[Clinical application of Neuroform Atlas stent-assisted coiling in the treatment of unruptured wide-neck intracranial aneurysms].

OBJECTIVE: To assess the safety and efficacy of Neuroform Atlas stent used in treatment of unruptured wide-neck intracranial aneurysms. METHODS: Clinical data of 62 patients with unruptured wide-neck intracranial aneurysms undergoing Neuroform Atlas stent-assisted coiling from August 2020 to September 2021 were retrospectively analyzed. There were 64 aneurysms in those 62 patients. Among them, 25 aneurysms were located at the bifurcation of M1 segment on middle cerebral artery, 16 at the anterior communicating artery, 10 at the C7 segment of internal carotid artery, 5 at the C6 segment of internal carotid artery, 4 at the apex of basilar artery, 3 at the A3 segment of anterior cerebral artery, and 1 at the M2 segment of middle cerebral artery. All the patients underwent Neuroform Atlas stent-assisted coiling, including 49 patients with single stent assisted coiling and 15 patients with dual stents assisted coiling (14"Y"style and 1"X"style). After the procedure, the immediate DSA was performed to evaluate the status of aneurysm occlusion and the parent artery patency. The clinical follow-up was performed 3 months after the operation and evaluated based on the modified Rankin Scale(mRS).DSA image was reviewed at 6 months after operation and Raymond grading scale was used to assess the status of aneurysm occlusion and the parent artery patency. RESULTS: A total of 62 patients with 64 aneurysms were all achieved technical success(100%).The immediate post-procedural Raymond scale was assessed, including Raymond Ⅰ in 57 aneurysms(89.1%, 57/64), Raymond Ⅱ in 6 aneurysms(9.3%, 6/64) and Raymond Ⅲ in 1 aneurysm(1.6%, 1/64). The peri-procedural complications rate was 4.8%(3/62), 2 patients developed intraoperative thrombosis and 1 patient suffered from local subarachnoid hemorrhage. Among them, 55 patients obtained 3 months clinical follow-up after operation and all the patients had good outcomes (mRS≤2), 50 patients with 52 aneurysms were followed up with DSA 6 months after operation, including Raymond Ⅰ in 45 aneurysms(86.5%, 45/52), Raymond Ⅱ in 4 aneurysms(7.7%, 4/52) and Raymond Ⅲ in 3 aneurysms(5.8%, 3/52). CONCLUSION: Neuroform Atlas stent for the treatment of unruptured wide-neck intracranial aneurysms has high safety and good efficacy, and has its advantages over other traditional stents.

Incidence and familial clustering of infantile haemangiomas: a multicentre study.

BACKGROUND: The exact incidence of infantile haemangiomas (IH) in the Chinese population is still unknown. A positive family history of IH was considered as a risk factor for the development of IH. OBJECTIVES: This study aims to investigate the incidence of IH in the Chinese population and the mechanism of family history increases the risk for IH development. METHODS: A total of 2489 women and their newborns were enrolled in the prospective study. All newborns were followed up for 12 months to determine whether they developed IH. In addition, 213 IH probands and their 174 siblings were enrolled in the study. The incidence of IH in siblings of the IH probands was investigated. Information regarding risk factors for IH and demographic data were collected on all children. RESULTS: Of the 2572 newborns, 58 IH were identified in 56 (2.2%) newborns. The majority of IH were located on the trunk (46.6%). Siblings of the IH probands were at increased risk for the development of IH (P = 0.024, relative risk 2.451), and the occurrence of prenatal risk factors for IH(P = 0.003) compared with the general population. CONCLUSIONS: Our study showed that the incidence of IH is 2.2% in the Chinese population. Siblings of the individuals with IH were at increased risk for the development of IH may be related to the family clustering of prenatal risk factors for IH. Further exploration of the mechanisms and common features of these prenatal risk factors may help to disclose the origin and pathogenesis of IH.

Supraclavicular lymph node recurrence after radical surgery: is epidermal growth factor receptor a predictive marker?

The aim of this study was to evaluate the rare postoperative supraclavicular metastasis originating from oral squamous cell carcinoma (OSCC) and to discuss epidermal growth factor receptor (EGFR) as a potential predictive marker. Tumour specimens of OSCC patients divided into three groups were included: supraclavicular metastasis (n = 8), conventional cervical metastasis (n = 28), no metastasis (n = 48). Basic information and EGFR expression were compared among these groups and the data were analysed to identify potentially related risk factors for supraclavicular metastasis. In the supraclavicular metastasis group (n = 8), all primary tumours were T1-T2 and located in the tongue and buccal region; five of eight cases were pathologically N0. The median interval from the primary tumour resection to the development of supraclavicular metastases was 21.5 months. All related deaths (5/8) occurred within 2 years. In the supraclavicular metastasis group, EGFR expression was highest in the supraclavicular metastases, followed by cervical lymph nodes, and was lowest in the primary tumours (P = 0.39). In contrast, in the conventional metastasis group and the N0 group, EGFR expression was higher in the primary tumours than in the lymph nodes (P < 0.01). Supraclavicular metastasis of OSCC is infrequent and associated with a poor prognosis. EGFR might predict the occurrence of supraclavicular metastasis.

HPV16 infection promotes an M2 macrophage phenotype to promote the invasion and metastasis of esophageal squamous cell carcinoma

Objectives High-risk human papillomavirus (HR-HPV) is an important risk factor for esophageal cancer. Macrophages constitute a crucial immune medium for regulating HPV-related tumors; however, the specific regulatory mechanisms remain unknown. Therefore, the purpose of our current study was to investigate the mechanism by which HPV16E6 regulates macrophages to promote the invasion and metastasis of esophageal cancer. Methods HPV16E6 infection was detected by polymerase chain reaction. Immunohistochemistry was used to verify the distribution of tumor-associated macrophages (TAMs) and MMP-9 expression in esophageal squamous cell carcinoma tissues (ESCCs), and cancer adjacent normal tissues (CANs) from Kazakh patients. ESCC cells were transfected with a plasmid over-expressing HPV16E6 and non-contact cocultured with macrophages. Results The infection rate of HPV16E6 in Kazakh ESCCs was clearly higher than that in CANs (P < 0.05). The density of CD163-positive TAMs was significantly positively correlated with HPV16E6 infection in ESCCs (P < 0.05). After coculturing macrophages and EC9706 cells transfected with the HPV16E6 plasmid, the phenotype of macrophages transformed into M2 macrophages. The migration and invasion ability of ESCC cells were higher in the HPV16E6-transfected and coculture group than in the HPV16E6 empty vector-transfected and non-cocultured HPV16E6-transfected groups (all P < 0.05). The density of M2-like TAMs in ESCCs was positively correlated with the level of MMP-9 expression. MMP-9 expression in the HPV16E6-ESCC coculture macrophages group was substantially higher than that in controls (all P < 0.05). Conclusions HPV16 infection mediates tumor-associated macrophages to promote ESCC invasion and migration (AU)

HPV16 infection promotes an M2 macrophage phenotype to promote the invasion and metastasis of esophageal squamous cell carcinoma.

OBJECTIVES: High-risk human papillomavirus (HR-HPV) is an important risk factor for esophageal cancer. Macrophages constitute a crucial immune medium for regulating HPV-related tumors; however, the specific regulatory mechanisms remain unknown. Therefore, the purpose of our current study was to investigate the mechanism by which HPV16E6 regulates macrophages to promote the invasion and metastasis of esophageal cancer. METHODS: HPV16E6 infection was detected by polymerase chain reaction. Immunohistochemistry was used to verify the distribution of tumor-associated macrophages (TAMs) and MMP-9 expression in esophageal squamous cell carcinoma tissues (ESCCs), and cancer adjacent normal tissues (CANs) from Kazakh patients. ESCC cells were transfected with a plasmid over-expressing HPV16E6 and non-contact cocultured with macrophages. RESULTS: The infection rate of HPV16E6 in Kazakh ESCCs was clearly higher than that in CANs (P < 0.05). The density of CD163-positive TAMs was significantly positively correlated with HPV16E6 infection in ESCCs (P < 0.05). After coculturing macrophages and EC9706 cells transfected with the HPV16E6 plasmid, the phenotype of macrophages transformed into M2 macrophages. The migration and invasion ability of ESCC cells were higher in the HPV16E6-transfected and coculture group than in the HPV16E6 empty vector-transfected and non-cocultured HPV16E6-transfected groups (all P < 0.05). The density of M2-like TAMs in ESCCs was positively correlated with the level of MMP-9 expression. MMP-9 expression in the HPV16E6-ESCC coculture macrophages group was substantially higher than that in controls (all P < 0.05). CONCLUSIONS: HPV16 infection mediates tumor-associated macrophages to promote ESCC invasion and migration.

[Effect of pedicled thoracodorsal artery perforator flap on the repair of moderate to severe axillary scar contracture deformity].

Objective: To investigate the effect of pedicled thoracodorsal artery perforator (TDAP) flap on the repair of axillary moderate to severe scar contracture deformity. Methods: From January 2012 to January 2017, 29 patients with axillary moderate to severe scar contracture deformity were admitted to the the Second Xiangya Hospital of Central South University, including 18 females and 11 males, aged 14-42 years. There were 3 patients with cicatricial contracture deformity of bilateral axillas and 26 patients with cicatricial contracture deformity of unilateral axilla. After relevant preoperative examinations were completed and basic diseases were controlled, axillary scar was removed or released under the anesthesia of endotracheal intubation. The areas of wounds ranged from 7.5 cm×5.0 cm to 21.0 cm×8.5 cm after the operation. The pedicled TDAP flaps were used to repair the defects, which were thinned based on demand of the recipient sites before being transferred. The areas of flaps ranged from 9.0 cm×6.0 cm to 22.0 cm×10.0 cm. The donor sites were sutured directly. The status of thinned flaps, the survival of flaps after the operation and during follow-up, and the shoulder joint function during follow-up were observed. Results: Thirty-two pedicled TDAP flaps were harvested for repairing the defects. Among them, 14 flaps were transferred directly without thinning and the thickness of the flaps ranged from 9.0 mm to 15.0 mm, with average thickness of 13.6 mm.While the other 18 flaps were thinned, and the thickness of the thinned flaps ranged from 5.0 mm to 8.0 mm, with average thickness of 7.5 mm. The distal parts of 3 flaps in 3 patients showed small size of blackening or necrosis within 72 hours after the operation, and 2 of them were thinned and the other one was not. Finally, the 3 flaps were healed after hyperbaric oxygen therapy, dressing change, or other treatments. One flap occurred vein congestion 8 hours after the operation caused by pressure on the pedicle, and the color of the flap turned back to normal after the pressure was relieved. The rest of the flaps survived well. All the patients were followed up for 9 to 36 months, with an average of 18 months. All the flaps survived well, the color of the flaps was nearly the same as the recipient sites, and none of the flaps developed obvious contraction. The shoulder joint function of all patients was significantly improved compared with that before operation, with abduction angles of shoulder joints ranged from 90.0-145.0°, with an average of 130.0°. Conclusions: Pedicled TDAP flap is an relatively ideal choice for the repair of moderate to severe axillary scar contracture deformity, and better results will be achieved if the flaps are thinned to a appropriate thickness according to the condition of axillary defects.

[Clinical application of submental artery perforator flap in oral and maxillofacial defect restoration after tumor resection].

Objective: To summarize the clinical anatomical features and surgical technique of the submental artery perforator flap (SMAPF), and to evaluate the outcome and value of the flap for oral cavity reconstruction after cancer ablation. Methods: A total of 56 patients with oral cancer were included in this study. The modified SMAPF excluded the anterior belly of the digastric muscle and submental fatty tissue. The primary sites of malignancy were buccal mucosa (n=24), tongue (n=21), mandibular gingiva (n=6), mouth floor (n=3), soft palate (n=2). Results: The flap size varied from 3 cm×5 cm to 5 cm×12 cm. Four flaps presented mild venous congestion, which was salvaged with conservative measures such as acupuncture and blood letting. Overall flap survival rate was 100%. The SMAPF with septocutaneous perforator was used in 49 cases and with musculocutaneous perforator in seven cases. External jugular vein and internal jugular vein provides venous drainage were applied in half cases respectively. All the patients showed a good recovery of tongue mobility and mouth opening with a follow-up of 3-41 months. Local recurrence was detected in one patient 4 months after operation. Conclusions: The SMAPF is a reliable flap for oral cavity reconstruction with outstanding functional and aesthetic outcomes.

Carbon monoxide releasing molecule-2 suppresses proliferation, migration, invasion, and promotes apoptosis in non-small cell lung cancer Calu-3 cells.

OBJECTIVE: Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, which is the leading cause of cancer-related morbidity and mortality worldwide. The carbon monoxide-releasing molecules (CO-RMs) are transition metal carbonyls with the capacity to release carbon monoxide (CO). The aims of our study were to assess the effects and underlying mechanisms of CO-releasing molecules-2 (CORM-2) on proliferation, migration, invasion and apoptosis in NSCLC cells, and to evaluate its potential application for lung cancer. MATERIALS AND METHODS: NSCLC cells Calu-3 were treated with CORM-2, negative control and blank control. Cell proliferation, migration and invasion were assessed by cell Counting Kit-8 (CCK-8), scratch assay and matrigel invasion chamber experiment, respectively. Apoptosis was measured by flow cytometry. Real-time PCR and Western blot were applied to examine the expression of apoptosis-related molecules on mRNA and protein levels. RESULTS: CORM-2 markedly attenuated proliferation, migration and invasion of Calu-3 cells. CORM-2 treatment also significantly reduced the ratio of B cell lymphoma 2 (Bcl-2)/B cell lymphoma 2 associated X protein (Bax) while increased expression of caspase-3 and cytochrome c. The optimal dose of CORM-2 for Calu-3 cells was 100 µM. CONCLUSIONS: CORM-2 modulates biological functions of NSCLC cells and may provide a novel therapeutic strategy for lung cancer.

Vertical platysma myocutaneous flap reconstruction for oral defects using three different incision designs: experience with 68 cases.

This study evaluated the effects of three different incision designs for the vertical platysma myocutaneous flap (VPMF): apron, MacFee, and T-shaped. This flap was used for the reconstruction of intraoral defects following cancer ablation in selected patients. Sixty-eight cases of VPMF reconstruction were assessed: the apron incision was used in 28, MacFee incision in 22, and T-shaped incision in 18. With regard to postoperative outcomes, there were 26 cases of flap survival and two of partial necrosis with the apron incision; 20 of survival and two of partial necrosis with the MacFee incision; 15 of survival and three of partial necrosis with the T-shaped incision. Success rates were 92.9%, 90.9% and 83.3%, respectively, for VPMF with the apron, MacFee, and T-shaped incisions. A wound healing disturbance in the neck was seen in three cases of VPMF with the apron incision and one case with the MacFee incision. The MacFee incision had the best aesthetic effect, and the postoperative neck scar was more obvious for the T-shaped incision. It is recommended that VPMF with the MacFee or apron incision be used for the reconstruction of larger buccal mucosa and floor of the mouth defects, while VPMF with the T-shaped incision should be used for smaller intraoral defects, especially tongue defects of the lateral surface.

Differences in photoreceptor recovery among patients and between different parts of the posterior pole in Vogt-Koyanagi-Harada disease.

PurposeTo investigate the recovery of photoreceptors following the treatment in Vogt-Koyanagi-Harada (VKH) disease.Patients and methodsThis was a retrospective study. We enrolled 28 patients with VKH (56 eyes). The clinical and optical coherence tomography (OCT) findings were recorded for 12 months after treatment. The patterns of photoreceptor recovery on OCT were defined: pattern F group=Foveal photoreceptor recovery visible first; pattern E group=Extrafoveal photoreceptor recovery visible first; and pattern S group=Simultaneous foveal and extrafoveal photoreceptor recovery.ResultsPhotoreceptor recovery varied in different parts of the fundus among patients. Among the 56 eyes, the ellipsoid zone (EZ) recovery of 10 eyes and the interdigitation zone (IZ) recovery of 17 eyes belonged to pattern F group. In most eyes (46 eyes for EZ and 26 eyes for IZ), the recovery of these structures were pattern S. Only in 10 eyes, the recovery of IZ was pattern E. The different patterns of recovery correlated with how promptly the patients had been treated and with the anatomical and visual outcomes at 12 months. Patients in pattern F group were characterized by delayed treatment, delayed recovery of EZ or IZ, and a less favourable prognosis at 12 months relative to other patients, while those in pattern E group had the most prompt treatment and recovery as well as a more favourable outcome at 12 months.ConclusionsIn VKH patients with delayed treatment, foveal photoreceptors tended to recover more rapidly than photoreceptors in other regions.

Comparing the ability of luminex xMAP(®) salmonella serotyping assay and traditional serotyping method for serotyping salmonella isolated from southern Chinese population.

AIMS: To investigate whether the Luminex xMAP(®) Salmonella Serotyping Assay (xMAP SSA) is applicable to serotype Salmonella from humans in southern China. METHODS AND RESULTS: Two hundred and five Salmonella isolates from diarrhoea patients were serotyped by xMAP SSA in parallel with the traditional serotyping. Forty serotypes were identified among 205 isolates; the most prevalent serotypes identified were Salmonella Enteritidis, Salmonella Stanley, Salmonella I 4,5,12:i:-, and Salmonella Typhimurium. One hundred and ninety-five (95·1%, 195/205) isolates were serotyped completely by xMAP SSA, while 10 stereotypes were partially detected as they were not included in the assay. The xMAP SSA correctly identified 192 (98·5%, 192/195) isolates. Five nonmotile and three monophasic strains, which possessed flagellar antigen genes that weren't expressed, were completely serotyped by xMAP SSA; however, these isolates were left undetected by the traditional method. CONCLUSION: The xMAP SSA used in the study is a microsphere-based, molecular assay that could rapidly and accurately serotype Salmonella. It is suitable to identify the serovars of Salmonella in southern China. SIGNIFICANCE AND IMPACT OF THE STUDY: The xMAP SSA, with high-throughput characteristics, provides an accurate and rapid serotyping system that dramatically strengthens the capability of clinical and public health laboratories for Salmonella serotyping.

The study of prevalence rate, and clinical characteristics of high altitude deterioration.

OBJECTIVE: This study was aimed to investigate the prevalence rate and clinical characteristics of high altitude deterioration (HADT), which would provide a scientific basis for the diagnosis and prevention of HADT. SUBJECTS AND METHODS: A total of 175 subjects, who had migrated to a high altitude (4516 m) for more than 1 year, were investigated. A questionnaire survey based on the symptoms of HADT was conducted, and 117 subjects were determined to have HADT according to the diagnostic criteria of HADT. To explore the clinical characteristics of HADT, 117 HADT patients and 31 healthy individuals were assigned to HADT patient group and healthy control group, respectively. Their body mass indexes (BMIs), blood rheology, full blood count (including hemoglobin concentration, leukocyte count, neutrophil count, lymphocyte count and platelet count), blood pressure, heart rate, oxygen saturation as well as left ventricular ejection fraction (LVEF) and fraction shortening (LVFS) were assessed. RESULTS: The prevalence rate of HADT was 66.9% at a high altitude of 4516 m. Compared with those in healthy people at high altitude, some health indicators such as BMI, leukocyte count, neutrophil count, lymphocyte count, platelet count, systolic blood pressure, oxygen saturation, LVEF value and LVFS value were lower but other indicators including the blood viscosity, hematocrit, hemoglobin concentration and heart rate were higher in patients with HADT. CONCLUSIONS: The prevalence rate of HADT (66.9%) was high among people moving to a high altitude of 4516 m. Clinical characteristics of HADT were: (1) Impairment of left ventricular systolic function; (2) Immune depression; (3) Microcirculation disturbance; and (4) Decline of hemostasis and coagulation function.

Retina Is Protected by Neuroserpin from Ischemic/Reperfusion-Induced Injury Independent of Tissue-Type Plasminogen Activator.

The purpose of the present study was to investigate the potential neuroprotective effect of neuroserpin (NSP) on acute retinal ischemic/reperfusion-induced (IR) injury. An IR injury model was established by elevating intraocular pressure (IOP) for 60 minutes in wild type and tPA-deficient (tPA-/-) mice. Prior to IR injury, 1 µL of 20 µmol/L NSP or an equal volume of bovine serum albumin (BSA) was intravitreally administered. Retinal function was evaluated by electroretinograph (ERG) and the number of apoptotic neurons was determined via TUNEL labeling. Caspase-3, -8, -9,poly (ADP-ribose) polymerase (PARP)and their cleaved forms were subsequently analyzed. It was found that IR injury significantly damaged retinal function, inducing apoptosis in the retina, while NSP attenuated the loss of retinal function and significantly reduced the number of apoptotic neurons in both wild type and tPA-/- mice. The levels of cleaved caspase-3, cleaved PARP (the substrate of caspase-3) and caspase-9 (the modulator of the caspase-3), which had increased following IR injury, were significantly inhibited by NSP in both wild type and tPA-/- mice. NSP increased ischemic tolerance in the retina at least partially by inhibiting the intrinsic cell death signaling pathway of caspase-3. It was therefore concluded that the protective effect of neuroserpin maybe independent from its canonical interaction with a tissue-type plasminogen activator.

Two triterpeniods from Cyclocarya paliurus (Batal) Iljinsk (Juglandaceae) promote glucose uptake in 3T3-L1 adipocytes: The relationship to AMPK activation.

PURPOSE: The current study investigated the efficacy of Cyclocarya paliurus chloroform extract (CPEC) and its two specific triterpenoids (cyclocaric acid B and cyclocarioside H) on the regulation of glucose disposal and the underlying mechanisms in 3T3-L1 adipocytes. METHODS: Mice and adipocytes were stimulated by macrophages-derived conditioned medium (Mac-CM) to induce insulin resistance. CPEC was evaluated in mice for its ability by oral glucose tolerance test (OGTT) and insulin tolerance test (ITT). To investigate the hypoglycemic mechanisms of CPEC and its two triterpenoids, glucose uptake, AMP-activated protein kinase (AMPK) activation, inhibitor of NF-κB kinase ß (IKKß) phosphorylation and insulin signaling transduction were detected in 3T3-L1 adipocytes using 2-NBDG uptake assay and Western blot analysis. RESULTS: Mac-CM, an inflammatory stimulus which induced the glucose and insulin intolerance, increased phosphorylation of IKKß, reduced glucose uptake and impaired insulin sensitivity. CPEC and two triterpenoids improved glucose consumption and increased AMPK phosphorylation under basal and inflammatory conditions. Moreover, CPEC and its two triterpenoids not only enhanced glucose uptake in an insulin-independent manner, but also restored insulin-mediated protein kinase B (Akt) phosphorylation by reducing the activation of IKKß and regulating insulin receptor substrate-1 (IRS-1) serine/tyrosine phosphorylation. These beneficial effects were attenuated by AMPK inhibitor compound C, implying that the effects may be associated with AMPK activation. CONCLUSIONS: CPEC and its two triterpenoids promoted glucose uptake in the absence of insulin, as well as ameliorated IRS-1/PI3K/Akt pathway by inhibiting inflammation. These effects were related to the regulation of AMPK activity.

Psychological and cognitive impairment of long-term migrators to high altitudes and the relationship to physiological and biochemical changes.

BACKGROUND AND PURPOSE: The present study aimed to examine how long-term migration to high-altitude regions affects mentality and cognition, and the correlation with various physiological and biochemical changes. METHODS: The WHO Neurobehavioral Core Test Battery, Raven's Standard Progressive Matrices (RSPM) and Pittsburgh Sleep Quality Index questionnaire were used to assess 141 young male subjects who lived in plain regions and 217 young male subjects who had migrated to a 4500 m high-altitude region and lived there for 1-5 years. Arterial oxyhemoglobin saturation, cerebral tissue oxygenation indices (TOIs), serum S100B and brain-derived neurotrophic factor (BDNF) were also measured. RESULTS: Long-term migrators to a high-altitude region exhibited exacerbated mood disorders, retarded color discrimination ability, decreased visual memory capacity, and impaired perceptual motor skill and motion stability. In addition, the migrators exhibited lower RSPM scores and lower sleep quality. Further analyses revealed significant correlations between sleep quality and cerebral TOIs, mood and sleep quality, mood and certain cognitive functions, mood and serum BDNF levels, and RSPM scores and serum S100B levels. CONCLUSIONS: Long-term living at high altitudes causes significant impairment of psychological and cognitive function. Cerebral hypoxic extent, sleep quality and biochemical dysfunction are major influencing factors.

Opposite effects of genistein on the regulation of insulin-mediated glucose homeostasis in adipose tissue.

BACKGROUND AND PURPOSE: Genistein is an isoflavone phytoestrogen found in a number of plants such as soybeans and there is accumulating evidence that it has beneficial effects on the regulation of glucose homeostasis. In this study we evaluated the effect of genistein on glucose homeostasis and its underlying mechanisms in normal and insulin-resistant conditions. EXPERIMENTAL APPROACH: To induce insulin resistance, mice or differentiated 3T3-L1 adipocytes were treated with macrophage-derived conditioned medium. A glucose tolerance test was used to investigate the effect of genistein. Insulin signalling activation, glucose transporter-4 (GLUT4) translocation and AMP-activated PK (AMPK) activation were detected by Western blot analysis or elisa. KEY RESULTS: Genistein impaired glucose tolerance and attenuated insulin sensitivity in normal mice by inhibiting the insulin-induced phosphorylation of insulin receptor substrate-1 (IRS1) at tyrosine residues, leading to inhibition of insulin-mediated GLUT4 translocation in adipocytes. Mac-CM, an inflammatory stimulus induced glucose intolerance accompanied by impaired insulin sensitivity; genistein reversed these changes by restoring the disturbed IRS1 function, leading to an improvement in GLUT4 translocation. In addition, genistein increased AMPK activity under both normal and inflammatory conditions; this was shown to contribute to the anti-inflammatory effect of genistein, which leads to an improvement in insulin signalling and the amelioration of insulin resistance. CONCLUSION AND IMPLICATIONS: Genistein showed opposite effects on insulin sensitivity under normal and inflammatory conditions in adipose tissue and this action was derived from its negative or positive regulation of IRS1 function. Its up-regulation of AMPK activity contributes to the inhibition of inflammation implicated in insulin resistance.

Naloxone inhibition of lipopolysaccharide-induced activation of retinal microglia is partly mediated via the p38 mitogen activated protein kinase signalling pathway.

OBJECTIVES: To investigate the effects and underlying mechanism of action of naloxone on lipopolysaccharide (LPS)-induced activation of retinal microglia in vitro. METHODS: Rat retinal microglia primary cultures were divided into four treatment groups: untreated; 1 µg/ml LPS for 12 h; 0.5, 1.0 or 2.0 µM naloxone for 30 min before LPS; 2.5 or 5.0 µM SB203580 for 12 h before LPS and naloxone. Levels of tumour necrosis factor (TNF)-α and interleukin (IL)-1ß were determined by enzyme-linked immuno sorbent assay. Western blot analysis and double immunofluorescence were used to examine activation of the mitogen activated protein kinase (MAPK) signalling pathway. RESULTS: LPS induced an increase in TNF-α and IL-1ß in culture supernatants, which was dose-dependently inhibited by naloxone. Naloxone also dose-dependently inhibited LPS-induced increases in phosphorylated p38 MAPK. Pretreatment of cells with SB203580 attenuated the inhibitory effect of naloxone on TNF-α and IL-1ß production. CONCLUSIONS: Naloxone suppressed LPS-induced activation of cultured retinal microglia and this suppression appeared to occur partly through the p38 MAPK signalling pathway. Naloxone may have therapeutic potential in neuro degenerative diseases characterized by the activation of microglia.

A novel single-base deletion mutation of the RUNX2 gene in a Chinese family with cleidocranial dysplasia.

We identified a disease-causing mutation of the RUNX2 gene in a four-generation Chinese family affected with cleidocranial dysplasia (CCD). For mutation analysis, the coding region of RUNX2 was sequenced with DNA from two patients and three unaffected family members. The RUNX2 mutation was investigated in 50 normal controls by denaturing high pressure liquid chromatography. A heterozygous single-base deletion (c.549delC) of RUNX2, which predicts a termination site at the 185th codon and leads to a stop in the runt domain of RUNX2 protein, was detected in both patients but not in the three unaffected members of the family. This mutation was also not found in 50 controls and has not been reported previously. We demonstrated that a novel mutation (c.549delC) of RUNX2 is associated with CCD in a Chinese family, adding to the repertoire of RUNX2 mutations related to CCD.

Estrogen attenuates VEGF-initiated blood-retina barrier breakdown in male rats.

17ß-Estradiol has been demonstrated to protect blood-brain barrier from disruption and attenuate brain injury in various conditions. The aim of this study was to investigate the effect of 17ß-estradiol on the blood-retina barrier (BRB) breakdown induced by intravitreous injection of vascular endothelial growth factor (VEGF), a significant mediator of vascular permeability. Intravitreous injection of VEGF was performed to initiate BRB breakdown in male rats with PBS in the contralateral eye as control. 2 doses of 17ß-estradiol and vehicle control were given to 3 groups of rats. The integrity of the BRB was quantified by Evans blue technique and assessed by fluorescent dyes in retinal sections and wholemounts. BRB breakdown was achieved by VEGF as retinal vascular permeability was increased compared with control eyes (14.66±4.09 vs. 4.94±1.20 µl/g/h, p<0.01). Vascular permeability in the 2 groups treated with 17ß-estradiol was reduced compared with control (14.66±4.09 vs. 10.26±3.67 vs. 7.37±2.22 µl/g/h, p<0.01). Rhodamine isothiocyanate (RhIC) extravasation in retinal sections and Evans blue-albumin complex leakage in retinal wholemounts were also decreased in the 2 treatment groups. These results suggest that 17ß-estradiol attenuates BRB breakdown induced by VEGF in male rats, which may provide a new role of 17ß-estradiol in ocular diseases.