bone interface in the CS group and TDSCs/CS group, and no other treatment was performed in the control group. The general condition of the animals was observed after operation. At 4 and 8 weeks, real-time quantitative PCR (qPCR) was used to detect the relative expressions of tendon forming related genes (tenomodulin, scleraxis), chondrogenesis related genes (aggrecan, sex determining region Y-related high mobility group-box gene 9), and osteogenesis related genes (alkaline phosphatase, Runt-related transcription factor 2) at the tendon-to-bone interface. At 8 weeks, HE and Masson staining were used to observe the histological changes, and the biomechanical test was used to evaluate the ultimate load and the failure site of the repaired rotator cuff to evaluate the tendon-to-bone healing and biomechanical properties.@*RESULTS@#CCK-8 assay showed that the CS hydrogel could promote the proliferation of TDSCs ( P<0.05). qPCR results showed that the expressions of tendon-to-bone interface related genes were significantly higher in the TDSCs/CS group than in the CS group and control group at 4 and 8 weeks after operation ( P<0.05). Moreover, the expressions of tendon-to-bone interface related genes at 8 weeks after operation were significantly higher than those at 4 weeks after operation in the TDSCs/CS group ( P<0.05). Histological staining showed the clear cartilage tissue and dense and orderly collagen formation at the tendon-to-bone interface in the TDSCs/CS group. The results of semi-quantitative analysis showed that compared with the control group, the number of cells, the proportion of collagen fiber orientation, and the histological score in the TDSCs/CS group increased, the vascularity decreased, showing significant differences ( P<0.05); compared with the CS group, the proportion of collagen fiber orientation and the histological score in the TDSCs/CS group significantly increased ( P<0.05), while there was no significant difference in the number of cells and vascularity ( P>0.05). All samples in biomechanical testing failed at the repair site during the testing process. The ultimate load of the TDSCs/CS group was significantly higher than that of the control group ( P<0.05), but there was no significant difference compared to the CS group ( P>0.05).@*CONCLUSION@#TDSCs/CS hydrogel can induce cartilage regeneration to promote rotator cuff tendon-to-bone healing.

Clinical Efficacy of Huanglian Jiedutang in Adjuvant Treatment of Acute Cerebral Infarction Complicated with Gastric Motility Disorder / 中国实验方剂学杂志

ObjectiveTo explore the clinical efficacy of Huanglian Jiedutang as an adjunctive treatment for acute cerebral infarction complicated with gastric motility disorder. MethodSixty patients with acute cerebral infarction complicated with gastric motility disorder with fire toxin syndrome were randomly divided into a western medicine control group （control group） and a traditional Chinese medicine （TCM） combined treatment group （observation group）， with 30 cases in each group. The control group received basic treatment for cerebral infarction and relevant western medical symptomatic treatment based on the patients' gastrointestinal symptoms. The observation group received Huanglian Jiedutang in addition to the treatment provided to the control group. The treatment course was 7 days. Neurological deficit scores and gastrointestinal dysfunction scores were assessed in both groups before treatment and on the 4th and 7th days of treatment. Gastrointestinal electrographic parameters， serum citrulline （CIT）， and motilin （MTL） levels were measured in both groups before treatment and on the 7th day of treatment. Clinical efficacy was compared between the two groups. ResultCompared with the baseline in both groups， the neurological deficit scores and gastrointestinal dysfunction scores were significantly reduced on the 4th and 7th days of treatment （P<0.05）. The reductions in these scores were more significant on the 7th day compared with those on the 4th day of treatment （P<0.05）. On the 4th and 7th days of treatment， the observation group showed a significantly greater reduction in neurological deficit scores and gastrointestinal dysfunction scores compared with the control group （P<0.05）. On the 7th day of treatment， compared with the baseline， both groups showed a significant increase in gastric antral and gastric body electric wave amplitudes as well as serum CIT and MTL levels （P<0.05）， and there were no statistically significant differences in the frequency of gastric antral and gastric body electric waves. On the 7th day of treatment， compared with the control group， the observation group had a significant increase in gastric antral and gastric body electric wave amplitudes as well as serum CIT and MTL levels （P<0.05）， and there were no statistically significant differences in the frequency of gastric antral and gastric body electric waves. After 7 days of treatment， the total effective rate in the observation group was 90.00% （27/30）， higher than 76.67% （23/30） in the control group， but the difference was not statistically significant. ConclusionAdjunctive treatment with Huanglian Jiedutang can effectively improve the symptoms of neurological function impairment and gastrointestinal dysfunction in patients with acute cerebral infarction complicated with gastric motility disorder， increase gastric antral and gastric body electric wave amplitudes， improve gastric motility disorder， and increase serum CIT and MTL levels， thereby improving the imbalanced secretion function of the gastrointestinal tract.

Fascia lata autograft bridging combined with long head of biceps tendon transposition for irreparable massive rotator cuff tear / 中华骨科杂志

Objective:To investigate the early clinical effect of fascia lata autograft bridging combined with the long head of biceps tendon transposition for treatment of irreparable massive rotator cuff tear.Methods:All of 31 cases of massive irreparable rotator cuff tear treated in our hospital from March 2016 to March 2020 were analyzed retrospectively. Among them, 17 cases (10 males, 7 females) were repaired with fascia lata autograft bridging under arthroscopy (patch group), the average age was 61.47±6.63 (ranging from 51 to 72) and 14 cases (4 males, 10 females) were repaired with fascia lata autograft bridging combined with the long head of biceps tendon transposition (combined group), the average age was 62.57±6.11 (ranging from 53 to 71). The operation time, intraoperative blood loss, postoperative complications, visual analogue scale (VAS) of pain before operation, at 1 week and 12 months after operation, Constant-Murley score of shoulder joint and American Association of shoulder and elbow Surgeons (ASES) score before operation, at 6 months and 12 months after operation were compared between the two groups. The outcome of rotator cuff healing was evaluated by MRI 1 year after operation.Results:All patients were followed up for 12-27 months (mean 18.33 ±6.8 months). There was no perioperative complication, and there was no significant difference in operation time between the two groups ( P>0.05) . The VAS score in the patch group was significantly higher than the combined group 1 week after operation ( t=2.09, P=0.048) , and there was no significant difference in VAS score 12 months after operation between the two groups. Constant-Murley score and ASES score in the combined group were significantly higher than the patch group at 6 months after operation ( t=5.23, P<0.001; t=4.45, P<0.001) , and there was no significant difference in Constant score and ASES score between the two groups at 12 months after operation. Constant score and ASES score in the two groups were significantly higher than those before operation. One year after operation, the MRI of the affected shoulder showed that the incidence of autograft patch thinning (Sugaya grade III) was 52.94%, the autograft patch structure failure rate (Sugaya grade IV and V) was 17.65% in the patch group, the autograft patch thinning rate (Sugaya grade III) was 35.71%, and the structural failure rate (Sugaya grade IV and V) was 7.14% in the combined group. The difference was statistically significant (χ 2=7.12, P=0.028) . Conclusion:Fascia lata autograft patch bridging combined with long head of biceps tendon transposition technique for treatment of irreparable massive rotator cuff tear has less pain 1 week after operation and better recovery of shoulder function half a year after operation. MRI showed better patch healing 1 year after operation.

Carbon Chain Length Determines Inhibitory Potency of Perfluoroalkyl Sulfonic Acids on Human Placental 3β-Hydroxysteroid Dehydrogenase 1: Screening, Structure-Activity Relationship, and In Silico Analysis / 生物医学与环境科学（英文）

OBJECTIVE@#This study aimed to compare 9 perfluoroalkyl sulfonic acids (PFSA) with carbon chain lengths (C4-C12) to inhibit human placental 3β-hydroxysteroid dehydrogenase 1 (3β-HSD1), aromatase, and rat 3β-HSD4 activities.@*METHODS@#Human and rat placental 3β-HSDs activities were determined by converting pregnenolone to progesterone and progesterone secretion in JEG-3 cells was determined using HPLC/MS-MS, and human aromatase activity was determined by radioimmunoassay.@*RESULTS@#PFSA inhibited human 3β-HSD1 structure-dependently in the order: perfluorooctanesulfonic acid (PFOS, half-maximum inhibitory concentration, IC 50: 9.03 ± 4.83 μmol/L) > perfluorodecanesulfonic acid (PFDS, 42.52 ± 8.99 μmol/L) > perfluoroheptanesulfonic acid (PFHpS, 112.6 ± 29.39 μmol/L) > perfluorobutanesulfonic acid (PFBS) = perfluoropentanesulfonic acid (PFPS) = perfluorohexanesulfonic acid (PFHxS) = perfluorododecanesulfonic acid (PFDoS) (ineffective at 100 μmol/L). 6:2FTS (1H, 1H, 2H, 2H-perfluorooctanesulfonic acid) and 8:2FTS (1H, 1H, 2H, 2H-perfluorodecanesulfonic acid) did not inhibit human 3β-HSD1. PFOS and PFHpS are mixed inhibitors, whereas PFDS is a competitive inhibitor. Moreover, 1-10 μmol/L PFOS and PFDS significantly reduced progesterone biosynthesis in JEG-3 cells. Docking analysis revealed that PFSA binds to the steroid-binding site of human 3β-HSD1 in a carbon chain length-dependent manner. All 100 μmol/L PFSA solutions did not affect rat 3β-HSD4 and human placental aromatase activity.@*CONCLUSION@#Carbon chain length determines inhibitory potency of PFSA on human placental 3β-HSD1 in a V-shaped transition at PFOS (C8), with inhibitory potency of PFOS > PFDS > PFHpS > PFBS = PFPS = PFHxS = PFDoS = 6:2FTS = 8:2FTS.

Gastrin attenuates sepsis-induced myocardial dysfunction by down-regulation of TLR4 expression in macrophages

Myocardial dysfunction is the most serious complication of sepsis. Sepsis-induced myocardial dysfunction (SMD) is often associated with gastrointestinal dysfunction, but its pathophysiological significance remains unclear. The present study found that patients with SMD had higher plasma gastrin concentrations than those without SMD. In mice, knockdown of the gastrin receptor, cholecystokinin B receptor (Cckbr), aggravated lipopolysaccharide (LPS)-induced cardiac dysfunction and increased inflammation in the heart, whereas the intravenous administration of gastrin ameliorated SMD and cardiac injury. Macrophage infiltration plays a significant role in SMD because depletion of macrophages by the intravenous injection of clodronate liposomes, 48 h prior to LPS administration, alleviated LPS-induced cardiac injury in Cckbr-deficient mice. The intravenous injection of bone marrow macrophages (BMMs) overexpressing Cckbr reduced LPS-induced myocardial dysfunction. Furthermore, gastrin treatment inhibited toll-like receptor 4 (TLR4) expression through the peroxisome proliferator-activated receptor α (PPAR-α) signaling pathway in BMMs. Thus, our findings provide insights into the mechanism of the protective role of gastrin/CCKBR in SMD, which could be used to develop new treatment modalities for SMD.

Analysis on prevalence and epidemic risk of animal plague in different ecological plague foci in Inner Mongolia Autonomous Region / 中华预防医学杂志

The risk of plague epidemics and relapse of various types of plague foci persists in Inner Mongolia Autonomous Region. For Marmota sibirica plague foci, the animal plague has not been found but antibody has been detected positive. Nowadays, Marmota sibirica has been increasing in population and distribution in China. In bordering countries Mongolia and Russia, the animal plague has been continuously prevalent. For Spermophilus dauricus plague foci, the animal plague has been taken place now and then. Compared to the above foci, the animal plague is most prevalent in Meriones unguiculatus plague foci and frequently spread to humans. Due to higher strain virulence and historical disaster in Marmota sibirica plague foci and Spermophilus dauricus plague foci, plague prevention and control should be strengthened on these foci. In addition to routine surveillance, epidemic dynamics need to be further monitored in these two foci, in order to prevent their relapse and spread to humans.

Youguiwan Regulates CXCL8/CXCR1/2 Signaling Pathway and Ang-1 and Ang-2 Expression to Promote Ovarian Angiogenesis in Rats / 中国实验方剂学杂志

ObjectiveTo study the effects of Chinese herbal compound Youguiwan on angiogenesis of rats with ovarian dysfunction caused by natural aging and its relationship with chemokine interleukin 8 （CXCL8）/CXC chemokine receptor 1/2 （CXCR1/2） signaling pathway， angiopoietin 1 （Ang-1）， and angiopoietin 2 （Ang-2）， so as to explore its mechanism in improving the ovarian function. MethodFifty six female SD rats were randomly divided into the young control group （n=8） and modeling group （n=48， ovarian dysfunction caused by natural aging）. Rats in both the young control and modeling groups were routinely fed， during which the ones in the modeling group underwent exfoliative cytology of vaginal smears for five to seven days. The ones presented with prolonged estrous cycle， followed by continuous estrus and repeated pseudopregnancy revealed by vaginal cytology during four consecutive estrous cycles indicated early aging， and the young rats with keratinocyte proliferation index higher than 50% for 10 consecutive days were classified into the young control group. The successfully modeled rats were randomly divided into the early-aged group， estrogen （65 μg·kg-1·d-1） group， Zuoguiwan （33 g·kg-1·d-1） group， as well as the low-， medium-， and high-dose （1.2， 2.4， 4.8 g·kg-1·d-1） Youguiwan groups. Rats in the young control group and the early-aged group were gavaged with the same volume of normal saline for 30 days. After the experiment， the morphological changes in rat ovary were observed by hematoxylin-eosin （HE） staining. The protein expression levels of chemokines CXCL8， CXCR1， CXCR2， Ang-1， and Ang-2 in rat ovary were detected by Western blotting and immunohistochemistry， and the mRNA expression levels of CXCL8， CXCR1， CXCR2， Ang-1， and Ang-2 by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the young control group， the early-aged group exhibited reduced number of growing follicles， corpus luteum， and blood vessels at all levels， elevated atretic follicles （P<0.01）， up-regulated protein and mRNA expression of CXCL8， CXCR1， and CXCR2 in the ovarian tissue （P<0.01）， and down-regulated Ang-1 and Ang-2 protein and mRNA expression （P<0.05）. Compared with the early-aged group， each medication remarkably increased the number of growing follicles， corpus luteum， and blood vessels （P<0.05）， lowered the number of atretic follicles （P<0.05）， down-regulated the protein and mRNA expression levels of CXCL8， CXCR1， and CXCR2 in the ovarian tissue （P<0.05）， and up-regulated the protein and mRNA expression levels of Ang-1 and Ang-2 （P<0.05）. ConclusionYouguiwan down-regulates the levels of CXCL8， CXCR1， and CXCR2 in rat ovary and up-regulates the levels of Ang-1 and Ang-2 to promote ovarian angiogenesis and improve rat ovarian function.

Study on Liver Injury of Different Components of Evodia Fructus in Vivo and in Vitro / 中国实验方剂学杂志

ObjectiveTo evaluate the in vivo and in vitro toxicity of Evodia Fructus water extraction and its index components， and provide a basis for basic research on the toxic substances of Evodia Fructus. MethodInstitute of Cancer Research（ICR） mice were divided into high， medium and low dose groups of water extraction of Evodia Fructus and a blank control group. The administration groups were respectively given 80，60，40 g·kg-1 water extraction of Evodia Fructus， the blank control group was given distilled water in equal volume， blood was taken 24 hours later to determine the serum alanine aminotransferase（ALT）and aspartate aminotransferase（AST）values， the liver was weighed and histopathological examination was performed. Evodia Fructus water extract， evodiamine， rutaecarpine and limonin were respectively acted on HepG2 cells for 24 h， and cell counting kit-8（CCK-8） method was used to investigate the cytotoxicity. The ICR Mice were divided into two groups， one group was given by oral gavage and the other group was given intraperitoneal injection. The two routes of administration were separately given 3 index components of Evodia Fructus， and the dosage was 200 mg·kg-1. Take blood 24 hours after administration to determine the activity of ALT and AST in serum， and take liver to calculate liver index. ResultCompared with the blank group， the high and medium dose groups of Evodia Fructus water extract were depressed 24 hours after administration， and the behavior of the low dose group was not significantly abnormal. The serum biochemical results showed that the activities of serum ALT and AST in the high and medium dose groups were significantly increased （P<0.01）， the activities of serum ALT and AST in the low dose group were significantly increased， and the histopathological results showed that the high and medium dose groups were significantly increased Punctate necrosis and vacuolar degeneration appeared in the liver of the medium dose group， and there was no obvious abnormality in the low dose group. Compared with the blank group， evodiamine and rutaecarpine had a certain inhibitory effect on the proliferation of HepG2 cells， but the inhibitory effect was not strong. Limonin had no significant inhibitory effect on the proliferation of HepG2 cells. Compared with the control group， the 3 index components of Evodia Fructus have no effect after oral administration. There was no significant difference in the activity of ALT and AST in serum of mice， and there was no significant difference in liver index. Intraperitoneal injection of evodiamine and rutaecarpine can cause the activity of serum ALT and AST to increase， and limonin can cause ALT activity was significantly increased （P<0.01）， and the liver index was significantly increased （P<0.05）. ConclusionEvodia Fructus water extract can cause acute liver injury in mice， Oral administration of evodiamine， rutaecarpine and limonin had no damage to the liver of mice. Intraperitoneal administration of evodiamine and rutaecarpine had no effect on liver injury in mice， and intraperitoneal administration of limonin could cause acute liver injury in mice.

The preliminary exploration of Calu-3 cell air-liquid interface culture and exposure research / 中国药理学通报

Aim To explore the air-liquid interface(ALI)culture conditions of Calu-3 cells and their tolerance to exposure to clean air in the exposure system. Methods Calu-3 cells were cultured in three stages:flask expansion,liquid-liquid culture until full Transwell membrane covered,and ALI culture,and the cell barrier status was determined by cell trans-epithelium electrical resistant(TEER)measurement and expression of tight junction protein ZO-1; Calu-3 cells were exposed to clean air using the air-liquid interface exposure system for 1,2,3,4,5,6 hours,and cells cultured in incubators were used as controls to detect changes in Calu-3 cell activity and TEER after exposure,and to determine the single maximum exposure time of Calu-3 cells in the in vitro exposure system based on ALI culture. Results Calu-3 cells were cultured in liquid-liquid culture for 8±1 days to grow full Transwell membranes and barrier formation,and then were transferred to ALI culture. The barrier was intact and in a stable state for 1 to 18 days of ALI culture,and could be used for exposure experiments. The activity of Calu-3 cells decreased significantly after 6 hours of clean air exposure(P<0.01),and TEER decreased significantly after 4,5,and 6 hours of clean air exposure(P<0.05,P<0.01,P<0.01). Conclusions Calu-3 cells cultured for 1 to 18 days in ALI can be used for air-liquid interface exposure experiments; the combined changes in cell activity and TEER suggest that the maximum single exposure time for Calu-3 cells exposed to clean air in the exposure system is 3 hours.

Interlaboratory method validation of slope ratio determination for anticoagulant activity of leeches / 中国药理学通报

Aim To investigate the slope ratio method for the determination of anticoagulant activity of leeches. Methods Three batches of leeches, four groups of Japanese medical vermiculite yinpian and fifteen groups of leech preparations were chosen, with contrast medicinal leeches herbs and Philippine cattle leech contrast medicinal materials, and different concentrations of leaching solutions were prepared in parallel. APTT value was determined after anticoagulant activity was determined by slope ratio method for the joint validation of laboratory, intermediate precision and accuracy between the linear range. Results The slope ratio method was accurate and accurate in the determination of anticoagulant activity of leeches, with linearity between 64% and 156% relative titer level. Conclusion Slope ratio method can be used to determine the anticoagulant activity of leeches.

Study on pharmacodynamics and pharmacokinetics of Phytolacca acinosa Roxb.And its mechanism / 中国药理学通报

Aim To explore the diuretic effect, diuretic mechanism and pharmacokinetics of Phytolacca acinosa Roxb., and clarify its "quantity-time-effect" relationship.Methods Firstly, qualified rats were modeled by water load model, given different doses of Phytolacca acinosa Roxb.aqueous extract, then the diuretic effect was investigated.Secondly, Western blot was used to detect the protein expression of aquaporins AQP2, AQP4 and the angiotensin II receptors ATGR1, ATGR2, and renin in the RAAS system in kidney tissues.Thirdly, the established LC-MS/MS biological analysis method was used to detect the esculentoside A(EsA)content in the plasma, calculate the pharmacokinetic parameters and analyze the correlation between the blood concentration and the drug effect.Results The water load model was successfully established.Compared with the model group, hydrochlorothiazide had a significant diuretic effect(P<0.01).Low, medium and high dose groups of Phytolacca acinosa Roxb.all had obvious diuretic effects(P<0.01), EsA also had a significant diuretic effect(P<0.05).Phytolacca acinosa Roxb.aqueous extract and EsA significantly down-regulated the expression of AQP2, AQP4, ATGR1 and renin protein.The pharmacokinetic results showed that the Cmax and AUC0-t of EsA in the plasma of rats in the low, medium, and high dose groups of aqueous extract increased with the increase of the dose.Conclusions Phytolacca acinosa Roxb.had a diuretic effect, which is related to inhibiting the expression of aquaporins AQP2 and AQP4 and inhibiting the expression of angiotensin II type 1 receptor and renin, thereby inhibiting the reabsorption of renal tubules and collecting ducts.

Regulation Effect and Clinical Efficacy of Addition and Subtraction Therapy of Jinkui Shenqiwan Combined with Buzhong Yiqitang on Immune Inflammatory Factors of Patients with Postmenopausal Osteoporosis and Deficiency of Spleen and Kidney / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of addition and subtraction therapy of Jinkui Shenqiwan combined with Buzhong Yiqitang to postmenopausal osteoporosis （PMO） with deficiency of spleen and kidney， and to investigate its regulation effect on immune inflammatory factors. Method:One hundred and sixty patients were randomly divided into observation group and control group， with 80 cases in each group. Both groups got comprehensive western medicine treatment measures. Patients in control group additionally got Zhuanggu Zhitong capsule， 4 capsules/time， 3 times/day. Patients in observation group additionally got addition and subtraction therapy of Jinkui Shenqiwan combined with Buzhong Yiqitang， 1 dose/day. The treatment was continued for 24 weeks. Before and after treatment， lumbar L2-4 bone mineral density （BMD） was detected by Dual energy X-ray absorptiometry （DXA） and lumbar BMD was detected by quantitative CT （QCT）. Scores of traditional Chinese medicine（TCM） syndromes and Chinese osteoporosis-targeted quality of life questionnaire （COQOL） were graded. Levels of Estradiol （E₂）， type Ⅰ procollagen amino terminal pro peptide （PINP）， serum osteocalcin （OC）， osteoprotegerin （OPG）， type Ⅰ collagen cross-linked C-terminal peptide （S-CTX）， tartrate resistant acid phosphatase （TRACP） and urinary pyridinoline （PYD） were detected. Levels of CD4⁺ T cells， CD8⁺ T cells， interleukin-17 （IL-17）， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， <italic>γ-</italic>interferon（IFN-<italic>γ</italic>） and interleukin-4 （IL-4） were calculated. The proportion of T helper cell （Th）17 and regulatory T cell （Treg） in CD4⁺ T cells was calculated. Besides， the safety was evaluated. Result:Bone density was detected by DXA in observation group， and its T-value and bone density detected by QCT were all higher than those in control group （<italic>P</italic><0.01）. After treatment， scores of TCM syndrome and COQOL were lower than those in control group （<italic>P</italic><0.01）. Levels of PINP， OC， S-CTX， TRACP and PYD/Cr were all lower than those in control group （<italic>P</italic><0.01）. Levels of OPG， CD8⁺ and Treg were higher than those in control group （<italic>P</italic><0.05）， levels of Th17， Th17/Treg， CD4⁺/CD8⁺， IL-17， TNF-<italic>α</italic> and IFN-<italic>γ </italic>were lower （<italic>P</italic><0.01）， and levels of IL-4 and E₂ were higher than those in control group （<italic>P</italic><0.01）. The clinical efficacy in observation group was better than that in control group （<italic>Z</italic>=2.103， <italic>P</italic><0.05）. Conclusion:On the basis of calcium and vitamin D supplementation， Jinkui Shenqiwan combined with Buzhong Yiqitang can improve levels of E₂ and bone density， reduce clinical symptoms， improve quality of life， regulate bone metabolism index and immune inflammation reaction， with better clinical efficacy and safety.

Culture and differentiation of mouse tracheal epithelial cells at air-liquid interface / 中国药理学通报

Aim Air-liquid interface (ALI) cultures of mouse tracheal epithelial cells (MTEC) are a well-established model to study airway epithelial cells. MTEC provides a powerful ap¬proach for the evaluation of the inhalation toxicological in vitro. Methods C57BL/6 mouse tracheal-bronchial epithelial cells were obtained by digestion with protease in cold temperature o- vemight, and the digestion time was optimized to ensure the quantity and viability of the obtained cells. The cells were cul¬tured into collagen coated Transwell inserts. Proliferating phase and air-liquid interface culture were promoted with different cul¬ture media. The expression of tight junction protein and cell trans-epithelial electrical resistance(TEER) were used to evalu¬ate the formation of tight junction between cells and the analysis of cell polarity. The cilia structure was confirmed by electron mi¬ croscopy and immunofluorescence. Results Highly purified and viable primary airway epithelial cells could be harvested and subcultured by our methods, including morphology and immuno- cytochemistry staining confirmed the expression of MUC5AC, a- tubulin, p-tubulin-IV and ZO-1. The development of tight-junc¬tions and epithelium were similar with pseudostratified ciliated columnar epithelium morphology. Conclusions A comprehen¬sive protocol for ALI culture was established, reproducing the characteristic pseudostratified ciliated columnar epithelium mor¬phology and physiological functions in vitro. The MTEC protocol provides a stable and reliable method for the isolation, mucocili¬ary differentiation and reproducing.

Effect of infiltrating nasal packing with local anesthetics in postoperative pain and anxiety following sinonasal surgeries: a systemic review and meta-analysis / Efeito do tamponamento nasal infiltrado com anestésicos locais na dor e ansiedade pós-operatórias após cirurgias nasosinusais: revisão sistêmica e metanálise

Abstract Introduction: Packing of the nasal cavity has traditionally been used for postoperative bleeding control and decreasing synechia formation in patients undergoing nasal surgeries. Although absorbable nasal packing has been gaining popularity in the recent years, nonabsorbable nasal packing is still often used in nasal surgeries in various parts of the world. It is known to be associated with pain and discomfort especially upon and during removal, and previous reviews have only evaluated the effects of local anesthetic infiltration of nasal packing in septal surgeries. Objective: To evaluate the effect of infiltrating nasal packing with local anesthetics in postoperative pain and anxiety following sinonasal surgeries Materials and methods: We searched the PubMed and Embase databases from their earliest record to April 27, 2019, randomized controlled trials and prospective controlled trials for review, and included only randomized controlled trials for data analysis. We included studies using topical anesthetics-infiltrated nasal packing following sinonasal surgeries and evaluated the effectiveness compared to placebo packing in pain reduction during postoperative follow up, as well as the effectiveness in anxiety reduction. Results: Among 15 studies included for review, 9 studies involving 765 participants contributed to the meta-analysis. In terms of pain reduction, our analysis showed significant standard mean differences regarding effectiveness at postoperative 1, 12, 24 h interval for all surgical groups combined, in the sinus surgery group, as well as during nasal packing removal. There was no consistent evidence to support the effectiveness in anxiety reduction. Conclusions: Our study supports anesthetics infiltration of nasal packing as an effective method in managing pain in patients with nasal packing after sinonasal surgeries. However, the level of evidence is low. More high-quality randomized controlled trials are needed to establish its effectiveness in reducing anxiety. We believe this review is of great clinical significance due to the vast patient population undergoing sinonasal surgeries. Postoperative local hemorrhage remains the greatest concern for ear nose and throat surgeons due to the rich vasculature of the nose and sinuses. Sinonasal packing provides structural support and serves as an important measure for hemostasis and synechia formation. Although absorbable packing has been gaining popularity in the recent years, nonabsorable packing materials are still used in many countries due to lower cost. Infiltration of nasal packing with local anesthetic provides a solution to the discomfort, nasal pressure and nasal pain experienced commonly by the patients as evidenced by our analysis.

Resumo Introdução: O tamponamento da cavidade nasal tem sido usado tradicionalmente para controle do sangramento pós-operatório e diminuição da formação de sinéquia em pacientes submetidos a cirurgias nasais. Embora o tamponamento nasal absorvível tenha ganhado popularidade nos últimos anos, o tampão nasal não absorvível ainda é frequentemente usado em várias partes do mundo. Sabe-se que o tamponamento está associado a dor e desconforto, especialmente na sua remoção, e revisões anteriores avaliaram apenas os efeitos do tampão com anestésico local em cirurgias do septo nasal. Objetivo: Avaliar o efeito do tamponamento nasal infiltrado com anestésicos locais na dor e ansiedade pós-operatórias após cirurgias nasosinusais. Material e métodos: Para a revisão, pesquisamos nos bancos de dados PubMed e Embase desde o registro mais antigo até 27 de abril de 2019, incluímos ensaios clínicos controlados e randomizados, a ensaios clínicos prospectivos controlados e apenas ensaios clínicos controlados e randomizados para análise de dados. Incluímos estudos que usaram tamponamento nasal infiltrado com anestésicos tópicos após cirurgias nasosinusais e avaliamos a eficácia em comparação com o tamponamento com placebo na redução da dor durante o acompanhamento pós-operatório, bem como os efeitos na redução da ansiedade. Resultados: Entre os 15 estudos incluídos, 9, que envolveram 765 participantes, contribuíram para a metanálise. Em termos de redução da dor, nossa análise mostrou diferenças médias padrão significantes em relação à eficácia no pós-operatório nos intervalos de 1, 12, 24 horas para todos os grupos cirúrgicos combinados, no grupo da cirurgia sinusal e durante a remoção do tamponamento nasal. Não houve evidências consistentes para apoiar a eficácia na redução da ansiedade. Conclusões: Nosso estudo apoia o uso de tamponamentos nasais infiltrados com anestésicos locais como um método eficaz no tratamento da dor em pacientes após cirurgias nasosinusais. No entanto, o nível de evidência é baixo. São necessários mais ensaios clínicos randomizados de alta qualidade para estabelecer sua eficácia na redução da ansiedade. Acreditamos que esta revisão seja de grande significado clínico devido à vasta população submetida a cirurgias nasosinusais. A hemorragia local pós-operatória continua a ser a maior preocupação para os cirurgiões otorrinolaringológicos devido à rica vasculatura do nariz e seios nasais. O tamponamento nasosinusal fornece suporte estrutural e serve como uma medida importante para a hemostasia e formação de sinéquias. Embora o tamponamento absorvível tenha ganhado popularidade nos últimos anos, os materiais de tamponamento não absorvíveis ainda são usados em muitos países devido ao menor custo. A infiltração do tamponamento nasal com anestésicos locais fornece uma solução para desconforto, pressão e dor nasal comumente referida pelos pacientes, como evidenciado por nossa análise.

Performance of Posture Control for Professional Dancers / 中国康复理论与实践

Objective:To explore the posture control of professional dancers. Methods:From April to August, 2017, 21 professional dancers from an international famous club were as experimental group. Matching the height and age, etc., 21 medical workers were recruited as control group. They were tested with Tetrax Balance Evaluation System, and assessed with Trunk Stability Test (TST) and the Star Excursion Balance Test (SEBT). Results:There was no significant difference on fail-time of TST and maximum distance of SEBT between right and left sides in both groups. The fail-time of TST was less in the experimental group than in the control group (t =-2.667, P < 0.05), as well as the maximum distance of SEBT (t = -3.991, P < 0.001). There was no significant difference on falling index between both groups (t = 1.810, P > 0.05). Conclusion:Compared with medical workers, professional dancers do better in static balance, but worse in dynamic balance. Their performance of posture control is almost the same as the others.

Effect of Jingui Shenqiwan Combined with Fracture Three-stage Syndrome Differentiation on Recovery of Patients with Osteoporotic Vertebral Compression Fractures After Operation / 中国实验方剂学杂志

Objective:To evaluate the short-term and long-term efficacy of Jingui Shenqiwan combined with three-stage syndrome differentiation on osteoporotic vertebral compression fractures (OVCFs) after operation, and its regulatory effect on biomarkers of bone metabolism. Method:One hundred and thirty-five patients were randomly divided into control group (67 cases) and observation group (68 cases) by random number table. A total of 58 patients in control group completed the treatment (4 patients were exfoliated, 2 patients were lost to follow-up, 3 patients were eliminated); and 60 patients in observation group completed the treatment (3 patients were exfoliated, 2 patients were lost to follow-up, 3 patients were eliminated). Both groups patients were given calcitonin injection through intramuscular injection, 20 u/time, 1 time/week, for 12 weeks, calcium carbonate D3 chewable tablets (Ⅱ), 1 tablet/time, 2 times/day, and alendronate sodium tablets, 70 mg/time, 1 time/week. Patients in control group got Bushen Jiangu capsule, 4 grains/time, 3 times/day. And patients in observation group got modified Jingui Shenqiwan combined with fracture three-stage symptom differentiation, 1 dose/day. The courses of treatment in the two groups were 24 weeks, and a 24 week follow-up was provided. Before the operation and at the 12th and 24th week after operation, the short-term efficacy indexes, such as back pain, lumbar function, traditional Chinese medicine （TCM） syndromes and complications, were recorded. And the long-term efficacy indexes, such as recovery of responsible vertebral body, lumbar function, bone density and quality of life and incidence of 48 week re-fracture, were also recorded. Before and after operation, levels of bone alkaline phosphatase (BALP), osteocalcin (BGP), tartrate resistant acid phosphatase-5b (TRAP-5b), type I collagen carboxy terminal prepeptide (PICP), type I collagen cross-linked C-terminal peptide (β-CTX) and N-MID-OT were detected, and the safety was evaluated. Result:The comprehensive efficacy in observation group was superior to that in control group (Z=2.026, P<0.05). At the 12th and 24th week after operation, scores of back pain, lumbar function and TCM syndromes were all lower than those in control group (P<0.01), and score of lumbar function at the 48th week after operation was also lower than that in control group (P<0.01). Bone density was higher than that in control group at the 24th and 48th week after operation, and score of quality of life was lower than that in control group (P<0.01). At the 24th and 48th week after operation, Cobb angles were less than those in control group, and heights of responsible centrums (anterior, central, posterior) were higher than those in control group. Cumulative incidence of complications in control group was 51.72% (30/58), which was higher than 26.67% (16/60) in control group (χ2=7.784, P<0.01). The levels of BGP were higher than those in observation group at the 24th and 48th week after operation, and the levels of BALP, TRAP-5b, PICP, β-CTX and N-MID-OT were all lower than those in control group (P<0.01). And there was no side effect relating to Jingui Shenqiwan. Conclusion:Modified Jingui Shenqiwan combined with fracture three-stage symptom differentiation can reduce the symptoms of back pain and promote the recovery of lumbar function, with a significant short-term comprehensive efficacy. In the long term, it can improve the strength of responsible centrums, restore the anatomical structure of injured centrums, increase the bone density of centrums, further improve the lumbar function, reduce the occurrence of complications, regulate the markers of bone metabolism, and improve osteoporosis.

Trichostatin A inhibits the proliferation of gastric cancer cells through the promotion of autophagy via p-Akt/p-mTOR pathway / 医学研究生学报

Objective　Trihostatin A (TSA) is a histone deacetylase inhibitor of oxime salts, which has certain anti-tumor activity. This article mainly investigates the molecular mechanism of TSA inhibiting cell proliferation through p-AKT/p-mTOR pathway in gastric cancer cells.　Methods　Gastric cancer cell line-SGC-7901 were treated with TSA of different concentrations, and the inhibition rate of TSA on the cells was detected by MTT assay. The cells were divided into control group (without any treatment), TSA-treated group (200ng/ml TSA), p-AKT covering group (200 ng/mL TSA+8 μg/mL SC79) and autophagy inhibition group (5 mmol/mL 3-methyladenine+200 ng/mL TSA). The protein expression distribution of Lc3 in control and TSA group were detected by cell immunofluorescence staining. The relative expression levels of p-AKT, p-mTOR and autophagy related proteins Lc3 and P62 in control group, TSA group and p-AKT covering group were detected by Western blot. The proliferation of cells in control group, TSA group, p-AKT covering group and autophagy inhibition group were measured by EdU and cell count assay.　Results　After 24h of treatment, Lc3 in TSA group formed a large number of granular aggregates in the cytoplasm, and the fluorescence distribution changed from the initial diffuse to dense. The TSA group showed a significant reduction in green fluorescence compared with the control group in the EdU experiment. The expression levels of p-AKT in the control group, TSA group and the autophagy inhibition group were 1.78±0.19, 0.92±0.03 and 1.71±0.19, respectively, and Lc3 were 0.21±0.01, 0.79±0.06 and 0.55±0.10, respectively. Compared with the control group, the relative expression level of p-AKT in the TSA group all decreased, while the expression level of Lc3 increased (P <0.05). p-mTOR in the three groups was 0.80±0.16, 0.45±0.04 and 0.98±0.16, respectively. Compared with the control group, the relative expression level of p-mTOR in the TSA group all decreased (P <0.05). P62 in the three groups was 1.17±0.15, 0.48±0.08 and 0.77±0.10, respectively. Compared with the control group, the relative expression level of P62 in the TSA group all decreased (P<0.05). Compared with the TSA group, p-AKT, p-mTOR and P62 expression in the p-AKT covering group increased (P<0.05), while Lc3 expression decreased (P<0.05). Compared with the control group, the inhibition effect of cell growth curve was the most obvious in the TSA group, while the cell growth curve of p-AKT covering group and autophagy inhibition group showed a partial recovery compared with the TSA group.　Conclusion　TSA can promote autophagy by inhibiting p-AKt/p-mTOR pathway, thus inhibiting the proliferation of gastric cancer cells.

RON and MET Co-overexpression Are Significant Pathological Characteristics of Poor Survival and Therapeutic Targets of Tyrosine Kinase Inhibitors in Triple-Negative Breast Cancer / Journal of the Korean Cancer Association, 대한암학회지

Purpose@#Triple-negative breast cancer (TNBC) is highly malignant and has poor prognosis and a high mortality rate. The lack of effective therapy has spurred our investigation of new targets for treating this malignant cancer. Here, we identified RON (macrophage-stimulating 1 receptor) and MET (MET proto-oncogene, receptor tyrosine kinase) as a prognostic biomarker and therapeutic targets for potential TNBC treatment. @*Materials and Methods@#We analyzed RON and MET expression in 187 primary TNBC clinical samples with immunohistochemistry. We validated the targeted therapeutic effects of RON and MET in TNBC using three tyrosine kinase inhibitors (TKIs): BMS-777607, INCB28060, and tivantinib. The preclinical therapeutic efficacy of the TKIs was mainly estimated using a TNBC xenograft model. @*Results@#Patients with TNBC had widespread, abnormal expression of RON and MET. There was RON overexpression, MET overexpression, and RON and MET co-overexpression in 63 (33.7%), 63 (33.7%), and 43 cases (23.0%), respectively, which had poor prognosis and short survival. In vivo, the TKI targeting RON ant MET inhibited the activation of the downstream signaling molecules, inhibited TNBC cell migration and proliferation, and increased TNBC cell apoptosis; in the xenograft model, they significantly inhibited tumor growth and shrank tumor volumes. The TKI targeting RON and Met, such as BMS-777607 and tivantinib, yielded stronger anti-tumor effects than INCB28060. @*Conclusion@#RON and MET co-overexpression can be significant pathological characteristics in TNBC for poor prognosis. TKIs targeting RON and MET have stronger drug development potential for treating TNBC.