[This corrects the article DOI: 10.1016/j.apsb.2022.07.023.].

As a promising modality for cancer therapy, photodynamic therapy (PDT) still acquired limited success in clinical nowadays due to the extremely serious hypoxia and immunosuppression tumor microenvironment. To ameliorate such a situation, we rationally designed and prepared cascade two-stage re-oxygenation and immune re-sensitization BSA-MHI148@SRF nanoparticles via hydrophilic and hydrophobic self-assembly strategy by using near-infrared photodynamic dye MHI148 chemically modified bovine serum albumin (BSA-MHI148) and multi-kinase inhibitor Sorafenib (SRF) as a novel tumor oxygen and immune microenvironment regulation drug. Benefiting from the accumulation of SRF in tumors, BSA-MHI148@SRF nanoparticles dramatically enhanced the PDT efficacy by promoting cascade two-stage tumor re-oxygenation mechanisms: (i) SRF decreased tumor oxygen consumption via inhibiting mitochondria respiratory. (ii) SRF increased the oxygen supply via inducing tumor vessel normalization. Meanwhile, the immunosuppression micro-environment was also obviously reversed by two-stage immune re-sensitization as follows: (i) Enhanced immunogenic cell death (ICD) production amplified by BSA-MHI148@SRF induced reactive oxygen species (ROS) generation enhanced T cell infiltration and improve its tumor cell killing ability. (ii) BSA-MHI148@SRF amplified tumor vessel normalization by VEGF inhibition also obviously reversed the tumor immune-suppression microenvironment. Finally, the growth of solid tumors was significantly depressed by such well-designed BSA-MHI148@SRF nanoparticles, which could be potential for clinical cancer therapy.

Objective To evaluate the clinical value of adriamycin injection via the foramen ovale and around peripheral trigeminal branches under the guidance of X-ray for treatment of primary trigeminal neuralgia by comparison with the three-dimensional computed tomography (CT).Methods A total of 91 patients with primary trigeminal neuralgia of both sexes,aged 33-76 yr,with the course of disease 6 months-24 yr,with visual analogue scale score of 6-9,were divided into 2 groups using a random number table:X-ray group (n =43) and CT group (n =48).Hartel anterior approach was used to puncture the foramen ovale in 2 groups.One point five percent adriamycin 0.2,0.3 and 0.5 ml were injected via the supraorbital foramen,infraorbital foramen and oval foramen.When pain relief was poor (visual analogue scalc scorc≥ 4) within 1 yr after treatment,oxcarbazepine and adjuncts (tramadol,flupentixol and melitracen tablets,etc.) were taken orally.The requirement for oxcarbazepine and adjuncts was recorded during 1 day-1 week,1 week-1 month,1-3 months,3-6 months and 6 months-1 yr after treatment periods.The operation time,the nuinber of puncture,and developinent and recurrence of complications during treatment and within 1 yr after treatment were recorded.Results Compared with CT group,the number of puncture and incidence of facial hematoma during treatment were significantly increased (P ＜ 0.05 or 0.01),and no significant change was found in the operation time,requirement for oxcarbazepine and adjuncts,incidence of dizziness,nausea and vomiting during treatment,or the incidence and recurrence rate of masticatory muscle weakness and facial numbness after treatment in X-ray group (P＞0.05).Conclusion Compared with the three dimensional CT,X-ray provides similar efficacy and safety when used to guide adriamycin injection via the foramen ovale and around peripheral trigeminal branches for treatment of primary trigeminal neuralgia,showing that X-ray guidance has significant clinical value.

Objective To discuss the diagnostic value of peritoneal effusion detection by emergency-ultrasound in patients with closed abdominal injury. Method From August 2006 to June 2009,212 patients with closed abdominal injury were studied to evaluate peritoneal effusion detection by emergency ultrasound. Results of 212 patients,peritoneal effusion frequency rate was 78.8%( 167/212), meanwhile,abdominal paracentesis confirmation ratio was only 46.2%(98/212). In the follow-up, 13 patients with injuried hollow viscera and 1 patient with rupture of kidney showed peritoneal effusion. The volume of abdominal fluid was increasing in 17 patients,which needed to be managed by surgery. The accuracy rates were respectively 78.3%( 112/143) and 36.1%(13/36) in the solid organs and the hollow organs. Conclusion During the course of diagnosis and treatment in closed abdominal injury,peritoneal effusion monitoring by ultrasound should be used routinely, which can help to decrease the rate of misdiagnosis and avoid delayed treatment.

Objective To compare the clinical efficacy and safety of isotretinoin erythromycin gel, a gel containing isotretinoin (0.05%) and erythromycin (2%), versus adapalene gel in the treatment of mild to moderate acne vulgaris. Methods A multicenter, randomized, open, parallel-controlled clinical study was conducted. A total of 192 patients with mild to moderate (Grade Ⅰ -Ⅲ ) acne vulgaris were enrolled in this study according to the grading criteria for acne severity in guidelines for the treatment of acne in China. Efficacy analysis was carried out in 169 patients and safety analysis in 190 patients. The patients were classified into trial group (n = 86) and control group (n = 83 ) to be treated with isotretinoin erythromycin gel or adapalene gel once a night for 6 weeks. Patients were evaluated at the baseline, on week 2, 4 and 6 during the treatment for the count of comedones (both open and closed), inflammatory papules and pustules, severity of acne and local or general adverse effects. Results After the start of treatment, the response rate gradually increased and severity of acne decreased in both groups. On week 6, the total response rate was 51.16% in the trial group and 40.96% in the control group (P ＞ 0.05), while a greater reduction in the count of pustules and inflammatory lesions was observed on week 4 and 6 in the trial group with a lower severity grade of acne compared with the control group (P ＜ 0.05 or 0.01 ). Adverse reactions were similar in both groups and manifested as tolerable local irritation. Conclusions The efficacy of isotretinoin erythromycin gel is similar to that of adapalene gel in the treatment of mild to moderate acne vulgaris, however, isotretinoin erythromycin gel seems superior to adapalene gel in reducing inflammatory lesions and rapidly improving severity of acne vulgaris.

BACKGROUND:Mesenchymal stem cells (MSCs) exist in human tissues.Presently,cell source is single;culture method has great differences;obtained results are not consistent.Thus,it cannot verfy that isolated and cultured cells are identical calls,which is difficult to compare.OBJECTIVE:To compare the biological features of MSCs derived form bone marrow (BM),perpheral blood (PB) and cord blood (CB) under in vitro culture conditions.DESIGN,TIME AND SETTING:The cytological in vitro controlled study was performed at the Department of Hematology,Navy General Hospital of Chinese PLA from June 2007 to December 2008.MATERIALS:A total of 10 donors of hemopoietic stem cell transplantation at the Department of Hematology,Navy General Hospital of Chinese PLA were selected.MB and PB cells were obtained from the same donor,and cell volumes were respectively 20 mL and 2 mL.CB cells (30 mL) were obtained from healthy primipara at the Department of Obstetrics,Navy General Hospital of Chinese PLA.METHODS:MSCs were obtained from BM,PB and CB by Percoll density gradient + adherence method,and then incubated in DMEM/F12 medium containing 10% fetal bovine serum.When 80%-90% confluency,cells were digested in trypsin-EDTA and made into 5×10~8/L cell suspension as P_0.Above-described operation was performed as P_1,and the rest may be deduced by analogy as P_2-P_5.MAIN OUTCOME MEASURES:The following parameters were measured:cell growth morphology;results of Wright-Giemsa staining;results of cytochemistry;cell proliferation amount;cell surface markers using flow cytometry.RESULTS:Time of adherence,time to 50% confluency and time to 80% confluency of BMSCs were earlier comarped with the PBMSCs and UCMSCs.Adherent cells from BM grew in whirpool-like type,while CB and PB did not at 5-7 days.Majority of aderent cells from BM were fibroblast-like cells,and small parts were endothelioid cells.Aderent cells from PB and CB at the fifth generation contained more endothelioid cells and mononuclear and macrophage-like cells besides fibroblast-like cells.PAS stain,Sudan black B stein,alkaline phosphatase (AKP) staining of adherent cells from BM,PB and CB were negative from P_1 to P_5.Compared with P0 cells,number of BMMSCs till P5 was significantly more in PBMSCs and UCMSCs (P ＜ 0.05).Positive rates of CD29,CD44,CD90,CD71,CD105,CD166 and HLA-ABC were 55.9% 92.8% at P0 to P5,but ≤6% following BMMSCs were incubated;19.7%-33.4% at P0 to P5,but ≤10% following PBMSCs were incubated;35.4%-93.2% at P_0 to P_5,but ≤20% following CBMSCs were incubated.Positive rates of CD34,CD45 and HLA-DR were low in BM-,PB-and CB-MSCs.Positive rates of CD14 and CD31 were low in BMMSCs;12.1%-28.3% in PBMSCs,and 8.1%-21.3% in CBMSCs.CONCLUSION:MSCs can be attained from BM,PB and CB.Quantities of MSCs form BM are the highest,with single component,followed by CBMSCs and PBMSCs,with multiple components.

BACKGROUND: It is important to study the methods of culturing bone marrow-derived mesenchymal stem cells (MSCs) to obtain a great amount of high purity MSCs for applying ocular tissues constructed by tissue engineering technique to treat eye diseases.OBJECTTVE: To separate and culture in vitro MSCs from bone marrow of the adult rats by density gradient centrifugation combined with adherence culture, and observe the growing characteristics and the possibility of mass multiplication.DESIGN: A completely randomized grouping design/repetitive measuring experiment.SETTING: Pathological laboratory, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: Four six-week-old SD rats about 250 g, grade Ⅱ of cleaning, were provided by the Animal Center of Sun Yat-sen University [certificate number: SCSK(Yue)2004/0011], about 250 g each rat and there was no limit to the sex. The main reagents and instruments included low sugar Dulbecco modified Eagle culture medium (DMEM/F12, American Gibco Corporation), trypsin (fetal bovine serum (FBS, Hangzhou Sijiqing Bio-Engineering Material Research Institute), American Gibco Corporation), disodium edetate, lymphocyte separating medium, fibronectin, CD44, CD34, CD31 monoclonal antibodies, two-step-method kit for immunohistochemistry (Beijing Zhong shan Biotechnology Corporation).METHODS: This experiment was conducted at the Key laboratory of Ophthalmology (Sun Yat-sen University), Ministry of ethanol (750 g/L) for 10 minutes. Under aseptic condition, the medullaris cavitas was exposed, the syringe containing application m edium was directly punctured into the femoral cavity, the cells in the medullaris cavitas were washed out with the culture medium containing heparin and taken as the cell suspension. The bone marrow-derived MSCs were separated and purified by density gradient centrifugation combined with adherence culture, and the growing conditions of the wells. When the cells had generally connected with each other, they were fixed with methanol or dimethy ketone in situ for 10 minutes, and then hematoxylin eosin (HE) staining or immunohistochemical staining. Antibodies against fiinoculated to 96-well culture plate by a cell density of 4.25×107/L, with 200 μL every well; then put the culture plate into culture box. Then from the next day to the sixth day, 5 g/L MTT solution was added into two rows (20 μL every well) every day, continuously cultured for 4 hours, then the supernatant was removed, and 200 μL DMSO was added to each well, agitated for 5 minutes, then detected the absorbance (A) values at 570 nm wave length, and a growth curve was drawn.branes were clear and the cell bodies were lucent. Being cultured for 2 days, there appeared adherent cells. 3 days later,most of the adherent cells extended and appeared to be in polygon, star or long-shuttle shapes. 4 days later, the cells showed to be in division growth stage; and about 12 days later, the cell clones were connected to each other, appearing curve of subcultured MSCs was in S shape. Cells began growing fast on the 2nd day after being passaged, and they entered the growth period for 3-4 days followed by saturation period, and then cells stopped growing.CONCLUSION: It is a simple and practical method to separate MSCs from the bone marrow of adult rats by means of density gradient centrifugation and adherence. MSCs can greatly proliferate in vitro and offer seed cells for the application of tissue engineering technique to treat eye diseases.

Objective:To investigate the effects of the β-casomorphin-7 on in vitro and in vivo mice splenic lymphocyte and macrophage nitric oxide production.Methods:Splenocytes proliferation assay and nitrite determination were employed for the studies of effects of β-casomorphin-7 on mice splenic lymphocyte and macrophage following in vitro stimulation of β-casomorphin-7 and in vivo intrapetitoneal administration and drinking β-casomorphin-7 solution administration.Results:In vitro study, β-casomorphin-7 showed a stimulation as well as a suppression of lymphocyte proliferation and significantly(P＜0.01) suppressed the production of nitric oxide. In vivo study, β-casomorphin-7 actions on lymphocytes and macrophages were accordant in intraperitoneal administration and drinking β-casomorphin-7 solution administration. β-casomorphin-7 significantly(P＜0.01) increased in splenic lymphocyte proliferative response and suppressed nitric oxide production in peritoneal macrophages.Conclusion:The present study indicates that the β-casomorphin-7 has the immunomodulatory effects and β-casomorphin-7 may be permeable into peripheral blood intact in mice of 2-3 weeks of age.

In order to estimate the relationship between iron and the Alzheimer's disease, the behavioral and pathological changes were observed by Morris water maze and immunohistochemical staining respectively after injecting FeC13 into brain ventricle of rats. The apoptosis was tested by flow cytometry and the electron microscopy was used to observe ultrastructural changes. There were significant differences in escape latency of time and distance between normal animals and iron treated rats. Percentage and fluorescence intension (FI) of AnnexinV FITC loaded cells undergoing apoptosis were higher in iron treated rats compared with normal animals. Fawn-coloured products of β amyloid protein were interspersedly distributed in extensive areas of cerebral cortex and hippocampus. Under electron microscope, vacuolate degeneration of neuronal processes with mitochondria degeneration and accumulation of microtubule near vacuolar nucleus were observed in iron treated rats. These results suggest that a local higher concentration of iron in brain may induce Alzheimer-like impairment of intelligence and pathological changes.

·AIM: To investigate the preparation of endostatin protein and its biologic activity on vascular endothelial cell.· METHODS: pBlast-hEndostatin and pBlast-Mcs were identified by digesting with Nhe Ⅰ and Sal Ⅰ, by PCR reaction, by sequencing, and by Alignments of PCR products with gene bank using NCBIBLAST software. The identified pBlast-hEndostatin as well as pBlast-Mcs were then purified with QIAGEN Endofree plasmid maxi kit.The purified plasmids transfected human fibroblasts. The expression of endostatin was detected by RT-PCR, Westem-Blot and immunohistochemistry. The endostatin prorein produced by transfected fibroblasts was purified by ultrafiltration and affinity chromatography. The inhibitory action of endostatin on human umbilical vein endothelium was measured by MTT assay.· RESULTS: pBlast-hEndostatin was found to contain human endostatin gene. Endostatin protein was produced by transfected fibroblasts. The inhibitory ratio of 2.5,5,10,20,40,80mg/L endostatin on human umbilical vein endothelium for 48h were 8.5%,13.1%,27.7%,38.1%,56.7%,63.8% respectively. IC50 value was 34.5mg/L.No inhibition action was found on fibroblasts.·CONCLUSIONS: Endostatin protein can be produced by the transfected fibroblasts. The produced endostatin has inhibitory action on human umbilical vein endothelium and has no inhibition action on fibroblasts.