Tumor-associated macrophages (TAMs), one of the dominating constituents of tumor microenvironment, are important contributors to cancer progression and treatment resistance. Therefore, regulation of TAMs polarization from M2 phenotype towards M1 phenotype has emerged as a new strategy for tumor immunotherapy. Herein, we successfully initiated antitumor immunotherapy by inhibiting TAMs M2 polarization via autophagy intervention with polyethylene glycol-conjugated gold nanoparticles (PEG-AuNPs). PEG-AuNPs suppressed TAMs M2 polarization in both in vitro and in vivo models, elicited antitumor immunotherapy and inhibited subcutaneous tumor growth in mice. As demonstrated by the mRFP-GFP-LC3 assay and analyzing the autophagy-related proteins (LC3, beclin1 and P62), PEG-AuNPs induced autophagic flux inhibition in TAMs, which is attributed to the PEG-AuNPs induced lysosome alkalization and membrane permeabilization. Besides, TAMs were prone to polarize towards M2 phenotype following autophagy activation, whereas inhibition of autophagic flux could reduce the M2 polarization of TAMs. Our results revealed a mechanism underlying PEG-AuNPs induced antitumor immunotherapy, where PEG-AuNPs reduce TAMs M2 polarization via induction of lysosome dysfunction and autophagic flux inhibition. This study elucidated the biological effects of nanomaterials on TAMs polarization and provided insight into harnessing the intrinsic immunomodulation capacity of nanomaterials for effective cancer treatment.

Valproate is commonly used as a first-line broad-spectrum anti-epileptic drug. Valproate has a narrow therapeutic window, its metabolism is affected by many factors, and it has great individual differences, which makes individualized drug dosage regime needed for valproate. Population pharmacokinetics: is a population analytical method developed in recent years. This paper reviewed the in viva metabolic process and the population pharmacokinetics of valproate in recent years, and analyzed the factors that may affect the metabolism of valproate, including demographic factors, genetic factors and concomitant medications, hoping to provide reference for clinical individualized drug dosage regime.

Objective To study and compare biomechanical properties of a newly developed magnesium AZ31B alloy intramedullary nail (AZ31B) with that of imported Poly-L-lactic acid intramedullary nail (PLLA) and pure titanium rib plate (TPRP), so as to provide scientific evidences for better internal fixation in clinical operation. Methods Forty fresh adult ribs were used and divided into 4 groups randomly. Three groups were made lateral rib fracture in midaxillary line and fixed by AZ31B, PLLA and TPRP, respectively, while the group with normal ribs was used as control. Biomechanical properties of specimens in each group were measured and tested using experimental stress analysis. Results (1) Three-point bending strength of internal fixation with AZ31B was close to that of control group (P>0.05), but significantly different to that of TPRP group and PTRP group (P<0.05); (2) No significant differences in torsional strength were found between AZ31B group, PTRP group and control group (P>0.05), and the torsional strength of both AZ31B and PTRP was superior to that of PLLA (P<0.05). Conclusions The internal fixation with AZ31B is an ideal mode for treating rib fracture since AZ31B has larger flexural strength than PLLA and TPRP, and its torsional strength was close to PTRP and normal ribs. This study provides some support for future research on biomechanical properties of AZ31B.

OBJECTIVETo observe the effects on the lower limbs function after lumbar or sacral nerve root transferring to reconstruct urination function.

METHODSNine patients with bladder dysfunction and normal lower limb function after spinal cord injury were treated with anastomosis the S2 or S3 nerve root with the normal lumbar or sacral nerve root to reconstruct a new bladder artificial reflex arc. Then the alterations on the sensation and motor function of the lower limb after the surgery were observed.

RESULTSMyodynamia of the legs decreased slightly, and the decreasing about half grade of the myodynamia in the plantar flexion of the ankles were detected in 4 of 9 patients with S1 transferring. And the myodynamia recovered 3 months postoperatively. No obvious decreasing of the myodynamia appeared in the other cases.

CONCLUSIONNo obvious effects on the motor function can be found after the single lumbar or sacral nerve root transferring to reconstruct urination function.

Adult ; Exercise ; Female ; Follow-Up Studies ; Humans ; Lower Extremity ; innervation ; physiopathology ; Lumbosacral Region ; Male ; Middle Aged ; Reflex ; Rhizotomy ; Spinal Cord Injuries ; complications ; physiopathology ; Spinal Nerve Roots ; surgery ; Treatment Outcome ; Urinary Bladder ; innervation ; physiopathology ; Urinary Bladder, Neurogenic ; etiology ; physiopathology ; surgery

BACKGROUNDThere are few effective methods for treating injuries to the lower trunk of brachial plexus, and the curative effect is usually poor. The purpose of this study was to provide anatomic references for transferring the brachialis muscle branch of musculocutaneous nerve (BMBMCN) for selective neurotization of finger flexion in brachial plexus lower trunk injury, and to evaluate its clinical curative effects.

METHODSMicroanatomy and measurement were done on 50 limbs from 25 adult human cadavers to observe the origin, branch, type of the BMBMCN and median nerve, as well as their adjacent structures. Internal topographic features of the fascicular groups of the median nerve at the level of the BMBMCN were observed. In addition, the technique of BMBMCN transfer for selective neurotization of finger flexion of the median nerve was designed and tested in 6 fresh adult human cadavers. Acetylcholinesterase (AchE) staining of the BMBMCN and median nerve was done to observe the features of the nerve fibers. This technique was clinically tried to restore digital flexion in 6 cases of adult brachial plexus lower trunk injury. These cases were followed up for 3, 6, 9 and 12 months postoperatively. Recovery of function, grip strength, nerve electrophysiology and muscle power of the affected limbs were observed and measured.

RESULTSThe brachialis muscle was totally innervated by the musculocutaneous nerve (MCN). Based on the Hunter's line, the level of the origin of the BMBMCN was (13.18 +/- 2.77) cm. AchE histochemical staining indicated that the BMBMCN were totally made up of medullated nerve fibers. At the level of the BMBMCN, the median nerve consistently collected into three fascicular groups as shown by microanatomy in combination with AchE stain. The posterior fascicular group was mainly composed of anterior interosseous nerves and branches to the palmaris longus. The technique was tested in six fresh cadavers successfully, except that stoma split occurred in one case. Five of the six cases recovered digital flexion 12 months after operation, and at the same time grip strength, muscle power, and nerve electrophysiology also recovered markedly.

CONCLUSIONSThe technique of transferring the BMBMCN for selective neurotization of finger flexion is anatomically safe and effective, with satisfactory clinical outcomes.

Acetylcholinesterase ; analysis ; Adult ; Brachial Plexus ; anatomy & histology ; injuries ; Brachial Plexus Neuropathies ; surgery ; Clinical Trials as Topic ; Female ; Humans ; Male ; Middle Aged ; Musculocutaneous Nerve ; transplantation ; Nerve Transfer ; methods ; Retrospective Studies

BACKGROUNDActivation and proliferation of hepatic stellate cells (HSC) is essentially involved in the development and progression of hepatic fibrosis. The most potent growth factor for HSC is platelet-derived growth factor receptor (PDGF) and PDGF receptor beta subunit (PDGFR-beta) is the predominant signal transduction pathway of PDGF which is overexpressed in activated HSC. This study investigated the cleavage activity of hammerhead ribozyme targeting PDGFR-beta mRNA in HSC and the effect on biological characteristics of HSC.

METHODSExpression vector of anti-PDGFR-beta ribozyme was constructed and transfected into rat activated HSC with lipofectamin. The positive cell clones were gained by G418 selection. The expression of PDGFR-beta, alpha-smooth muscle actin, and typeI and type III collagen were detected by using Northern blot, Western blot and immunocytochemical staining, respectively. The cell proliferation was determined with MTT colorimetric assay. The cell apoptosis was analyzed by using flow cytometry, acridine orange fluorescence vital staining and transmission electron microscopy.

RESULTSThe expression of PDGFR-beta at mRNA and protein level was markedly reduced in ribozyme-transfected HSC by 49% - 57% (P < 0.05 - 0.01). The proliferation and alpha-smooth muscle actin expression of ribozyme-transfected HSC were significantly decreased (P < 0.05 - 0.01), and the type I and type III collagen synthesis were also reduced (P < 0.01). In addition, the proliferative response of ribozyme-transfected HSC to PDGF BB was significantly inhibited. Otherwise, the apoptotic cells were significantly increased in ribozyme-transfected HSC (P < 0.01), and typical apoptotic cells could be found under transmission electron microscopy.

CONCLUSIONSThe anti-PDGFR-beta ribozyme effectively cleaved the target RNA and significantly inhibited its expression, which blocked the signal transduction of PDGF at receptor level, inhibited HSC proliferation and collagen synthesis, and induced HSC apoptosis. These results suggest that inhibiting PDGFR-beta expression of HSC may be a new target for the therapy of liver fibrogenesis, and ribozyme may be a useful tool for inhibiting PDGFR-beta expression.

Actins ; biosynthesis ; Animals ; Apoptosis ; Cell Proliferation ; Cells, Cultured ; Collagen ; biosynthesis ; Liver ; cytology ; Liver Cirrhosis ; drug therapy ; pathology ; RNA, Catalytic ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Receptor, Platelet-Derived Growth Factor beta ; genetics

Electrocardiography ; Humans ; Lung Abscess ; complications ; Male ; Middle Aged ; Pulmonary Veins ; diagnostic imaging ; Radiography ; Tachycardia, Paroxysmal ; etiology ; surgery

OBJECTIVETo explore the significance and the role of the p53 gene mutation in the exon 4 to 8 in keloid fibroblasts.

METHODSTissue samples from twelve patients with keloid and twelve hyperplastic scar respectively were harvested for in vitro culture of fibroblasts, and normal skin samples from the same patients were employed as the control. Polymerase chain reaction-based single-strained conformational polymorphism (PCR-SSCP) and DNA sequencing were employed to detect p53 gene mutations of the fibroblasts.

RESULTSThe points and frameshift mutations in the exon 4, 5, 6, 7 of p53 gene were identified in 9 of the 12 keloid tissue samples. No p53 gene mutation was detected in all hyperplastic scar and normal skin samples.

CONCLUSIONp53 gene mutation might play an important role in the formation and development of keloids.

Female ; Fibroblasts ; metabolism ; Genes, p53 ; Humans ; Keloid ; genetics ; Male ; Mutation

OBJECTIVETo study the cleavage activity of hammerhead ribozyme targeting at platelet-derived growth factor receptor beta subunit (PDGFR- beta) mRNA in hepatic stellate cells (HSCs) and its effect on the biological characters of HSCs.

METHODSExpression vector of anti-PDGFR- beta ribozyme was constructed and transfected into rat-derived HSC-T6 cells with lipofectin. The positive cell clones were gained by G418 selection. The expression of PDGFR- beta, alpha-smooth muscle actin (alpha-SMA), and type I and type III collagen was detected by means of northern blot, Western blot and immunocytochemical staining respectively. The cell proliferation was determined with MTT colorimetric assay. The cell apoptosis was demonstrated with flow cytometry, acridine orange fluorescence vital staining and transmission electron microscopy.

RESULTSThe expression of PDGFR- beta at mRNA and protein level was markedly reduced in ribozyme-transfected HSCs only 43% to 51% of that in control cells (t > or = 3.957, P < 0.05), and alpha-SMA expression level, type I and type III collagen synthesis ability were also reduced (t > or = 6.790, P < 0.01). The proliferation of ribozyme-transfected HSCs was significantly decreased (t > or = 3.858, P < 0.05), and the proliferation response to PDGF BB was markedly inhibited. However the apoptotic rate was significantly increased in ribozyme-transfected HSCs (chi2 > or = 14.157, P < 0.01), and typical apoptotic cells could be found under transmission electron microscopy.

CONCLUSIONSThe anti-PDGFR- beta ribozyme can be expressed stably in HSCs, cleave the target RNA effectively, inhibit HSCs proliferation and collagen synthesis, and induce HSC apoptosis. The results suggest that inhibiting PDGFR- beta expression in HSCs may be a new therapy for liver fibrosis.

Apoptosis ; drug effects ; Cell Division ; Cells, Cultured ; Hepatocytes ; drug effects ; physiology ; Humans ; Liver ; pathology ; RNA, Catalytic ; pharmacology ; RNA, Messenger ; biosynthesis ; Receptor, Platelet-Derived Growth Factor beta ; antagonists & inhibitors ; metabolism

OBJECTIVETo investigate the expression of heparanase mRNA and its relation with the clinicopathological features and angiogenesis in primary hepatocellular carcinoma (HCC).

METHODSExpression of heparanase mRNA was detected by RT-PCR in 51 HCC lesions, and microvessel density (MVD) was detected by immunohistochemical stain with a factor VIII-related monoclonal antibody.

RESULTSExpression of heparanase mRNA was shown in 49.0% (25/51) HCC lesions. The positive rate of heparanase expression in tumors larger than 3 cm (63.6%, 21/33) was significantly higher than those in smaller tumors (22.2%, 4/18; P < 0.01). Heparanase expression was more frequent in highly invasive tumors (70.0%, 14/20) compared with moderately invasive tumors (46.7%, 7/15) and low invasive ones (25.0%, 4/16; P < 0.05). Moreover, heparanase expression in tumors with high MVD (62.5%, 20/32) was significantly higher than those in tumors with low MVD (26.3%, 5/19; P < 0.05).

CONCLUSIONHeparanase mRNA expression may be important for the growth, invasion and angiogenesis of hepatocellular carcinoma.

Adult ; Carcinoma, Hepatocellular ; blood supply ; enzymology ; pathology ; Female ; Glucuronidase ; biosynthesis ; genetics ; Humans ; Liver ; enzymology ; Liver Neoplasms ; blood supply ; enzymology ; pathology ; Male ; Microcirculation ; pathology ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; enzymology ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Burden