Near-infrared phosphorescent cyclometalated platinum (II) and iridium (III) complexes with metformin moiety: Design and study towards anticancer theranostic agents.

By rational altering the structure of CN auxiliary ligand, Near-infrared (NIR) phosphorescent cyclometalated platinum (II) and iridium (III) complexes with metformin (Met) have been successfully obtained and characterized. The dissociation of Met in aqueous solution can be accelerated by addition of Glutathione (GSH) and alleviated by drop of histidine, accompanied with a significant decay change of deep red phosphorescence. Besides, Pt3 and Ir1 with moiety of btpq mainly selectively targeted and located in Mitochondrial, while Pt1 of ppy and Pt2 with thpy mainly accumulated in endoplasmic reticulum. Moreover, Pt1-3 and Ir1 with metformin moiety all exert a significant enhanced anticancer activity, among them, Pt3 displays ca.66-fold, ca.147-fold and ca.588-fold higher cytotoxicity than cisplatin, Met-free analogue Pt3a and Met. Their relative anticancer mechanism was further investigated, both Pt2 and Pt3 could form covalent interaction with bovine serum albumin (BSA) and effectively induce reactive oxygen species (ROS) generation, arrest of cell cycle, loss of Mitochondrial membrane potential (MMP), display effective anti-metastasis activity and eventually induce apoptosis of cancer cell.

Tandem CAR-T cells targeting FOLR1 and MSLN enhance the antitumor effects in ovarian cancer.

Given the heterogeneity of solid tumors, single-target CAR-T cell therapy often leads to recurrence, especially in ovarian cancer (OV). Here, we constructed a Tandem-CAR targeting two antigens with secretory activity (IL-12) to improve the effects of CAR-T cell therapy. Twenty coexpressed upregulated genes were identified from the GEO database, and we found FOLR1 (folate receptor 1) and MSLN (mesothelin) were specifically and highly expressed in cancer tissues and only 11.25% of samples were negative for both antigens. We observed an increased proliferation rate for these three CAR-T cells, and Tandem CAR-T cells could efficiently lyse antigen-positive OV cells in vitro and secrete higher levels of cytokines than single-target CAR-T cells. More importantly, in vivo experiments indicated that Tandem CAR-T cells markedly decreased tumor volume, exhibited enhanced antitumor activity, and prolonged mouse survival. Furthermore, the infiltration and persistence of T cells in the Tandem-CAR group were higher than those in the MSLN-CAR and Control-T groups but comparable to those in the FOLR1-CAR group. Collectively, this study demonstrated that Tandem CAR-T cells secreting IL-12 could enhance immunotherapeutic effects by reducing tumor antigen escape and increasing T cell functionality, which could be a promising therapeutic strategy for OV and other solid tumors.

Cardiovascular magnetic resonance image analysis and mechanism study for the changes after treatments for primary microvascular angina pectoris.

ABSTRACT: Most cases of primary microvascular angina pectoris (PMVA) are diagnosed clinically, but the etiology and pathological mechanisms are unknown. The effect of routine clinical medications is minimal, and PMVA can progress to serious cardiovascular events. To improve the diagnosis and effective treatment of this disease, this study was designed to diagnose PMVA via cardiovascular magnetic resonance (CMR) and the coronary angiography thrombolysis in myocardial infarction (TIMI) blood flow grade, as well as to analyze vascular endothelial function to elucidate the pathogenesis of PMVA and compare the effects of routine clinical medications.The present randomized controlled trial including a parallel control group will be conducted on 63 PMVA patients in our cardiovascular department. The patients will be selected and randomly divided into the control, diltiazem, and nicorandil groups. The control group will be administered routine drug treatments (aspirin, atorvastatin, betaloc ZOK, perindopril, and isosorbidemononitrate sustained-release tablets). The diltiazem group will be additionally treated with 90 mg qd diltiazem sustained-release capsules. The nicorandil group was additionally given 5 mg tid nicorandil tablets. Coronary angiography will be performed before treatment, the severity and frequency of chest pain will be evaluated before and after 9 months of treatment, and homocysteine and von Willebrand factor levels will be measured. Electrocardiography, echocardiography, dynamic electrocardiography, a treadmill exercise test, and CMR will be performed. Sex, age, body mass index, complications, smoking, and family history will also be recorded. The SPSS19.0 statistical software package will be used to analyze the data. The measurements will be expressed as the mean ±â€Šstandard deviation. Measurement data will be compared between the groups using Student's t-test. A relative number description will be used for the counting data, and the chi-squaretest will be used to compare the groups. A multivariate logistic regression analysis will be performed A P-value < .05 will be considered significant.The direct indices (CMR and coronary angiographic TIMI blood flow grade) may improve after adding diltiazem or nicorandil during routine drug treatments (such as aspirin, statins, and nitrates) in PMVA patients, and indirect indices (homocysteine and von Willebrand factor levels) may be reduced. TRIAL REGISTRATION: Chinese Clinical Trial Registry (http://www.chictr.org.cn/showprojen.aspx?proj=41894), No. CHiCTR1900025319, Registered on August 23, 2019; pre initiation.

Identification and characterization of potential antioxidant components in Isodon amethystoides (Benth.) Hara tea leaves by UPLC-LTQ-Orbitrap-MS.

Isodon amethystoides (Benth.) Hara (IA) tea is a commonly used dietetic Chinese herb and employed for the treatments of tumor and lung abscess. To assess chemical composition and antioxidant capacity of IA leaves extract, a UPLC-LTQ-Orbitrap-MS method and antioxidant tests were used, respectively. 17 compounds were identified including Vinyl caffeate (1), 3,4-dimethoxyphenyl-ß-D-glucopyranoside (2), Rutin (3), Quercetin (4), Loliolide (5), Caffeic acid (6), Rubesanolide D (7), Isorhamnetin (8), Lambertic acid (9), 6, 7-Dehydroroyleanone (10), Dihydrorabdokunmin C (11), Nervosin (12), Quercitrin (13), Vitexin (14), ß-sitosterol (15), Wangzaozin A (16), Amethystonoic acid (17). Among these, 1-14 compounds were novel and have not been reported ever before in IA while component 10 was a novel finding within this genus. Flavonoid components showed better free radical scavenging ability and profound correlation was observed between diterpenoid compounds content and flavonoids activity. Our results provide experimental basis for extraction and separation of chemical constituents of IA which are antioxidant in nature.

Clinical application of percutaneous nephrolithotomy in a patient with kyphoscoliosis.

This case report describes the performance of ultrasound-guided percutaneous nephrolithotomy in a 50-year-old woman who had scoliosis with kyphosis and a history of tuberculosis of the lumbar spine. The operation was performed with the patient under general anesthesia and in the prone position. Residual stones were found in the right lower kidney calyx postoperatively, resulting in a second-phase surgery using the same approach 2 weeks later. All stones were successfully removed during the second surgery. No complications occurred in either operation, and the patient recovered well. This study suggests that ultrasound-guided percutaneous nephrolithotomy is a safe and effective approach in treating renal calculi in patients with scoliosis.

Exosomes derived from mesenchymal stem cells repair a Parkinson's disease model by inducing autophagy.

Parkinson's disease (PD) is a progressively debilitating neurodegenerative condition that leads to motor and cognitive dysfunction. At present, clinical treatment can only improve symptoms, but cannot effectively protect dopaminergic neurons. Several reports have demonstrated that human umbilical cord mesenchymal stem cells (hucMSCs) afford neuroprotection, while their application is limited because of their uncontrollable differentiation and other reasons. Stem cells communicate with cells through secreted exosomes (Exos), the present study aimed to explore whether Exos secreted by hucMSCs could function instead of hucMSCs. hucMSCs were successfully isolated and characterized, and shown to contribute to 6-hydroxydopamine (6-OHDA)-stimulated SH-SY5Y cell proliferation; hucMSC-derived Exos were also involved in this process. The Exos were purified and identified, and then labeled with PKH 26, it was found that the Exos could be efficiently taken up by SH-SY5Y cells after 12 h of incubation. Pretreatment with Exos promoted 6-OHDA-stimulated SH-SY5Y cells to proliferate and inhibited apoptosis by inducing autophagy. Furthermore, Exos reached the substantia nigra through the blood-brain barrier (BBB) in vivo, relieved apomorphine-induced asymmetric rotation, reduced substantia nigra dopaminergic neuron loss and apoptosis, and upregulated the level of dopamine in the striatum. These results demonstrate that hucMSCs-Exos have a treatment capability for PD and can traverse the BBB, indicating their potential for the effective treatment of PD.

miR­15a­3p affects the proliferation, migration and apoptosis of lens epithelial cells.

The present study investigated the effect of microRNA (miR)­15a­3p on the proliferation, migration and apoptosis of lens epithelial cells and its potential mechanism, in order to further elucidate the pathogenesis of age­related cataracts (ARCs). The HLE­B3 human lens epithelial cell line was transfected with miR­15a­3p mimic. Expression of the miR­15a­3p mimic was measured by fluorescence­based reverse transcription­quantitative polymerase chain reaction analysis. Cell proliferation, apoptosis, invasion and migration were investigated using MTT and plate clone formation assays, terminal deoxynucleotidyl transferase dUTP nick end labeling and flow cytometry, and a wound healing assay and Transwell assay, respectively. The protein expression levels of B­cell lymphoma 2 (BCL2) and myeloid cell leukemia sequence 1 (MCL1) were also compared between transfected and wild­type HLE­B3 cells by western blot analysis. The results showed that transfection with the miR­15a­3p mimic significantly suppressed the proliferation of HLE­B3 cells, induced cell apoptosis and increased the proportion of early apoptotic cells. The migration of HLE­B3 cells was significantly inhibited following transfection with miR­15a­3p mimic (P<0.01), whereas cell invasion was unaffected (P>0.05). In addition, reduced protein levels of BCL2 and MCL1 were observed in the miR­15a­3p mimic­transfected HLE­B3 cells (P<0.01). In conclusion, miR­15a­3p may suppress cell proliferation and migration, and induce cell apoptosis in lens epithelial cells through inhibiting the expression of BCL2 and MCL1, which contributes to the onset of ARCs.

Elevated absolute NK cell counts in peripheral blood predict good prognosis in chronic lymphocytic leukemia.

PURPOSE: The aim of this study was to investigate the prognostic significance of the absolute natural killer (NK) cell counts in peripheral blood in patients with chronic lymphocytic leukemia (CLL). METHODS: A total of 273 previously untreated patients with CLL from April 2004 and October 2015 were enrolled into this retrospective study. We analysed the T cell subsets of all patients and figured out the number of NK cells. Comparisons of NK cell count as continuous parameter in different groups were described using Mann-Whitney U test and the Kruskal-Wallis test. Kaplan-Meier method was used to survival analysis, and the Cox proportional hazards models were used for the estimation of prognostic factors. RESULTS: NK cell counts were calculated in 273 therapy-naive CLL patients, and higher number of NK cell was observed in those with Binet stage A/B, ZAP-70 < 20%, normal serum albumin and ß2-microglobulin levels. Using a NK cell count cut-off of 0.40 × 109/L, patients with lower NK cell count (< 0.40 × 109/L) had a significantly shorter overall survival (OS) than those with higher NK cell count (≥ 0.40 × 109/L) (P = 0.0014). Multivariate analysis showed that NK cell counts remained its prognostic value. However, the effect of NK cell count on time to treatment was not significant. CONCLUSIONS: Our results suggest that NK cell count is an independent prognostic marker for OS in patients with CLL and NK cell counts ≥ 0.40 × 109/L can routinely be used to identify patients with favorable survival.

Application of ureterorenoscope and flexible ureterorenoscope lithotripsy in removing calculus from extracorporeal living donor renal graft: a single-center experience.

Here, we reported our clinical application of ureterorenoscope (URS) and flexible URS lithotripsy in stone removal on 10 cases of excised living donor kidney graft. After the extraction of donor kidney by retroperitoneal laparoscopy, the donor graft was perfused with 4 °C HCA solution. Calculus between 2-4 mm were removed intact with lithotomy forceps under direct vision of URS. Larger calculi of >4 mm were fractured with flexible URS combining holmium laser lithotripsy. Fragments of the calculus were extracted with basket extractor and lithotomy forceps. All operations were successful. The operation time was 14-31 min (average 21.2 ± 6.3 min). The kidneys were then transplanted to the recipients using routine procedure. The transplanted kidneys functioned well after transplantation. Gross hematuria resolved 1-4 d after operation (average 2.6 ± 0.9 d). The transplanted kidneys functioned well without early complications such as functional recovery delay and acute graft rejection. The donors and recipients were followed for 12 months. The size of the transplanted kidneys was normal and new stones or urinary obstruction was not seen upon urinary color Doppler ultrasound examination. In conclusion, we believe it is feasible, safe and effective to use URS or flexible URS combining holmium laser lithotripsy on extracorporeal living donor kidney.

Platelet desialylation is a novel mechanism and a therapeutic target in thrombocytopenia during sepsis: an open-label, multicenter, randomized controlled trial.

BACKGROUND: Studies in murine models suggested that platelet desialylation was an important mechanism of thrombocytopenia during sepsis. METHODS: First, we performed a prospective, multicenter, observational study that enrolled septic patients with or without thrombocytopenia to determine the association between platelet desialylation and thrombocytopenia in patients with sepsis, severe sepsis, and septic shock. Gender- and age-matched healthy adults were selected as normal controls in analysis of the platelet desialylation levels (study I). Next, we conducted an open-label randomized controlled trial (RCT) in which the patients who had severe sepsis with thrombocytopenia (platelet counts ≤50 × 109/L) were randomly assigned to receive antimicrobial therapy alone (control group) or antimicrobial therapy plus oseltamivir (oseltamivir group) in a 1:1 ratio (study II). The primary outcomes were platelet desialylation level at study entry, overall platelet response rate within 14 days post-randomization, and all-cause mortality within 28 days post-randomization. Secondary outcomes included platelet recovery time, the occurrence of bleeding events, and the amount of platelets transfused within 14 days post-randomization. RESULTS: The platelet desialylation levels increased significantly in the 127 septic patients with thrombocytopenia compared to the 134 patients without thrombocytopenia. A platelet response was achieved in 45 of the 54 patients in the oseltamivir group (83.3%) compared with 34 of the 52 patients in the control group (65.4%; P = 0.045). The median platelet recovery time was 5 days (interquartile range 4-6) in the oseltamivir group compared with 7 days (interquartile range 5-10) in the control group (P = 0.003). The amount of platelets transfused decreased significantly in the oseltamivir group compared to the control group (P = 0.044). There was no difference in the overall 28-day mortality regardless of whether oseltamivir was used. The Sequential Organ Failure Assessment score and platelet recovery time were independent indicators of oseltamivir therapy. The main reason for all of the mortalities was multiple-organ failure. CONCLUSIONS: Thrombocytopenia was associated with increased platelet desialylation in septic patients. The addition of oseltamivir could significantly increase the platelet response rate, shorten platelet recovery time, and reduce platelet transfusion. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR-IPR-16008542 .

Bioactivities and Chemical Constituents of Essential Oil Extracted from Artemisia anethoides Against Two Stored Product Insects.

The chemical constituents of the essential oil extracted from Artemisia anethoides and the bioactivities of essential oil against Tribolium castaneum and Lasioderma serricorne were investigated. The main components of the essential oil were 1,8-cineole (36.54%), 2-isopropyl-5-methyl-3-cyclohexen-1-one (10.40%), terpinen-4-ol (8.58%), 2-isopropyltoluene (6.20) and pinocarveol (5.08%). The essential oil of A. anethoides possessed contact and fumigant toxicities against T. castaneum adults (LD50 = 28.80 µg/adult and LC50 = 13.05 mg/L air, respectively) and against L. serricorne (LD50 = 24.03 µg/adult and LD50 = 8.04 mg/L air, respectively). The crude oil showed repellent activity against T. castaneum and L. serricorne. Especially, the percentage repellency of essential oil was same level with DEET (positive control) against T. castaneum. The results indicated that the essential oil of A. anethoides had the potential to be developed as insecticide and repellent for control of T. castaneum and L. serricorne.

Cloning and characterization of GST fusion tag stabilized large subunit of Escherichia coli acetohydroxyacid synthase I.

There are three acetohydroxyacid synthase (AHAS, EC 4.1.3.18) isozymes (I, II, and III) in the enterobacteria Escherichia coli among which AHAS I is the most active. Its large subunit (LSU) possesses full catalytic machinery, but is unstable in the absence of the small subunit (SSU). To get applicable LSU of AHAS I, we prepared and characterized in this study the polypeptide as a His-tagged (His-LSU) and a glutathione S-transferase (GST)-tagged (GST-LSU) fusion protein, respectively. The results showed that the His-LSU is unstable, whereas the GST-LSU displays excellent stability. This phenomenon suggests that the GST polypeptide fusion tag could stabilize the target protein when compared with histidine tag. It is the first time that the stabilizing effect of the GST tag was observed. Further characterization of the GST-LSU protein indicated that it possesses the basic functions of AHAS I with a specific activity of 20.8 µmol min(-1) mg(-1) and a Km value for pyruvate of 0.95 mM. These observations imply that introduction of the GST fusion tag to LSU of AHAS I does not affect the function of the protein. The possible reasons that the GST fusion tag could make the LSU stable are initially discussed.

Comparison of gene expression of the oncogenic Wnt/ß-catenin signaling pathway components in the mouse and human epididymis.

ß-catenin is an integral part of the Wnt signaling pathway and has been linked to tumorigenesis and multiple developmental processes. The high ß-catenin expression with low tumor incidence in the human epididymis is thus intriguing. In the present study, the ß-catenin gene and protein was found to be highly expressed in the murine caput epididymidis, and the protein mainly localized along the lateral plasma membranes of adjacent epithelial cells throughout both human and mouse epididymides. Furthermore, the adult mouse epididymis was found to express almost all the Wnt/ß-catenin signaling pathway genes that were determined previously by our group in the human organ. Despite the differences in epididymal structure, the similar location of ß-catenin and the high concordance of this pathway's components' gene expression in both the adult human and mouse epididymides make the mouse a suitable animal model for studying the anti-tumor mechanism of the epididymis. In addition, both the mRNA and protein expression of ß-catenin shared a similar spatial expression as the mRNA of Ros1, a proto-oncogene and a key developmental regulator of the initial segment of the mouse epididymis. The observations on the parallel temporal expression of ß-catenin and Ros1 during postnatal development raise the possibility that the canonical Wnt signaling pathway has an additional role in the postnatal development of mouse epididymis.

Stable radio-frequency phase distribution over optical fiber by phase-drift auto-cancellation.

We report a new radio-frequency (RF) phase stabilization approach for long-haul optical fiber distribution. The phase drift of an RF signal induced by fiber-length variations can be canceled out automatically via RF mixing without using active phase discrimination and dynamic phase tracking. A key significance of our approach is that no assistant local oscillator (LO) signal is needed. Consequently, frequency estimation of the received RF signal, as well as frequency locking between the LO and the received RF signal, is no longer required, which simplifies the system. A proof-of-concept experiment shows that the phase drift of the received RF signal at 9.6 GHz is significantly reduced using the proposed method. The root mean square (RMS) timing jitter is 0.76 ps when a tunable optical delay line (TODL) inserted between the remote antenna unit (RAU) and local station is changed from 0 to 300 ps.

All-optical frequency upconversion for radio-over-fiber applications based on cross-gain modulation and cross-polarization modulation in a semiconductor optical amplifier.

We propose a novel all-optical frequency upconversion technique for radio-over-fiber (RoF) systems based on cross-gain modulation (XGM) and cross-polarization modulation (XPolM) in a semiconductor optical amplifier (SOA). A local oscillator signal is carried onto a continuous wave probe beam with orthogonally polarized single-sideband (SSB) modulation using a polarization modulator and a tunable optical filter. The intermediate frequency signal carried by a pump beam is only intensity modulated onto the sideband of the probe beam while the carrier of the probe beam is unmodulated thanks to the joint use of the XGM and XPolM effects in the SOA. This SSB upconversion scheme is inherently free from the chromatic-dispersion-induced power fading after transmission over single-mode fiber. The proposed scheme is theoretically analyzed and experimentally verified.

Zhuanggu Jianxi Decoction () limits interleukin-1 ß-induced degeneration chondrocytes via the caveolin-p38 MAPK signal pathway.

OBJECTIVE: To evaluate the effect of Zhuanggu Jianxi Decoction (, ZGJXD) on interleukin-1 ß (IL-1 ß)-induced degeneration of chondrocytes (CDs) as well as the activation of caveolin-p38 mitogen-activated protein kinase (MAPK) signal pathway, investigating the possible molecular mechanism that ZGJXD treats osteoarthritis. METHODS: Serum pharmacology was applied in the present study, where ZGJXD was orally administrated to New Zealand rabbits and then ZGJXD containing serum (ZGJXD-S) was collected for following in vitro experiments. CDs were isolated aseptically from New Zealand rabbits and then cultured in vitro. Upon IL-1 ß stimulation, the degeneration of CDs was verified by inverted microscope, toluidine blue stain and type II collagen immunocytochemistry. After IL-1 ß-stimulated CDs were intervened with blank control serum, ZGJXD-S, together with or without SB203580 (a specific inhibitor of p38 MAPK) for 48 h, caveolin-1 protein expression and the phosphorylation level of p38 were determined by Western blotting, and the mRNA expression of IL-1 ß, tumor necrosis factor α (TNF-α), matrix metalloproteinase 3 (MMP-3) and MMP-13 were examined by real-time polymerase chain reaction. RESULTS: IL-1 ß stimulation induced degeneration of CDs, increased caveolin-1 expression and p38 phosphorylation, up-regulated the mRNA level of IL-1 ß, TNF-α, MMP-3 and MMP-13. However, the IL-1 ß-induced activation of caveolin-p38 signaling and alteration in the expression of p38 downstream target genes were suppressed by ZGJXD-S and/or SB203580 in CDs. CONCLUSION: ZGJXD can prevent CDs degeneration via inhibition of caveolin-p38 MAPK signal pathway, which might be one of the mechanisms that ZGJXD treats osteoarthritis.

[Effect of naringin of Drynaria Rhizome, a Chinese medical component of Zhuanggu Jianxi Recipe containing serum on caveolin-p38MAPK signal pathway in IL-1ß induced rabbit degenerated chondrocytes].

OBJECTIVE: To observe the effect of naringin of Drynaria Rhizome, a Chinese medical component of Zhuanggu Jianxi Recipe (ZJR) containing serum on caveolin-p38MAPK signal factors (such as caveolin-1, p-p38, p-ATF-2, IL-1ß, and TNF-α) in IL-1ß induced rabbit degenerated chondrocytes, and further to explore its mechanism for protecting articular cartilages. METHODS: Naringin of Drynaria Rhizome was obtained and analyzed by HPLC-TOF/MS. Four weeks old New Zealand rabbits were killed and their bilateral knee joints were isolated aseptically. CDs were isolated and then cultured in vitro. The second generation of CDs were used for later experiment. The effect of naringin on CDs proliferation was detected by MTT assay. The effect of naringin on the expression of IL-1ß-induced collagen II in CDs was detected by immunohistochemical method. The effect of naringin on caveolin-1, p-p38, and p-ATF-2 protein in IL-1ß-induced CDs was detected by Western blot. The effect of naringin on mRNA expression of IL-1ß and TNF-α in IL-1ß-induced CDs was detected by RT-PCR. RESULTS: The appearance time of naringin in flow graphs of naringin standard solution and ZJR containing serum was 23.5 min, and the molecular weight ranged between 581.0 and 581.5 m/z. Naringin could promote the proliferation of CDs, and inhibit the effect of IL-1ß on collagen II in CDs. Compared with the model group, naringin could reduce the expression of caveolin-1, p-p38, p-ATF-2, IL-1ß, and TNF-α in IL-1ß induced CDs (P < 0.05), which was approximate to the level of the normal group. CONCLUSIONS: Naringin could not only promote the proliferation of CDs, but also protect IL-1ß-induced CDs. Its mechanism might be associated with decreasing the expression of caveolin-1, p-p38, and p-ATF-2 proteins, inhibiting caveolin-p38MAPK signal pathway, and further reducing mRNA expression of IL-1ß and TNF-α in the downstream of caveolin-p38MAPK signal pathway.

Topical administration of peroxiredoxin-6 on the cornea suppresses inflammation and neovascularization induced by ultraviolet radiation.

PURPOSE: To investigate the effect of topical administration of peroxiredoxin-6 (PRDX6) on ultraviolet-induced corneal injury. METHODS: Corneal transparency and neovascularization were observed with a slit-lamp microscope and hematoxylin and eosin staining. The oxidative damage was determined with a commercial malondialdehyde (MDA) kit. The expressions of PRDX6, polymorphonuclear neutrophil (PMN), vascular endothelial growth factor (VEGF), and pigment epithelium-derived factor (PEDF) were determined by immunohistochemistry and Western blot. The expressions of genes related with antioxidant defense systems and cell apoptosis were detected by RT-PCR. RESULTS: The irradiated corneas appeared opaque and had high levels of MDA. Peripheral neovascularization and neutrophils appeared in the control and buffer-treated groups (with no treatment or PRDX6 diluent, respectively), whereas they were significantly suppressed in the PRDX6-treated group. The MDA content of the corneas in the PRDX6-treated group was significantly lower than that of the control and buffer-treated groups (P < 0.05). In the PRDX6-treated group the immunoreactivity of VEGF was lower, and that of PEDF was higher, than that in the control and buffer-treated groups. In addition, there were expression correlations between PRDX6 and PMN, VEGF, PEDF. The expressions of genes related with antioxidant defense systems and cell apoptosis were significant different between buffer- and PRDX6-treated groups (P < 0.05). CONCLUSIONS: The topically administered PRDX6 maintained the homeostasis of corneal cells, reduced inflammation, and suppressed neovascularization and apoptosis under ultraviolet irradiation.

[Experimental study on the recycling of denatured acellular dermal matrix after burn].

OBJECTIVE: To explore the feasibility of burn denatured acellular dermal matrix (DADM) as dermal substitute in repairing wounds. METHODS: (1) Nine Wistar rats received a deep partial-thickness scald on the back. Full-thickness wounded skin was collected on post scald day (PBD) 1, 2, and 3 (with 3 rats at each time point), and it was treated with 2.5 g/L trypsin/0.5% Triton X-100 to remove cells to prepare DADM, respectively called DADM-1 d, DADM-2 d, and DADM-3 d. Another 3 rats without scald injury were treated with the same method as above to prepare acellular dermal matrix (ADM) to serve as control. Gross and histological observations and microbiological and biomechanical tests, including ultimate tensile strength, maximum tension, stretched length at breaking, stress-strain relationship, were conducted for the resulting ADM and DADM. (2) Another 64 rats were divided into ADM group and DADM-1 d, DADM-2 d, and DADM-3 d groups according to the random number table, with 16 rats in each group. A skin flap in size of 2.0 cm×1.8 cm was raised on the back of each rat. The above-mentioned ADM, DADM-1 d, DADM-2 d, and DADM-3 d were cut into pieces in the size of 1.8 cm×1.5 cm, and they were respectively implanted under the skin flaps of rats in corresponding group. At post surgery week (PSW) 1, 3, 5, or 9, 4 rats in each group were used to observe wound healing condition and change in implants with naked eye, and histological observation of the implants was conducted. Data were processed with one-way analysis of variance and t test. RESULTS: (1) The freshly prepared DADM was milky white, soft in texture with flexibility, but poor in elasticity as compared with ADM. No epithelial structure or cellular component was observed in ADM or DADM under light microscope. Collagen fibers of DADM were seen to be thickened unevenly and arranged in disorder and eosinophilic. All microbiological results of DADM were negative. There was no statistically significant difference among DADM-1 d, DADM-2 d, and DADM-3 d in levels of ultimate tensile strength, maximum tension, stretched length at breaking, and stress-strain relationship (with F values from 0.088 to 3.591, P values all above 0.05). Values of the above-mentioned four indexes were the highest in DADM-3 d, they were respectively (13.0 ± 2.4) MPa, (61 ± 4) N, (173 ± 7)%, (45.7 ± 2.0)%. Values of the four indexes of ADM were respectively (19.0 ± 2.6) MPa, (95 ± 4) N, (201 ± 5)%, (62.5 ± 2.2)%, which were higher than those of DADM-1 d, DADM-2 d, and DADM-3 d (with t values from 6.424 to 17.125, P values all below 0.01). (2) No exudate or swelling in the wounds of rats, and no contraction or curling of implants were observed in every group at PSW 1, but inflammatory cells infiltration and Fbs inward migration were observed in the wound. At PSW 3, the growth of hair was normal in the wound in DADM-1 d, DADM-2 d, and ADM groups, but few and scattered hair grew in DADM-3 d group. The inflammatory cells decreased, while Fbs increased, and new capillaries were found to grow inwardly in each group. The decrease in inflammatory cells was slightly delayed in DADM-3 d group. At PSW 5, hair growth became normal, and implants shrank and thinned with fiber membrane wrapped densely and bundles of ingrowing large caliber blood vessels in all groups. The dermal matrix in each group merged with the surrounding normal tissue. At PSW 9, ADM and DADM became white, thin, and soft tissue sheet which was closely connected with the inner side of the flap. There was no infiltration of inflammatory cells in implants in either group. The collagen fibers arranged regularly and densely, and they were integrated with normal collagen tissue. CONCLUSIONS: The burned DADM does not have obvious immunogenicity, but with good biocompatibility. It is prospective to become as a dermal substitute in repairing wounds.

Research progress in transient receptor potential vanilloid 1 of sensory nervous system.

The transient receptor potential vanilloid subfamily member 1 (TRPV1) is a protein mainly expressed in sensory neurons and fibers, such as in trigeminal ganglion and dorsal root ganglion, and has been indicated to be involved in several physiological and pathological processes. Studies on thermal activation have revealed that phosphorylation is involved in TRPV1 activation and 2 putative phosphorylation sites, Ser residues 502 (Ser-502) and Ser residues 800 (Ser-800), have been recently confirmed to possess the capability of resensitizing TRPV1. In addition to acidification, alkalization has also been proved to be a highly effective stimulator for TRPV1. TRPV1 could be regulated by various physical and chemical modulators, as well as the chronic pain. TRPV1 plays a crucial role in the transmission of pain signals, especially under inflammation and the neoplasm conditions, and it can also modulate nociceptive afferents by reinforcing morphine tolerance. The present review mainly focused on the structural and functional complexities of TRPV1, together with its activation and modulation by a wide variety of physical and chemical stimuli. Its pharmacological manipulation (sensitization/desensitization) and therapeutical targets were also discussed.