Objective To study the estradiolNGF regulatory cascade in retinal endothelial cells. Methods Retinachoroids vascular endothelial cell line RF/6A from rhesus was cultured in M199 medium contain 20% FBS.mRNA of ER,NGF and its receptors,TrkA and P75 NTR were detected with RTPCR.The cells were incubated with NGF,VEGF,antibodies against NGF or VEGF,and K252a(100nmol/L),the specific inhibitor of TrkA,separately or in different combination.MTT based cell counting assay was used to study the viability of the cells.The apoptosis was evaluated by FACS,mass migration by wound healing assay,and tubogenesis by AngioMatrix assay. Results We amplified the specific fragments of cDNA of ER,NGF and NGF receptor TrkA using RTPCR.10?nmol/L1??mol/L estradiol augments the proliferation and increases the viability of RF/6A in a dosage dependent manner.In the wound healing based migration assay,we found the similar alteration.This effects of estradiol was partially blocked by NGF neutralized antibody and K252a.Apoptosis rates were at the similar level among the groups.For the tubogenesis of RF/6A,we found no augmentation by NGF,and no blocked augmentation by estradiol.Conclusion NGF,first identified as the survival factor of nerve system,now also seemed to be an activator of retinal endothelial cell,is under the regulation of estrogen.The proliferation and migration,but not the tubogenesis of retinal endothelial cells are regulated by this regulatory cascade.

OBJECTIVETo observe the therapeutic efficacy of dietary boron supplement on retinoic acid-induced osteoporosis in rats, so as to provide experimental evidence for clinical management of osteoporosis with boron.

METHODSThirty-two SD rats were randomized into normal control group (8 rats) and osteoporotic group (24 rats), and osteoporosis was induced in rats of the latter group by intragastric retinoic acid administration at the daily dose of 80 mg/kg for 15 consecutive days. The osteoporotic rats were subdivided into control group (8 rats) without treatment, boron treatment group (8 rats) and estradiol treatment group (8 rats). After 30 days of treatment, the serum contents of Ca, P, boron and the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) in the rats were assayed, the bone mineral density (BMD) of the whole body, lumbar vertebrae and tibia were determined, and the morphological changes of the femurs were observed.

RESULTSThe serum contents of Ca and P in the rats of the 4 groups differed scarcely, but the content of boron in boron treatment group was markedly higher than that in the other three groups. In the osteoporotic control group, the activities of serum AKP and TRAP, the masses of spongy bone and cortical bone of the femurs, and the quantity of the osteoclasts were increased, with the BMD of the lumbar vertebrae and tibia decreased, suggesting osteoporotic conditions. The mean trabecular plate density and thickness, trabecular bone volume and cortical bone volume of the femurs in the osteoporotic rats treated with boron or estradiol were significantly increased, but the active osteoclast quantity in the spongy bone and serum TRAP activities were obviously decreased, and the bone quality was comparable with that of the normal group. In addition, the serum AKP activity and the active osteoblast quantity in the spongy bone were obviously increased in boron treatment group.

CONCLUSIONThe dietary boron supplement can increase the serum content of boron of osteoporotic rats to stimulate bone formation and inhibit bone resorption, producing therefore obvious therapeutical effect against osteoporosis.

Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Bone Density ; drug effects ; Boron ; administration & dosage ; therapeutic use ; Dietary Supplements ; Female ; Femur ; drug effects ; growth & development ; metabolism ; Isoenzymes ; blood ; Osteoporosis ; blood ; chemically induced ; drug therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase ; Time Factors ; Tretinoin

Objective To explore the effect of down regulation of EZH2 expression by shRNA on apoptosis and proliferation in colorectal cancer cell.Methods The human colorectal cancer cell line HCT116 was used as the materials.The EZH2 expression was knockdown by traninfecting siRNA.The EZH2 protein expression was tested by western-blot array, the HCT116 apoptosis rate was tested by flow cytometry array and the HCT116 cell proliferation ability was tested by MTT array.Results The EZH2 protein significant decreased by si -EZH2 ( P <0.05 ) .The apoptosis rate significant increased by knockdown the EZH2 expression ( P<0.05) .And the proliferation ability was inhibited after knockdown the EZH2 expression ( P<0.05).Conclusion EZH2 may be involved in the process of apoptosis and proliferation in human colorectal cancer cells which could be a potential target for treatment.

OBJECTIVETo study clinical effects of PHILOS (proximal humeral internal locking system) plates through mini-open deltoid-splitting approach for the treatment of proximal humeral fractures.

METHODSFrom March 2006 to August 2010, 22 patients with proximal humeral fractures were treated with PHILOS plates through mini-open deltoid-splitting approach. According to Neer classification, 6 cases were type II, 15 cases were type III and 1 case was type IV. Through the anterolateral approach to the shoulder, anterolateral vertical incision of 4 cm length was perforrmed from 1 cm under acromion, and separated deltoideus muscle vertically to touch the fracture,reduced the end of fracture directly and indirectly. PHILOS plate was inserted downward into anterolateral surface of humerus through deltoideus muscle, the distal end and proximal end was fixed by locking screws. The Neer score for shoulder function was evaluated within 1 year after operation.

RESULTSThe operative time ranged from 30 to 70 minutes with an average of 45 minutes. No blood transfusion was required during the operation, and all incisions healed in stage I. All the patients were followed up, and the duration ranged from 6 to 18 months with a mean time of 12.5 months. All the fractures healed up perfectly, and the union time ranged from 6 to 12 weeks. According to Neer criteria for shoulder joint function, 10 patients got an excellent result, 9 good, 2 poor and 1 bad. There were no complications such as axillary nerve injuries, screw loosening, steel plate breakage, dislocation of shoulder joint and necrosis of humeral bone.

CONCLUSIONPHILOS plate through mini-open deltoid-splittin approach for the treatment of proximal humeral fractures has follow advantages: simple recover,minor-injuries and small tissue invasion, which is an ideal method to treat proximal humeral fractures.

Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; instrumentation ; methods ; Shoulder Fractures ; surgery ; Treatment Outcome

OBJECTIVETo establish a simple and efficient method for establishing a mouse model of orthotopic superficial bladder cancer.

METHODSC57BL/6 mice were anesthetized with sodium pentobarbital and catheterized with modified IV catheter (24 G). The mice were intravesically pretreated with HCl and then with NaOH, and after washing the bladders with phosphate-buffered saline (PBS), 100 microl (1 x 10(7)) MB49 cells were infused and allowed to incubate in the bladder for 2 h followed intravesical mitomycin C (MMC) administration. The tumor formation rate, survival, gross hematuria, and bladder weight were determined as the outcome variables, and the pathology of the bladders was observed.

RESULTSInstillation of MB49 tumor cells resulted in a tumor formation rates of 100% in all the pretreated groups while 0% in the control group without pretreatment. MMC significantly reduced the bladder weight as compared to PBS.

CONCLUSIONWe have successfully established a stable, reproducible, and reliable orthotopic bladder cancer model in mice.

Animals ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; Disease Models, Animal ; Female ; Mice ; Mice, Inbred C57BL ; Mitomycin ; pharmacology ; Organ Size ; drug effects ; Urinary Bladder ; drug effects ; pathology ; Urinary Bladder Neoplasms ; pathology

Aim To prepare evodiamine butyryl deriva-tive (EBD) and evodiamine butyryl derivative-loaded solid lipid nanoparticles (EBDLN), and study its re-lease in vitro,and to investigate its in situ gastrointesti-nal absorption. Methods EBD was prepared by a one-step synthetized method, and then EBDLN was prepared by a film dispersion method. Dynamic dialy-sis was used to evaluate drug release in vitro,and sin-gle-pass gastrointestinal perfusion was employed to study the gastrointestinal absorption of EDM,EBD and EBDLN. Results In identical release media, there were identical drug release tendencies of EBD and EB-DLN, but the release rate of EBDLN was faster than EBD. Compared with EDM and EBD, the Kavalues and Pappvalues of EBDLN in every perfusion segment increased significantly. The Kaof EBDLN in stomach, duodenum, jejunum, ileum and colon was 110.14-fold,56.70-fold,51.23-fold,45.70-fold and 127.23-fold of free EDM respectively. The Pappvalue of EB-DLN was 9.74-fold, 4.48-fold, 3.82-fold and 11.3-fold of that of free EDM. Conclusion EBDLN has sustained effect and can enhance the gastrointestinal absorption of EDM and EBD.

Objective:To analyze the distribution and drug sensitivity of pathogens and cerebrospinal fluid (CSF) related measurements in AIDS patients with central nervous system (CNS) infections in Chongqing, so as to provide guidance for etiological diagnosis and rational use of antibiotics in AIDS patients with CNS infections.Methods:A total of 173 AIDS patients with CNS infections were divided into fungal group, Gram-positive bacilli group, Gram-positive cocci group and Gram-negative bacilli group. During the same period, 198 AIDS patients with non-CNS infection visited this hospital were enrolled into the control group. CSF and blood were collected for bacterial culture. The composition and drug resistance of pathogens were analyzed. The levels of CSF related measurements were determined and compared.Results:A total of 173 strains of pathogens were isolated from the CSF of the AIDS patients with CNS infections. The 173 strains included 101 (58.38%) fungi, 39 (22.54%) Gram-positive bacilli, 24 (13.87%) Gram positive cocci and 9 (5.20%) Gram-negative bacilli; 230 strains of pathogens were isolated from the blood of the AIDS patients with non CNS infections. The 198 strains were composed of 107(54.04%) fungi, 65 (32.83%) Gram positive cocci and 26 (13.13%) Gram-negative bacilli. Antifungal sensitivity testing result of Cryptococcus neoformans showed that MIC of amphotericin B, fluorocytosine, fluconazole, voriconazole and itraconazole were≤4, ≤32, ≤8, ≤1 and ≤1 μg/ml. The resistance rate of Mycobacterium tuberculosis to rifampicin was 7.69%. The result of drug sensitivity of coagulase negative staphylococci isolated from patients with CNS infections and non-CNS infections were consistent. There were significant differences among Staphylococcus aureus, Enterobacteriaceae and Acinetobacter. Compared with the control group, the levels of protein were higher and the levels of chloride and glucose were lower in fungal group, Gram-positive bacteria group, Gram-positive coccus group and Gram-negative bacteria group ( t=3.408-9.249, all P<0.011). The levels of protein, adenosine deaminase (ADA) and lactic dehydrogenase (LDH) in Gram-positive bacilli group were significantly higher than those in fungal group, Gram-positive coccus group, Gram-negative bacteria group and control group ( t=3.836-7.686, all P<0.037). Conclusions:The pathogens causing CNS and blood infections in AIDS patients were widely distributed, mainly dominated by fungus. The CSF related measurements varied with different pathogens, so as to assist in the etiological diagnosis of CNS infections.

Objective To investigate the dosimetric effects of different calculation grid size(CGS)in helical tomotherapy(HT)planning system on stereotactic body radiation therapy(SBRT)for non-small cell lung cancer(NSCLC).Methods Nine NSCLC patients receiving radiation therapy for the first time at some hospital from March 2019 to December 2022 were selected as the subjects.SBRT planning was carried out through the HT system with three different CGS plans(Fine,Normal,and Coarse)and the same pitch,modulation factor(MF)and optimization conditions,and the target area indexes of the three CGS plans were compared including conformity index(CI),homogeneity index(HI),dosimetric parameters of the organ at risk(OAR),point dose verification pass rate,treatment time,number of monitor units and Sinograms.SPSS 22.0 was used for statistical analysis.Results For target area HI,there weres significant differences between CGS Fine plan and Coarse plan and between CGS Normal plan and Coarse plan(P＜0.05),while no statistical differences were found between CGS Fine plan and Normal plan(P＞0.05).For target area CI,there were significant differences between CGS Fine plan and Coarse plan(P＜0.05),while no statistical differences were found between CGS Fine plan and Normal plan and between CGS Normal plan and Coarse plan(P＞0.05).For OAR dosimetric parameters,CGS Fine plan and Coarse plan had significant differences in heart Dmax and Dmean,esophageal Dmax and Dmean,V5,V20,V30 and Dmean of the whole lung and affected lung,V5 and Dmax of the affected lung and heart V10 and V30(P＜0.05),CGS Normal plan and Coarse plan had obvious differences in esophageal Dmax(P＜0.05),and the remained dosimetric parameters were not statistically significant(P＞0.05).Fine,Normal and Coarse plans had the point dose verifica-tion pass rates being 0.96％,1.50％and 1.77％,respectively.In terms of treatment time and number of monitor units,there were significant differences between Fine plan and Coarse plan(P＜0.05)while no statistical differences were found between Fine and Normal plans and between Normal and Coarse plans(P＞0.05).Sinograms analyses showed Fine plan had evenly distributed segment color gradient,Coarse plan had areas of very dark and very light color gradients and Normal plan was somewhere in between.Conclusion Low CGS has to be used as much as possible to obtain accurate dose distribution during SBRT planning for NSCLC patients,which contributes to the execution of the radiation therapy plan and the prevention of ad-verse effects.[Chinese Medical Equipment Journal,2024,45(2):52-57]

OBJECTIVE: To investigate the effect of inner-heating acupuncture on apoptosis of chondrocytes and expression of Caspase-3 and Caspase-9 in rats with knee osteoarthritis (KOA). METHODS: A total of 32 rats were divided into a normal group, a model group, a control treatment group and a treatment group by random number grouping method, 8 rats in each one. The rats in the normal group received no intervention. The rats in the remaining three groups adopted modified Videman method to develop KOA model, the ankle joint of left posterior leg was fully extended and fixed with a resin bandage for 6 weeks. After successful modeling, the rats in the model group received no intervention. The rats in the control treatment group were treated with medium-frequency pulse electrotherapy. The rats in the treatment group were treated with inner- heating acupuncture, 30 min each treatment, once a day, five days per week, and totally 3-week treatment was given. After 3 weeks, the damaged cartilage tissue was collected, and HE staining was used to observe the pathological changes of the cartilage tissue of the knee joint. ELISA was used to detect the content of cytochrome-C in the tissue homogenate supernatant. The chondrocytes in damaged cartilage tissue were isolated, flow cytometer was used to detect the changes of apoptosis and mitochondrial membrane potential. The mRNA and protein expression of Caspase-3 and Caspase-9 in chondrocytes were detected by real-time quantitative PCR (qRT-PCR) and Western blot (WB), respectively. RESULTS: Compared with the normal group, the damage of cartilage tissue in the model group was significant, and the expression level of Cyt-C in the homogenate supernatant of damaged cartilage tissue was increased (<0.01); the chondrocyte apoptosis was increased significantly (<0.01); the chondrocyte mitochondrial membrane potential was decreased significantly (<0.01); the mRNA and protein expression of Caspase-3 and Caspase-9 was increased significantly (all <0.01). Compared with the model group, the cartilage injury in the control treatment group and the treatment group was significantly relieved; the expression level of Cyt-C in the supernatant of damaged cartilage tissue homogenate was decreased (both <0.01); the chondrocyte apoptosis was significantly reduced (both <0.01); the chondrocyte mitochondrial membrane potential was increased significantly (both <0.01). Moreover, the mRNA and protein expression of Caspase-3 and Caspase-9 was significantly reduced (all <0.01). Compared with the control treatment group, the treatment group was more effective in the treatment of KOA. CONCLUSION The inner-heating acupuncture could significantly improve the pathological changes of KOA rats, inhibit the apoptosis of chondrocytes, which may be closely related to the suppression of Caspase-3 and Caspase-9 expression.
Animals ; Apoptosis ; Cartilage, Articular ; Caspase 3 ; Caspase 9 ; Chondrocytes ; Heating ; Osteoarthritis, Knee ; Rats

Periarticular fracture of the shoulder is a common type of fractures in the elderly. Postoperative adverse events such as internal fixation failure, humeral head ischemic necrosis and upper limb dysfunction occur frequently, which seriously endangers the exercise and health of the elderly. Compared with the fracture with normal bone mass, the osteoporotic periarticular fracture of the shoulder is complicated with slow healing and poor rehabilitation, so the clinical management becomes more difficult. At present, there is no targeted guideline or consensus for this type of fracture in China. In such context, experts from Youth Osteoporosis Group of Chinese Orthopedic Association, Orthopedic Expert Committee of Geriatrics Branch of Chinese Association of Gerontology and Geriatrics, Osteoporosis Group of Youth Committee of Chinese Association of Orthopedic Surgeons and Osteoporosis Committee of Shanghai Association of Chinese Integrative Medicine developed the Chinese expert consensus on the diagnosis and treatment of osteoporotic periarticular fracture of the shoulder in the elderly ( version 2023). Nine recommendations were put forward from the aspects of diagnosis, treatment strategies and rehabilitation of osteoporotic periarticular fracture of the shoulder, hoping to promote the standardized, systematic and personalized diagnosis and treatment concept and improve functional outcomes and quality of life in elderly patients with osteoporotic periarticular fracture of the shoulder.