Objective To establish a RP-HPLC method for determining the content of puerarin in Xinkeshu Tablets. Method The analysis was carried on a column of Zorbax Eclipse XDB-C_(18)(4.6 mm?150mm,5?m)at 25℃,with methanol-water-phosphoric acid(23:77:0.5)as mobile phase.The flow rate was 1.0 mL?min~(-1),and the detection wavelength was 250 nm.Result A good linearity was found in the range of 27.1～271.0 ng.The average recovery rate was 100.5 %,and RSD was 1.8%(n=5).Conclusion The method is simple,rapid,accurate,reproducible,and can be used to control the quality of Xinkeshu Tablets.

Objective To observe the effects of melatonin on synaptic plasticity impaired by spinal cord injury in rats. Methods A total of 54 female Sprague-Dawley rats were divided into sham group (n=18), control group (n=18) and melatonin group (n=18). Spinal cord inju-ry model was established with modified Allen's method at T10 (10 g from 25 mm height). The number of neurons and the expression of the Nissl body were detected with immunofluorescence and Nissl staining. The expression of neurofilament-200 (NF-200), brain-derived neuro-trophic factors (BDNF), Synapsin I and growth-associated protein-43 (GAP-43) was detected with Western blotting. Results Seven days af-ter injury, the number of motoneurons, the expression of Nissl body in motoneurons, and the expression of BDNF, Synapsin I and GAP-43 decreased in the control group compared with those in the sham group, and they increased in the melatonin group compared with those in the control group. Conclusion Melatonin can repair the impaired synaptic plasticity, which might promote the functional recovery after spi-nal cord injury.

OBJECTIVE To analyze the antimicrobial activity of recombinant human ?-defensin 3(rhBD-3) on clinically isolated multidrug-resistant bacterial strains.METHODS The antimicrobial activity of rhBD-3 on clinically isolated multidrug-resistant Staphylococcus aureus,Enterococcus faecium,Acinetobacter baumannii and Pseudomonas aeruginosa from the wards of burns department was measured by turbidity method.RESULTS rhBD-3 Demonstrated antimicrobial activity against all the strains in a dose-dependent manner.The minimal inhibitory concentration(MIC) to Gram-positive strains and Gram-negative strains was 4 ?g/ml and 8 ?g/ml,respectively.CONCLUSIONS rhBD-3 Has significant antimicrobial activity against clinically isolated multidrug-resistant strains and thus implies therapeutic potential as an effective substitute for the present drug-resistant bacteria.

Objective To analyze the association between the polymorphism of 5509-5511delCAA in exon4 of Tim-1 gene and the susceptibility to rheumatoid arthritis(RA)in a group of Han population from Yunnan province.Methods The PCR-DNA se-quencing analysis was used to detect the polymorphism of 5509-5511delCAA in exon4 of Tim-1 gene in 196 RA Han nationality pa-tients and 190 individuals of healthy physical examination(control).The ELISA method and indirect immunofluorescence assay was adopted to detect the three related aotuantibodies of RA,including the rheumatoid factors(RF)and the anti-cyclic citrullinated pep-tide(Anti-CCP)and the anti-keratin antibodies(AKA).Results There were statistically significant differences in genotype and al-lele frequency of 5509-5511delCAA between the RA group and the control group(P <0.05);the positive rate of RF and the anti-CCP between different genotypes of 5509-5511delCAA in the RA group had no statistically significant differences(P >0.05 ),but the positive rate of AKA between different genotypes of 5509-5511delCAA in the RA group had statistically significant differences (P <0.05).Conclusion The 5509-5511delCAA site of Tim-1 exon4 has the polymorphic variation,the 5509-5511delCAA genotype maybe related with the genetic susceptibility to RA in Han population of Yunnan province,the different genotypes does not affect the expression of RF and the anti-CCP,but affects the expression of AKA.

Objective To investigate and analyze the applications of liver grafts from donation after death children (≤5 kg) in pediatric liver transplantation.Methods A retrospective analysis was conducted on 18 cases of pediatric recipients using liver grafts from donation after death children (≤5 kg) during January 2013 to December 2015.The survival rate of patients,recovery of liver function,incidence of lung infection,vascular complications and biliary complications were observed,and the causes of deaths were analyzed.Results The median follow-up time of all the patients was 28.4 months (0.06-52.2 months).The 1-and 3-year cumulative survival rate was the same 88.9％,the incidence of lung infection was 22.2％,of vessel complications was 33.3％,and biliary complication rate was 11.1％.Two cases died after liver transplantation.Conclusion The applications of liver donation after death children (≤5 kg) were satisfactory,but there is still relatively limited experience,and further observation and research are needed.

Objective:To investigate the expressions of apoptosis-related factors survivin, p53 and human epidermal growth factor receptor 2 (HER2) in breast cancer tissues and their prognostic value.Methods:A total of 131 patients undergoing radical mastectomy for breast cancer who were admitted to Tangshan Maternal and Child Health Care Hospital from February 2015 to January 2019 were selected as the research subjects. During the operation, the cancer tissues and adjacent tissues (normal tissues >3 cm from the tumor margin) were collected from the patients. Expressions of survivin, p53 and HER2 in cancer tissues and adjacent tissues of patients were detected by using immunohistochemistry. The prognoses of patients were recorded after the follow-up for 3 years; the recurrence, metastasis and death treated as the poor prognosis, the rest prognoses of patients were treated as the good prognosis group. The difference of clinicopathological characteristics between the poor prognosis group and the good prognosis group was compared. Multivariate logistic regression was used to analyze risk factors for prognosis of breast cancer patients. The result of prognosis of breast cancer was taken as the golden standard. The receiver operating characteristic (ROC) curve was used to analyze the value of survivin pasitive, p53 pasitive, HER2 pasitive alone, the combination of both and the combination of the there in the judgement of poor prognosis of breast cancer.Results:The positive expression rates of survivin [49.6% (65/131) vs. 7.6% (10/131)], p53 [60.3% (79/131) vs. 13.0% (17/131)] and HER2 [79.4% (104/131) vs. 16.8% (22/131)] in cancer tissues were higher than those in adjacent tissues (all P<0.001). A total of 131 breast cancer patients were followed up for 3 years without any loss of follow-up, and the follow-up rate was 100%. Within the follow-up for 3 years, there were 15 (11.5%) cases of recurrence, 8 (6.1%) cases of metastasis, and 10 (7.6%) cases of death, the incidence of poor prognosis was 25.2% (33/131); and the remaining 98 cases had good prognosis. The proportions of patients with TNM stage Ⅲ, lymph node metastasis, poorly differentiated histology, tumor diameter ≥3 cm, survivin, p53, and HER2 positive expressions in the poor prognosis group were higher than those in the good prognosis group (all P<0.05). Multivariate logistic regression analysis showed that TNM stage Ⅲ [ OR = 5.323 (95% CI 2.190-12.936)], lymph node metastasis [ OR = 4.773 (95% CI 1.964-11.600)], tumor diameter ≥3 cm [ OR = 3.582(95% CI 1.474-8.706)], positive survivin [ OR = 2.740 (95% CI 1.127-6.659)], positive p53 [ OR = 3.271 (95% CI 1.346-7.949)], and positive HER2 [ OR = 3.873 (95% CI 1.594-9.412)] were independent risk factors for prognosis of breast cancer (all P<0.001). The ROC curve results showed that the area under the curve (AUC) values of survivin positive, p53 positive,HER2 positive, and the combination of any two were more than 0.80 (all P<0.001); the AUC of the combination of the three was 0.944 (95% CI 0.890-0.977) ( P<0.001). Conclusions:The expressions of survivin, p53, and HER2 are highly expressed in breast cancer tissues. The expressions of the three can be used to judge the prognosis of breast cancer patients, and the combination of the three has a higher judgement value.

Objective To investigate the effects of global registry of acute coronary events (GRACE) and Can Rapid risk stratification of unstable angina patients suppress adverse outcomes with early implementation of the ACC/AHA guidelines(CRUSADE) score in the nursing care for patients with acute coronary syndrome (ACS).Methods A total of 128 patients with ACS undergoing percutaneous coronary intervention (PCI) who were admitted from March 2015 to May 2016 were enrolled in this study and divided into usual care group (63 cases) and optimized care group (65 cases). Patients in the usual care group received usual care according to the guidelines, while patients in the optimized care group received optimized care according to GRACE and CRUSADE score. The major adverse cardiac events, the incidence of bleeding complications, average hospitalization and hospitalization costs were recorded and compared between the two groups. Results The characteristics of angiography (multiple vessel rate and target vessel distribution) and procedures of PCI (position, number, length and diameter of stents, coronary blood flow classification after the implantation of stents) were similar (all P>0.05). There were tendencies of reduction in the incidence of MACE, bleeding complications and average hospitalization time in patients of optimized care group compared to the usual care group, with no statistically significant differences between the two groups (P>0.05). The hospitalization costs in the optimized care group (33491±1982)RMB were significantly less than those in the usual care group (36562±2395)RMB. The difference was statistically significant (t=-7.914,P<0.001).Conclusions Optimized care according to GRACE and CRUSADE can improve the treatment effects of ACS patients undergoing PCI and decrease the cost.

Objective:To explore the therapeutic effect of multi-mode sequential combination of artificial liver in the treatment of hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF).Methods:The clinical data of HBV-ACLF patients treated with artificial liver in Wuxi Fifth People′s Hospital from January 2018 to June 2021 were retrospectively analyzed. Eighty-six patients were divided into artificial liver multi-mode sequential combination therapy group (sequential combination group) and conventional treatment group. The cytokine level changes and model for end-stage liver disease (MELD) score were analyzed at 14 days of disease duration. The survival outcome and complications of artificial liver were analyzed after 30 days of follow-up. Two independent samples t test and chi-square test were used for statistical analysis. Cox regression analysis was used to analyze the risk factors of death, and Kaplan-Meier method was used to analyze the survival rate of patients. Results:A total of 86 patients were enrolled, including 48 patients in sequential combination group with the average number of artificial liver of 4.68 times/person, and 38 patients in conventional treatment group with the average number of artificial liver of 3.17 times/person. At 14 days of disease duration, interleukin (IL)-6, IL-8, interferon γ-inducible protein (IP)-10 level and MELD score in sequential combination group decreased significantly than those in the conventional treatment group ( t=3.80, 3.62, 4.95 and 1.11, respectively, all P<0.050). After 30 days of follow-up, 63 patients survived and 23 patients died. Cox regression analysis showed that baseline international normalized ratio (hazard ratio ( HR)=0.558, 95% confidence interval ( CI) 0.193 to 0.856, P=0.027), baseline antithrombin Ⅲ activity ( HR=0.876, 95% CI 0.824 to 0.932, P<0.001), artificial liver mode ( HR=0.819, 95% CI 0.236 to 0.992, P=0.005), spontaneous peritonitis ( HR=0.170, 95% CI 0.045 to 0.647, P=0.009) and hepatic encephalopathy ( HR=0.004, 95% CI 0.001 to 0.030, P<0.001) were independent influencing factors for 30-day survival outcome. The cumulative survival rate of sequential combination group was higher than that of conventional treatment group, and the difference was statistically significant ( χ2=5.45, P=0.020). There were no significant differences in the proportions of bleeding, deep vein thrombosis, heart rate and blood pressure instability between the two groups ( χ2=0.63, 1.20 and 0.54, respectively, all P>0.050). The platelet decline of patients in sequential combination group was slighter than that in conventional treatment group, and the difference was statistically significant ( t=-4.17, P=0.002). Conclusions:Multi-mode sequential combination therapy of artificial liver could eliminate cytokines and reduce MELD score more effectively in patients with HBV-ACLF, and prolong the survival time of patients and have little effect on platelet count.

Objective To investigate the osteogenic properties of a methacrylated gelatin(GelMA)/bone marrow mesenchymal stem cells(BMSCs)composite hydrogel applied to the skull defect area of rats and to provide an experi-mental basis for the development of bone regeneration biomaterials.Methods This study was approved by the Animal Ethics Committee of Nanjing University.A novel photocurable composite biohydrogel was developed by constructing photoinitiators[lthium phenyl(2,4,6-trimethylbenzoyl)phosphinate,LAP],GelMA,and BMSCs.The surface morphology and elemental composition of the gel were examined using scanning electron microscopy(SEM)and energy-dispersive X-ray spectroscopy(EDX).The compressive strength of the gel was evaluated using an electronic universal testing ma-chine.After in vitro culture for 1,2,and 5 days,the proliferation of the BMSCs in the hydrogels was assessed using a CCK-8 assay,and their survival and morphology were examined through confocal microscopy.A 5 mm critical bone de-ficiency model was generated in a rat skull.The group receiving composite hydrogel treatment was referred to as the Gel-MA/BMSCs group,whereas the untreated group served as the control group.At the 4th and 8th weeks,micro-CT scans were taken to measure the bone defect area and new bone index,while at the 8th week,skull samples from the defect ar-ea were subjected to H&E staining,van Gieson staining,and Goldner staining to evaluate the quality of bone regenera-tion and new bone formation.Results SEM observed that the solidified GelMA showed a 3D spongy gel network with uniform morphology,the porosity of GelMA was 73.41％and the pore size of GelMA was(28.75±7.13)μm.EDX results showed that C and O were evenly distributed in the network macroporous structure of hydrogel.The hydrogel compres-sion strength was 152 kPa.On the 5th day of GelMA/BMSCs culture,the cellular morphology transitioned from oval to spindle shaped under microscopic observation,accompanied by a significant increase in cell proliferation(159.4％,as determined by the CCK-8 assay).At 4 weeks after surgery,a 3D reconstructed micro-CT image revealed a minimal re-duction in bone defect size within the control group and abundant new bone formation in the GelMA/BMSCs group.At 8 weeks after surgery,no significant changes were observed in the control group's bone defect area,with only limited evi-dence of new bone growth;however,substantial healing of skull defects was evident in the GelMA/BMSCs group.Quan-titative analysis at both the 4-and 8-week examinations indicated significant improvements in the new bone volume(BV),new bone volume/total bone volume(BV/TV),bone surface(BS),and bone surface/total bone volume(BS/TV)in the GelMA/BMSCs group compared to those in the control group(P＜0.05).Histological staining showed continuous and dense formation of bone tissue within the defects in the GelMA/BMSCs group and only sporadic formation of new bone,primarily consisting of fibrous connective tissue,at the defect edge in the control group.Conclusion Photocur-ing hydrogel-based stem cell therapy exhibits favorable biosafety profiles and has potential for clinical application by inducing new bone formation and promoting maturation within rat skull defects.

Objective To explore the relationship of high-sensitivity C-reactive protein (Hs-CRP), homocysteine (HCY) levels and the short-term outcome of acute ischemic stroke (AIS). Methods 189 patients with AIS were included. Serum Hs-CRP, HCY levels and National Institute of Health Stroke Scale (NIHSS) were assessed after admission. Short- term functional outcome was measured by modified Rankin Scale (mRS) 90 d after admission. Results The serum Hs-CRP and HCY levels were significantly higher in AIS patients than in normal controls (P<0.001). Adjusting age and the NIHSS score, Hs-CRP and HCY were independent prognostic markers of functional outcome and death in patients with AIS. In receiver operating characteristic curve (ROC) analysis, the prognostic accuracy of combined model (HCY and Hs-CRP) was higher than that of biomarkers measured alone and the NIHSS score. Conclusion Hs-CRP and HCY are independent predictors of short-term outcome and mortality after AIS. Combined model may provide additional general prognostic information.