OBJECTIVE To establish an HPLC method for determination of the related substances in papaverine hydrochloride. METHODS NanoChrom ChromCore 120 C8 column was used; the mobile phase A consisted of 3.4 g·L−1 potassium dihydrogen phosphate aqueous solution, adjust pH to 3.5 with phosphoric acid-acetonitrile(90∶10), the mobile phase B was methanol, with gradient elution at the flow rate of 0.8 mL·min−1; the detection wavelength was 238 nm; the column temperature was 50 ℃. RESULTS The minimum separation between the main component and each impurity was >1.5; Papaverine and its thirteen impurities showed a good linear relationship in the self-concentration range(r>0.999); and the average recoveries were 93.1%−101.2% with RSDs of 2.3%−8.1%. CONCLUSION The method is accurate, sensitive and reliable, which is suitable for the determination of related substances in papaverine hydrochloride.

Objective To investigate the protective effects of Sulforaphane(SFN)against high glucose(HG)-induced human platelet apoptosis and elucidate the underlying mechanisms in vitro.Methods Purified platelets obtained from healthy individuals were pre-incubated with various concentrations of SFN(5,10,or 20 μmol/L)or a vehicle control(0.05%DMSO)for 40 minutes at 37℃.Subsequently,the platelets were stimulated with normal glucose(NG,5 mmol/L)or high glucose(HG,25 mmol/L)for an additional duration of 90 minutes.Flow cytometry was employed to assess platelet mitochondrial membrane potential depolarization(Δψm),exposure of phosphatidylserine(PS),and generation of total intraplatelet reactive oxygen species(ROS).Phosphorylation levels of PI3K and Akt were determined using Western blot.Results Compared to NG-treated human platelets,HG significantly induced depolarization of platelet Δψm and exposure of PS(P＜0.001).These effects of HG were markedly attenuated by various concentrations of SFN(P＜0.001).Mechanistically,SFN down-regulated the phosphorylation levels of PI3K(P＜0.01)and Akt(P＜0.05),which were increased by HG when compared to the vehicle control,and substantially reduced total intracellular ROS levels(P＜0.001).The inhibitory effects of SFN on HG-induced phosphorylation of PI3K and Akt,as well as its efficacy on total intracellular ROS generation,Δψm depolarization,and PS exposure in HG-stimulated human platelets were completely reversed by a specific agonist for PI3K,740 Y-P.Conclusions The current study demonstrates that SFN exerts a protective effect on platelet apoptosis induced by HG,potentially through the down-regulation of the PI3K/Akt-mediated pathway in human platelets in vitro.These findings suggest that SFN may hold promise for improving thrombosis in diabetes mellitus and related chronic metabolic diseases.

Objective To investigate the role of Taraxasterol(TAR)on ferroptosis in chondrocytes induced by Erastin.Methods The C28/I2 chondrocyte line was treated with Erastin to construct the ferroptosis model of chon-drocytes in vitro and the experiments were divided into Control,Erastin,TAR,and TAR+Erastin groups.Cell via-bility was detected by the CCK-8 assay.Cytotoxicity was detected by the lactate dehydrogenase(LDH)kit and the Calcein/PI cytokinesis kit.Flow cytometry was used to detect lipid reactive oxygen species(ROS).The intracellular glutathione(GSH)content was detected by GSH kit.Mitochondrial membrane potential was detected by JC-1 stai-ning and RH123 staining.ACSL4 and GPX4 protein expression and the key indicators of ferroptosis were detected by Western blot.Results TAR restored the decreased cell viability of C28/I2 chondrocytes induced by Erastin treatment as well as reduced Erastin-induced cytotoxicity(P＜0.01).Compared with the control group,the level of intracellular lipid ROS increased(P＜0.01)and the content of GSH decreased(P＜0.01)after treatment with Erastin,while TAR could reduce the production of lipid ROS(P＜0.01)and increase the content of GSH(P＜0.01).TAR restored mitochondrial membrane potential in C28/I2 chondrocytes ferroptosis,decreased ACSL4 pro-tein expression(P＜0.01)and increased GPX4 protein expression(P＜0.01).In addition,TAR restored the re-duced cell viability caused by IL-1 β treatment.Conclusion TAR can inhibit Erastin induced ferroptosis in C28/I2 chondrocytes,which may be related to the regulation of ACSL4 and GPX4 protein expression.

Objective To investigate the role and possible mechanism of curcumin(Cur)regulating Peroxiredoxin-6(Prdx6)expression in inhibiting Erastin-induced ferroptosis in C28/I2 chondrocytes.Methods Safranin O/Fast Green and hematoxylin-eosin(HE)staining were performed to observe the pathological changes in the knee joint of rats with osteoarthritis(OA).The expression levels of Prdx6 and GPX4 proteins in cartilage tissues with OA were detected by immunohistochemistry and Western blot.C28/I2 chondrocytes were treated with different concentrations of Cur,cell viability was detected by thiazolyl blue tetrazolium bromide(MTT)assay and cytotoxicity was measured by lactic dehydrogenase(LDH)assay.The production of lipid reactive oxygen species(ROS)in chondrocytes was detected by flow cytometry,and the total glutathione(GSH)assay kit was used to detect the GSH level in chondro-cytes.Western blot was performed to detect the expression level of Prdx6 and ferroptosis-related proteins in chon-drocytes.The interaction between the Cur molecule and Prdx6 was analyzed through the molecular docking tech-nique.Results During the OA progression,OA rats and OA patients showed pathological changes such as damage to the cartilage and a decrease in the number of chondrocytes.The expression levels of Prdx6 and GPX4 were re-duced in the cartilage tissues of OA patients compared with healthy people.Further study revealed that the treat-ment of Erastin-induced ferroptosis in C28/I2 chondrocytes in a mouse model with 20 μmol/L of Cur could improve cell viability,decrease cytotoxicity,inhibit lipid ROS production,and increase the level of intracellular GSH.Western blot results showed decreased expression of Prdx6,SLC7A11,FTH,and GPX4 and increased expression of ACSL4.In addition,Cur molecules interacted with Prdx6 protein by van der Waals forces and π bond.Conclu-sion Cur may inhibit Erastin-induced ferroptosis in C28/I2 chondrocytes by upregulating the Prdx6 expression lev-el.

Objective:To explore the effects of operation timing on postoperative complications and operative duration in children undergoing non-emergency operation for femoral neck fracture.Methods:Fifty-nine children and adolescents with femoral neck fracture were retrospectively analyzed who had been admitted to Department of Pediatric Orthopedics, Zhengzhou Orthopedic Hospital Affiliated to Henan University from February, 2015 to September, 2019. They were 39 boys and 20 girls with a mean age of 11.7 (7.1, 13.7) years. By Delbet fracture classification, 7 cases were type Ⅰ, 27 cases type Ⅱ, 20 cases type Ⅲ, and 5 cases type Ⅳ. The patients were divided into 4 groups by the time from injury to operation (TFITO): 17 cases were assigned into group A with TFITO from 24 to 48 hours, 14 cases into group B with TFITO from 49 to 72 hours, 12 cases into group C with TFITO from 73 to 96 hours, and 16 cases into group D with TFITO>96 hours. The effects of TFITO on postoperative complications and operative duration were analyzed.Results:There were no significant differences between the 4 groups in their preoperative general data, showing they were comparable ( P>0.05). This cohort was followed up for 12 to 62 months (average, 20 months). The operation time was not included for this study in 6 cases whose associated injury had to be treated simultaneously. The median operation time for the other 53 patients was 80 (70, 105) min. The correlation coefficient between TFITO and operation time was 0.098 ( P=0.484). Postoperative complications occurred in 37.3% of the patients (22/59), including 14 cases of avascular necrosis of femoral head. For groups A, B, C and D, the incidences of complications were 47.1% (8/17), 50.0% (7/14), 25.0% (3/12) and 25.0% (4/16) while the incidences of avascular necrosis of femoral head 23.5% (4/17), 31.3% (5/16), 16.7% (2/12) and 18.8% (3/16), showing insignificant differences between the 4 groups in all the comparisons ( P>0.05). Conclusion:The time from injury to operation may not increase operative duration or postoperative complications such as avascular necrosis of femoral head in children undergoing non-emergency operation for femoral neck fracture.

BACKGROUND: Preliminary study has prepared the three-dimensional silk fibroin/chitosan/nano-hydroxyapatite scaffold successfully.OBJECTIVE: To explore the mechanical properties, physical characteristics, chemical composition and antibiotic sustained-release ability of three-dimensional silk fibroin/chitosan/nano-hydroxyapatite scaffold loaded with levofloxacin. METHODS: Levofloxacin/chitosan (3:1) microspheres were constructed by emulsion settlement filter method. 5, 7.5 and 10 g of microspheres were added into 2％ of silk fibroin/chitosan/nano-hydroxyapatite mixed solution through freeze drying and chemical cross-linking to obtain the scaffolds loaded with antibiotics. The scaffolds loaded with antibiotics underwent scanning electron microscope observation, and chemical composition analysis. The sustained release, mechanical properties, porosity, water absorption expansion rate and hot water soluble loss rate were detected. RESULTS AND CONCLUSION: (1) Scanning electron microscope observed that there were drug microspheres at the inner wall of the scaffold, and the voidage was decreased with mass of microspheres increasing. (2) Energy spectrum analysis showed that the three kinds of scaffolds were rich in calcium and phosphonium ions. (3) The three kinds of scaffolds showed the same releasing trend, which presented with sudden-release effect at the former 3 days (release> 50％) , and then tended to be stable. The release rate was the slowest in the scaffold loaded with 10 g of microscopes, and the rapidest in the scaffold loaded with 5 g of microscopes. (4) With the mass of microspheres increasing, there was an increase in the compressive and tension abilities and hot water soluble loss rate, and a decrease in the porosity, mean pore size and water absorption expansion rate. (5) These results indicate that the three-dimensional tissue-engineered scaffold loaded with levofloxacin is constructed successfully by freeze drying and chemical cross-linking method, which holds good sustained-release effect and compressive ability, water absorption expansion rate and hot water soluble loss rate.

Objective To evaluate the efficacy of volume therapy guided by stroke volume variabil-ity ( SVV) in the patients undergoing surgery for severe traumatic brain injury. Methods Thirty patients of both sexes with severe traumatic brain injury, aged 18-64 yr, of American Society of Anesthesiologists physical status Ⅲ, who were admitted to the hospital within 24 h after injury, with Glasgow Coma Scale ( GCS) score≤8, were divided into control group ( C group, n=15) and SVV group ( n=15) using a ran-dom number table method. In group C, conventional fluid administration was performed to maintain mean arterial pressure at 65-110 mmHg, central venous pressure at 5-12 cmH2 O and urine volume>1 ml·kg-1 ·h-1 . Fluid was given according to SVV, maintaining SVV≤13％ and mean arterial pressure at 65-110 mmHg in group SVV. Immediately after skin incision ( T0 ) , immediately after opening cerebral dura mater ( T1 ) , at 1 h after opening cerebral dura mater ( T2 ) , immediately after suturing cerebral dura mater ( T3 ) and at the end of operation ( T4 ) , blood samples were collected from the radial artery and inter-nal jugular venous bulb for blood gas analysis, the jugular venous oxygen partial pressure, jugular venous bulb oxygen saturation, blood lactate, arterial oxygen partial pressure, arterial oxygen saturation and Hbwere recorded, and the cerebral artery and arteriovenous blood O2 content difference and cerebral O2 extrac-tion rate were calculated. Blood samples were collected from the internal jugular venous bulb at T0-2 , T4 and 24 h after operation ( T5 ) for determination of S100β protein concentrations by enzyme-linked immunosor-bent assay. The intraoperative volume of fluid intake and output and consumption of vasoactive drugs were recorded. GCS scores were recorded immediately after admission to the operating room, and at 1, 3, 7 and 14 days after operation. The development of postoperative length of hospitalization and complications ( pul-monary infection and brain edema) was recorded. Glasgow Outcome Scale Score was used to assess the early postoperative quality of life. Results Compared with group C, the urine volume was significantly in-creased, the consumption of vasoactive drugs was reduced, jugular venous bulb oxygen saturation was in-creased at T2,3 , the cerebral O2 extraction rate was decreased at T2-4 , the serum S100β protein concentra-tion was decreased at T2 , and the GCS score was increased at day 3 after operation ( P<0. 05) , and no sig-nificant change was found in blood lactate, postoperative Glasgow Outcome Scale score or length of hospital-ization at each time point in group SVV ( P>0. 05) . Conclusion SVV-guided volume therapy can improve cerebral oxygen metabolism, ensure adequate tissue perfusion and reduce craniocerebral injury in the pa-tients undergoing surgery for severe traumatic brain injury.

Objective To explore the factors imfluencing the prognosis of patients with renal cell carcinoma after laparoscopic partial nephrectomy.Methods Clinical data from 593 renal cell carcinoma patients underwent laparoscopic partial nephrectomy in our institution from December 2010 to December 2015 were retrospectively collected..Tbere were 396 males and 197 females,aged 35 to 72 years old(mean 55.4 years).There were 181 cases of smoking history,206 cases of hypertension and 105 cases of diabetes.Before operation,98 cases of liver function were damaged and 122 cases were anemia.There were 521 cases with PLT ＜ 450 × 109/L,72 cases PLT≥ 450 × 109/L.Thc tumor of 292 cases on the left and 301 cases on the right.The Kaplan-Meier method log-rank test was used for survival analysis.Univariate analysis and factors which were significantly associated with survival in the univariate analysis were conducted into the multivariate analysis with Cox proportional hazards model.Results The operative time was 88.0-120.6 min,mean 104.3 min,the blood loss during operation was 47.2-157.8 ml,mean 102.5 ml. Pathological tumor stage revealed that 398 cases were T1 and 195 cases were T2 . Fuhrman classification revealed that 29 cases were grade Ⅰ,411 cases were grade Ⅱ,150 cases grade Ⅲ,3 cases grade Ⅳ . The median follow-up time was 36 months(ranged 6 to 99 months) . Five cases (1.3％) in the T1 patients relapsed,recurrencefree survival (RFS) rate was 98.7％;In the T2 patients,7 cases (3.6％) relapsed,1 of them had local recurrence and died of lung metastasis,and the RFS rate was 96.4％.Univariate analysis revealed that T stage,Fuhrman grade,tunor necrosis,tumor pseudocapsule,lymphovascular invasion,collection system violation,anemia,and high platelet were significantly associated with RFS of patients.Multivariate analysis found that T stage (HR =1.524,95 ％ CI 1.326-1.926,P =0.001),Fuhrman grade (HR =1.600,95 ％ CI 1.035-2.364,P =0.022),tumor necrosis (HR =2.315,95％ CI 1.523-3.624,P =0.001) were independent risk factors for the RFS of patients with renal cell carcinoma after retroperitoneal laparoscopic partial nephrectomy.Conclusion High T stage,high Fuhrman grade and tumor necrosis were independent risk factors for the RFS of patients with renal cell carcinoma after retroperitoneal laparoscopic partial nephrectomy.

Objective To provide the anatomical basis for the flap based on the perforator of Plantar arch,through investigation of the morphological features of the perforator of the arch of the foot.Methods From November,2015 to March,2016,the first metatarsal base and the fifth metatarsal tuberosity were chosen as the observation point on 25 specimens of adult human feet perfused with red latex.The following contents were observed under surgical magnifier:①The origin,courses,branches and distribution of the perforator of Plantar arch.②The anastomoses among the perforator of Plantar arch and the fete arteriosum dorsale pedis.Mimic operation was performed on another fresh specimen perfused with red latex.Results There were 3 perforators in Plantar arch,which passed through the 2nd-4th metatarsal dorsal muscles to the dorsi pedis and then divided into an ascending branch and a descending branch.The ascending branch anastomosed with the rete arteriosum dorsale pedis,and the descending branch stretched to the 2nd-4th plantar arteries.The initiative outer diameters of the 1st-3rd dorsal perforators of Plantar arch were (1.5 ± 0.3)mm,(1.1 ± 0.4) mm and (0.9-± 0.3) mm respectively,and the lengths of the stem were (1.1 ± 0.2) cm,(1.5 ± 0.1) cm and (1.5 ± 0.5) cm respectively.Conclusion The flap can be used for repair of soft-tissue defects of dorsal and front foot through dorsal transposition or a V-Y advancing flap with the perforator of Plantar arch as its vascular pedicle.

Objective To investigate the efficacy of percutaneous transhepatic variceal embolization (PTVE) with Cyanoacrylate (TH glue) in treating large gastric fundal variceal bleeding.Methods PTVE was performed on 24 patients with TH glue injected into the main stem of left gastric vein and its fundic branches.The degree of varices in gastric fundus,rebleeding rate and survival rate after the procedure were compared with those before.Results Varices in gastric fundus were all embolized successfully with TH glue.The diameter of varices was reduced to below 5mm or disappeared in 20 patients (83.3％),and reduced to 5-10mm in the other 4 ( 16.7％ ) During the follow-up period of 3-36 months(mean 16.6 months),the rebleeding rate and mortality were 12.5 ％ ( 3/24 ),and 12.5 ％ (3/24),respectively.One patient died of liver cancer,and two others died of chronic liver failure.Conclusion PTVE with TH glue is of ideal therapeutic effect to block the feeding veins of large gastric fundal varices.