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Background. Resting-state functional magnetic resonance imaging (rs-fMRI) is a promising method for the study of brain function. Typically, rs-fMRI is performed on anesthetized animals. Although different functional connectivity (FC) in various anesthetics on whole brain have been studied, few studies have focused on different FC in the aged brain. Here, we measured FC under three commonly used anesthesia methods and analyzed data to determine if the FC in whole brain analysis were similar among groups. Methods. Twenty-four male aged Wistar rats were randomly divided into three groups (n = 8 in each group). Anesthesia was performed under either isoflurane (ISO), combined ISO + dexmedetomidine (DEX) or α-chloralose (AC) according to the groups. Data of rs-fMRI was analyzed by FC in a voxel-wise way. Differences in the FC maps between the groups were analyzed by one-way analysis of variance and post hoc two-sample t tests. Results. Compared with ISO + DEX anesthesia, ISO anesthesia caused increased FC in posterior brain and decreased FC in the middle brain of the aged rat. AC anesthesia caused global suppression as no increase in FC was observed. Conclusion. ISO could be used as a substitute for ISO + DEX in rat default mode network studies if the left temporal association cortex is not considered important.

BACKGROUND The aim of this study was to analyze the value of ocular endoscopy in detecting and extracting intraocular cilia in patients with ocular trauma. MATERIAL AND METHODS We retrospectively analyzed data on identification and extraction of 46 intraocular cilia in 16 eyes with open-globe injury during endoscope-assisted vitrectomy. RESULTS A total of the 16 patients with open-globe injury were operated on from September 2002 to June 2019. The cornea in 14 eyes was cloudy. Two eyes had endophthalmitis and 13 eyes had retinal detachment. A total of 46 cilia were extracted through direct observation under the ocular endoscope during vitrectomy 1 to 68 weeks after injury. The number of cilia per eye varied from 1 to 10. Most of the cilia were located in or near the wound. Postoperative IOP was normal in 14 patients. The follow-up after surgery showed hypotony in only 2 eyes (7.2 and 5.8 mmHg, respectively). Compared with preoperative intraocular pressure, there was a statistically significant difference. The postoperative visual acuity improved in 12 eyes and remained unchanged in 3 eyes. The vision after surgery was significantly improved compared with that before surgery (P=0.006). The intraocular pressure increased significantly after operation (P0.001). And no glaucoma or retinal detachment or endophthalmitis was found. No eyes needed additional vitreous surgery. CONCLUSIONS Ocular endoscopy allows surgeons to detect intraocular cilia that were no undetected by CT or B-ultrasound preoperatively in time and to extract them effectively. It improves performance of vitrectomy in the presence of a cloudy cornea and also prevents exogenous endophthalmitis. The vision of patients with ocular trauma was improved.

BACKGROUND Endophthalmitis, or inflammation of the internal tissues of the eye, may follow trauma and results in loss of vision if not treated promptly. Vitrectomy is used to debride the inflamed vitreous. This retrospective study aimed to compare the outcome from conventional vitrectomy (CV) and endoscope-assisted vitrectomy (EAV) performed for endophthalmitis within five days and 10 days after open ocular trauma. MATERIAL AND METHODS The clinical data from 18 patients (18 eyes) with endophthalmitis caused by open ocular trauma were analyzed at a single center. CV or EAV was performed based on the degree of vision and the involvement of the vitreous cavity. RESULTS Of the 18 patients with endophthalmitis, seven patients underwent EAV, and 11 patients underwent CV. The number of cases with preoperative corneal laceration, edema, bleeding, and formation of corneal nebular opacity was higher in the EAV group compared to the CV group. Four patients had retinal detachment, of which three patients were treated with EAV. Seventeen patients had their sight restored by a single vitrectomy procedure. In 10 cases, the culture of the vitreous fluid was positive for infection, and eight patients had Gram-positive Staphylococcus epidermidis infection. Follow-up showed that vitrectomy performed within five days (P=0.001) and 10 days (P=0.047) of open ocular trauma resulted in significant improvement of visual acuity. CONCLUSIONS Vitrectomy for endophthalmitis due to open ocular trauma performed within five days of injury restored visual acuity. EAV was shown to be an effective alternative to CV.

Background Study shows that signal transducer and activator of transcription 3 (STAT3) can increase the Warburg effect by stimulating hexokinase 2 in breast cancer and upregulate lactate dehydrogenase A and pyruvate dehydrogenase kinase 1 in myeloma. STAT3 and pyruvate kinase M2 (PKM2) can also be activated and enhance the Warburg effect in hepatocellular carcinoma. Precancerous lesions are critical to human and rodent hepatocarcinogenesis. However, the underlying molecular mechanism for the development of liver precancerous lesions remains unknown. We hypothesized that STAT3 promotes the Warburg effect possibly by upregulating p-PKM2 in liver precancerous lesions in rats. Aim To investigate the mechanism of the Warburg effect in liver precancerous lesions in rats. Methods A model of liver precancerous lesions was established by a modified Solt-Farber method. The liver pathological changes were observed by HE staining and immunohistochemistry. The transformation of WB-F344 cells induced with N-methyl-N'-nitro-N-nitrosoguanidine and hydrogen peroxide was evaluated by the soft agar assay and aneuploidy. The levels of glucose and lactate in the tissue and culture medium were detected with a spectrophotometer. The protein levels of glutathione S-transferase-π, proliferating cell nuclear antigen (PCNA), STAT3, and PKM2 were examined by Western blot and immunofluorescence. Results We found that the Warburg effect was increased in liver precancerous lesions in rats. PKM2 and p-STAT3 were upregulated in activated oval cells in liver precancerous lesions in rats. The Warburg effect, p-PKM2, and p-STAT3 expression were also increased in transformed WB-F344 cells. STAT3 activation promoted the clonal formation rate, aneuploidy, alpha-fetoprotein expression, PCNA expression, G1/S phase transition, the Warburg effect, PKM2 phosphorylation, and nuclear translocation in transformed WB-F344 cells. Moreover, the Warburg effect was inhibited by stattic, a specific inhibitor of STAT3, and further reduced in transformed WB-F344 cells after the intervention for PKM2. Conclusion The Warburg effect is initiated in liver precancerous lesions in rats. STAT3 activation promotes the Warburg effect by enhancing the phosphorylation of PKM2 in transformed WB-F344 cells.

Purpose This study examined the role of the CSF1/CSF1Raxis in the crosstalk between choroidal vascular endothelial cells (CVECs) and macrophages during the formation of choroidal neovascularization (CNV). Methods Quantitative reverse transcriptase (QRT)-PCR, Western blot and ELISA measured the production and release of CSF1 from human choroidal vascular endothelial cells (HCVECs) under hypoxic conditions. Western blot detected CSF1 released from HCVECs under hypoxic conditions that activated the PI3K/AKT/FOXO1 axis in human macrophages via binding to CSF1R. Transwell migration assay, qRT-PCR, and Western blot detected the effect of CSF1 released from HCVECs on macrophage migration and M2 polarization via the CSF1R/PI3K/AKT/FOXO1 pathway. Incorporation of 5-ethynyl-20-deoxyuridine, transwell migration, and tube formation assays detected the effects of CSF1/CSF1R on the behaviors of HCVECs. Fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA), and immunofluorescence detected the effect of blockade of CSF1/CSF1R on mouse laser-induced CNV. Color fundus photograph, ICGA, and FFA detected CNV lesions in neovascular AMD (nAMD) patients. ELISA detected CSF1 and CSF1R in the aqueous humor of age-related cataract and nAMD patients. Results CSF1 released from HCVECs under hypoxic conditions activated the PI3K/AKT/FOXO1 axis in human macrophages via binding to CSF1R, promoting macrophage migration and M2 polarization via up-regulation of the CSF1R/PI3K/AKT/FOXO1 pathway. Human macrophages promoted the proliferation, migration, and tube formation of HCVECs in a CSF1/CSFR1-dependent manner under hypoxic conditions. CSF1/CSF1R blockade ameliorated the formation of mouse laser-induced CNV. CSF1 and CSF1R were increased in the aqueous humor of nAMD patients. Conclusions Our results affirmed the crucial role of CSF1/CSF1R in boosting the formation of CNV and offered potential molecular targets for the treatment of nAMD.

BACKGROUND This study aimed to investigate the effect of cobalt transporter II gene (rs1801198, rs2301957, rs9606756) polymorphism on serum homocysteine level and its correlation with young and middle recurrent cerebral infarction. METHODS A total of 348 young and middle-aged patients with cerebral infarction admitted to The Third Affiliated Hospital of Qiqihar Medical University from January 2017 to March 2018 were enrolled. The patients were divided into recurrent and non-recurrent groups according to follow-up. DNA was extracted from the peripheral blood of patients, and the DNA samples were genotyped by IlluminaBeadArray technology to detect the gene polymorphisms of cobalt transporter II (TCN2) sites (rs1801198, rs2301957, rs9606756), and the homocysteine (hcy) level was determined by cyclic enzymatic method. VitB12 and folate levels were measured by chemiluminescence immunoassay, and holo transcobalamin (holoTC) expression levels were detected by enzyme-linked immunosorbent assay. RESULTS The frequency of alleles of rs9606756 mutation in the recurrent group was higher than that in the non-recurrent group (P

Choroidal neovascularization (CNV), a feature of neovasular age-related macular degeneration (AMD), acts as a leading cause of vision loss in the elderly. Shikonin (SHI), a natural bioactive compound extracted from Chinese herb radix arnebiae, exerts anti-inflammatory and anti-angiogenic roles and also acts as a potential pyruvate kinase M2 (PKM2) inhibitor in macrophages. The major immune cells macrophages infiltrate the CNV lesions, where the production of pro-angiognic cytokines from macrophage facilitates the development of CNV. PKM2 contributes to the neovascular diseases. In this study, we found that SHI oral gavage alleviated the leakage, area and volume of mouse laser-induced CNV lesion and inhibited macrophage infiltration without ocular cytotoxicity. Moreover, SHI inhibited the secretion of pro-angiogenic cytokine, including basic fibroblast growth factor (FGF2), insulin-like growth factor-1 (IGF1), chemokine (C-C motif) ligand 2 (CCL2), placental growth factor and vascular endothelial growth factor (VEGF), from primary human macrophages by down-regulating PKM2/STAT3/CD163 pathway, indicating a novel potential therapy strategy for CNV.

Testicular germ cell tumors (TGCT) are the most common cancer among men aged 15 to 39 years. Previous studies have considered factors related to TGCT survival rate and race/ethnicity, but histological type of the diagnosed cancer has not yet been thoroughly assessed.The data came from 42,854 eligible patients from 1992 to 2015 in the Surveillance Epidemiology and End Results 18. Frequencies and column percent by seminoma and nonseminoma subtypes were determined for each covariates. We used Cox proportional hazard regression to assess the impact of multiple factors on post-diagnostic mortality of TGCT.Black males were diagnosed at a later stage, more commonly with local or distant metastases. The incidence of TGCT in black non-seminoma tumors increased most significantly. The difference in survival rates between different ethnic and histological subtypes, overall survival (OS) in patients with non-seminoma was significantly worse than in patients with seminoma. The most important quantitative predictor of death was the stage at the time of diagnosis, and older diagnostic age is also important factor affecting mortality.Histological type of testicular germ cell tumor is an important factor in determining the prognosis of testicular cancer in males of different ethnic groups.

Recently, it has been demonstrated that microRNA-137 (miR-137) plays a vital role in the induction of oxidative stress of neurons in Parkinson’s disease (PD). Herein, the study aimed to investigate the effects of serum exosomal miR-137 on oxidative stress injury of neurons in PD. Microarray analysis was adopted to screen the PD-related differential expressed genes and predict the interaction between OXR1 and miR-137 in PD. It was found that OXR1 was down-regulated while miR-137 was up-regulated in PD. Additionally, miR-137 targeted OXR1 and negatively regulated its expression. Mouse and neuron models of PD were established to mimic the pathological changes, especially oxidative stress injury induced by PD. The significance of miR-137 and OXR1 in oxidative stress injury was investigated in neuron model of PD using gain- and loss-of-function approaches. The obtained data exhibited that inhibition of miR-137 or up-regulation of OXR1 ameliorated PD-induced oxidative stress injury, reduced pole-climbing time, but increased score for traction test as well as promoted viability and decreased apoptosis of neurons in PD model, accompanied with decreased MDA content and ROS levels, and increased SOD levels. Furthermore, PD mice were injected with serum-derived exosomes or neurons in PD models were exposed to exosomes derived from serum of PD mice. Loss-of-function experiments using miR-137 antagomir exhibited that inhibition of exosomal miR-137 ameliorated PD-induced oxidative stress injury in vitro, reduced pole-climbing time but increased score for traction test in vivo. Collectively, down-regulation of exosomal miR-137 alleviates oxidative stress injury in PD by up-regulating OXR1.

Age-related macular degeneration (AMD) is a leading cause of vision loss in the elderly and is attributed to choroidal neovascularization (CNV), which is a feature of wet AMD. The hypoxia-inducible factor-1α (HIF-1α)/vascular endothelial growth factor (VEGF)/VEGF receptor 2 (VEGFR2) pathway contributes to the pathogenesis of CNV. M1-type macrophages/microglia secrete interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α), facilitating the development of CNV. Epigallocatechin-3-gallate (EGCG) is a kind of polyphenol in green tea that exerts anti-inflammatory and antiangiogenic effects. In this study, a prodrug of EGCG (pro-EGCG) alleviated mouse laser-induced CNV leakage and reduced CNV area by down-regulating HIF-1α/VEGF/VEGFR2 pathway; M1-type macrophage/microglia polarization; as well as endothelial cell viability, proliferation, migration and tube formation, indicating a novel potential therapy for AMD.

Aims We investigated the protective effects of chemical stimulation of cerebellar fastigial nucleus (FN) on stress gastric mucosal injury (SGMI) and its possible neuro-regulatory mechanisms in rats. Main methods Chemical stimulation, electrical stimulation, chemical ablation, electrolytic lesion, and microinjection were used to investigate the effects of FN simulation on SGMI. The model of SGMI was established by restraint and water (21 ± 1 °C)-immersion (RWI) for 3 h in rats. The gastric mucosal injury index indicated the severity of gastric mucosal injuries. Key findings We showed that microinjection of L-glutamic acid into the FN or electrical stimulation of the FN markedly attenuated SGMI. Either chemical lesion of the FN or electrical ablation of the decussation of superior cerebellar peduncle (DSCP) obviously aggravated SGMI. The protective effect of FN stimulation on SGMI was reversed after chemical ablation of the lateral hypothalamic area (LHA). The protective effect of FN was prevented by pretreatment with the glutamic acid decarboxylase antagonist, 3-MPA into the FN or GABAA receptor antagonist, bicuculline into the LHA. The protective effect of FN was abolished by pretreatment with sympathectomy. The discharge frequency of greater splanchnic nerve (GSN) was decreased and gastric mucosal blood flow (GMBF) was increased after chemical stimulation of FN. These results indicate that the FN participates in regulation of SGMI, and is a specific area in the CNS for exerting protective effects on the SGMI. The DSCP, LHA and peripheral sympathetic nerve may be involved in this process. Significance Our findings might provide a new and improved understanding of the cerebellar function and an effective treatment strategy for stress gastric mucosal injury.

Aims To investigate the protective effects of 17β-estradiol postconditioning against ischemia/reperfusion (I–R)-induced gastric mucosal injury in rats. Main methods The animal model of gastric ischemia/reperfusion was established by clamping of the celiac artery for 30 min and reperfusion for 30 min, 1 h, 3 h, 6 h, 12 h or 24 h. 17β-estradiol at doses of 5, 50 or 100 μg/kg (rat) was administered via peripheral veins 2 min before reperfusion. In a subgroup of rats, the estrogen receptor antagonist fulvestrant (Ful, 2 mg/kg) was intravenously injected prior to 17β-estradiol administration. Histological and immunohistochemical methods were employed to assess the gastric mucosal injury index and gastric mucosal cell apoptosis and proliferation. The malondialdehyde (MDA) concentration, superoxide dismutase (SOD) activity, xanthine oxidase (XOD) activity and hydroxyl free radical (–OH) inhibitory ability were determined by colorimetric assays. Subsequently, the expression of Bcl-2 and Bax in rat gastric mucosa was examined by western blotting. Key findings 17β-estradiol dose-dependently inhibited gastric I–R (GI–R) injury, and 17β-estradiol (50 μg/kg) markedly attenuated GI–R injury 1 h after reperfusion. 17β-estradiol inhibited gastric mucosal cell apoptosis and promoted gastric mucosal cell proliferation in addition to increasing SOD activity and –OH inhibitory ability and decreasing the MDA content and XOD activity. The Bax protein level increased 1 h after GI–R and was markedly reduced by intravenous administration of 17β-estradiol. In contrast, the level of Bcl-2 protein decreased 1 h after GI–R and was restored to normal levels by intravenous administration of 17β-estradiol. These effects of 17β-estradiol were inhibited by pretreatment with fulvestrant. Significance 17β-estradiol postconditioning should be investigated further as a possible strategy against gastric mucosal injury.

A systematic study on the structure and function of Glucose-6-phosphate dehydrogenase (G6PD) variations was carried out in China. A total of 155,879 participants were screened for G6PD deficiency by the G6PD/6PGD ratio method and 6,683 cases have been found. The prevalence of G6PD deficiency ranged from 0 to 17.4%. With informed consent, 1,004 cases from 11 ethnic-based groups were subjected to molecular analysis. Our results showed the followings: (1) The G6PD variants are consistent across traditional ethnic boundaries, but vary in frequencies across ethnic-based groups in Chinese population, (2) The G6PD variants in Chinese population are different from those in African, European, and Indian populations, (3) A novel G6PD-deficiency mutation, 274C--T, has been found, and (4) Denaturing high performance liquid chromatography is of great advantage to detecting G6PD-deficient mutations for diagnosis and genetic counseling. Moreover, functional analysis of the human G6PD variants showed the following: (1) The charge property, polarity, pK-radical and side-chain radical of the substituting amino acid have an effect on G6PD activity, (2) The G6PDArg459 and Arg463 play important roles in anchoring NADP+ to the catalytic domain to maintain the enzymatic activity, and (3) The sequence from codon 459 to the carboxyl terminal is essential for the enzymatic function.

Aging results in progressive glomerular sclerosis and reductions in glomerular filtration rate (GFR). Oxidative stress may be an important mechanism for the aging process, but to date the role of oxidative stress on renal aging has not been determined. The present study was performed to determine whether age-related alterations in renal hemodynamics and morphology were associated with oxidative stress and whether this could be attenuated by chronic administration of vitamin E. Rats, aged 13 mo, were given either control diet containing vitamin E 50 IU/kg ( n = 6) or a high-vitamin E diet (5,000 IU/kg; n = 6) for 9 mo. Another group of rats (3–4 mo old; n = 7) served as young controls. Aging was accompanied by a 60% reduction in GFR, a threefold increase in renal F2isoprostanes, newly discovered vasoconstrictive F2-like prostaglandins generated by free radical-mediated lipid peroxidation. Renal aging was also associated with an increase in oxidant-sensitive heme oxygenase, advanced glycosylation end products (AGEs), and the AGE receptor, RAGE. AGE-RAGE interaction has been shown to induce oxidative stress. With high-vitamin E diet, GFR was increased by 50%, F2isoprostanes were suppressed, and expression of heme oxygenase and RAGE was attenuated. There was also a tendency for glomerular sclerosis to be attenuated. These data demonstrate that age-related decline in renal function is accompanied by oxidative stress and that administration of antioxidants, such as vitamin E, could attenuate the decline in renal function.

We report the genetic data of 435 patients with Hb H (β4) disease who presented at our center between 2005 and 2012. Our results showed that all patients had the Southeast Asian deletion (- -(SEA)) on one allele. The -α(3.7) (rightward) deletion was the most common on the other allele, followed by the -α(4.2) (leftward) deletion, Hb Constant Spring (Hb CS, α142, Term → Gln; HBA2: c.427T C) and Hb Quong Sze [Hb QS, α125(H8)Leu → Pro; HBA2: c.377T C] mutations. Two rare point mutations, α31(B12)Arg → Lys; HBA2: c.95GA and Hb Zurich Albisrieden [α59(E8)Gly → Arg; HBA1: c.178G C], were also identified. Four patients had a concomitant β-thalassemia (β-thal) heterozygosity. Our results reflect the genetic heterogeneity of Hb H disease and the interaction between Hb H disease and β-thal trait in Southern China.

To study the feasibility and effectiveness of the extraction of intraocular foreign body (IOFB) at or near the ciliary body by using endoscopic vitrectomy.It was a retrospective case series study. Clinical materials and data of 15 patients (16 eyes) with IOFB at or near the ciliary body from July 2002 to December 2010 were analyzed retrospectively. Vitrectomy and IOFB extraction under endoscope were conducted in all cases.Thirty six IOFB (18 magnetic and 18 nonmagnetic IOFBs) were extracted successfully. The size of IOFB ranged from smaller than 1 mm×1 mm×1 mm to 9 mm×6 mm×4 mm. Among them, 31 IOFB were extracted from ciliary body or nearby and 5 IOFB were located at other places. The cornea was opaque before the operation in 11 eyes. Postoperative visual acuity was improved in 12 eyes. Vision was improved from finger-counting and below to 0.1-0.3 in 4 eyes. Intraocular pressure was normal in 14 eyes. The retina was reattached in 12 eyes. The cornea was clearer than before in 8 eyes. After the end of the fellow-up, vision was remained in finger-counting and below in only 7 eyes.There was almost no blind spot by using endoscope to observe the ciliary body, periphery retina tear or other pathological changes, the IOFB at or near ciliary body and the whole retina in the vitrectomy. This makes the extraction of IOFB at or near the ciliary body and the conduct of cyclophotocoagulation more accurate and simple.

1. Nitric oxide (NO), a gas transmitter, modulates many physiological processes, including the central regulation of cardiovascular activity. However, the mechanisms underlying the regulation of cardiovascular activity remain relatively unexplored. In the present study, we hypothesized that central NO-dependent sympathetic inhibition is mediated by activation of adenosine A(2A) receptors (A(2A)R) and inhibition of acetylcholine (ACh) release in the rostral ventrolateral medulla (RVLM). 2. L-Arginine (L-Arg; an NO donor; 100 nmol/100 nL) was microinjected into the RVLM of male Sprague-Dawley rats and heart rate variability (HRV) was assessed as an index of cardiac sympathovagal balance. Following microinjection of L-Arg, decreases were seen in mean arterial pressure (MAP), heart rate (HR) and the ratio of the low- to high-frequency components (LF/HF) of HRV. Pretreatment of rats with SCH58261 (40 pmol/60 nL into the RVLM), a competitive antagonist of the A(2A) R, attenuated these effects. 3. Western blot analysis and ELISA revealed that adenosine and A(2A)R levels increased in the RVLM following L-Arg microinjection, whereas ACh and muscarinic M(1) receptor levels decreased significantly, in parallel with the cardiovascular responses to L-Arg microinjection. The decrease in ACh levels was abolished by SCH58261 pretreatment. 4. Microinjection of N(G)-nitro-L-arginine methyl ester (a non-selective inhibitor of NO synthase; 15 nmol/100 nL) into the RVLM significantly increased MAP, HR and sympathetic activity, as evidenced by HRV (LF, HF and the LF/HF ratio were all increased). 5. The results indicate that the central NO/NO synthase system in the RVLM may modulate cardiovascular activity by activating the A(2A)R, which subsequently inhibits activation of the muscarinic M(1) receptor.

The present study aimed to investigate the protective mechanism of N-acetylcysteine (NAC) against gastric ischemia /reperfusion (GI/R) injury in rats. After intravenous injection (IV) of NAC (150 mg/kg) into femoral vein, the rats were subjected to 30 min of ischemia induced by clamping the celiac artery followed by 60 min of reperfusion. After the gastric mucosal damage index (GMDI) had been calculated, gastric mucosal cell in situ apoptosis was detected by TUNEL method. The protein expression of p-ERK, p-JNK and NF-kappaB, and mRNA expression of TNF-alpha and Caspase-3 in gastric mucosa were evaluated by using Western-blot or RT-PCR, respectively. The results showed that NAC not only attenuated the GI-R injury, but also decreased gastric mucosal cellular apoptosis. Furthermore, NAC increased the protein expression of p-ERK, while inhibited protein expression of p-JNK, NF-kappaB in gastric mucosa. NAC also decreased the expression of TNF-alpha mRNA and Caspase-3 mRNA in gastric mucosa. Capsazepine (CPZ) (400 mg/kg, IV) reversed the protective effect of NAC against GI/R injury in rats. These results suggest that NAC can protect rats against GI/R injury. This protective effect is possibly mediated by the up-regulation of p-ERK and down-regulation of p-JNK and NF-kappaB. In addition, vanilloid receptor subtype 1 may be involved in the protective mechanism of NAC against GI/R injury.

To identify mutations of iduronate-2-sulfatase (IDS) gene in a patient with Hunter syndrome and to establish a basis for prenatal gene diagnosis of Hunter syndrome. Urine GAG assay was used for preliminary diagnosis of mucopolysaccharidosis. PCR from dried blood spots and DNA sequencing were applied to analyze hot spot mutations in exons 9, 3, 8 of the IDS gene in the proband and his parents. A new missense mutation (T1140C) in exon 8 of the IDS gene was found by DNA sequencing. This mutation caused a substitution of codon 339 from CTA (leucine) to CCA (proline). The patient is a hemizygote and his mother is a heterozygote. The new missense mutation results in a change in the primary and tertiary structure of the IDS protein. It is possible that this mutation severely impairs the enzymatic activity and is the underlying basis for the pathology seen in this patient with Hunter syndrome.

Mutations of the iduronate-2-sulfatase (IDS) gene is the ultimate cause of Hunter syndrome. Clarification of the nature of mutations will create a necessary premise for prenatal gene diagnosis. A mucopolysaccharidosis (MPS) type II patient and his parents from an ethnic minority in Yunnan province were studied to identify their possible mutation in IDS gene to establish the basis for prenatal gene diagnosis.The patient was a boy, 6 years and 10 months old. Urine glycosaminoglycans (GAGs) assay was used for preliminary diagnosis of the patient and his parents with the disease. The three related persons' DNA was extracted and the concentration and purity of the DNA were measured after the urine test results confirmed the diagnosis. Polymerase chain reaction-denaturing high performance liquid chromatography (PCR-DHPLC) analysis was performed to detect the position of the mutation around the hot spots of mutation in exon 9, 3, 8 of the IDS gene. DNA bidirectional direct sequencing was applied to analyze the mutation detected by PCR-DHPLC.The results of GAGs test showed that in the child with MPS, dermatan sulfate (DS) was positive (+++), heparan sulfate (HS) (+++), chondroitin sulfate (CS) and keratan sulfate (KS) were negative (-); while in his parents none of DS, HS, CS and KS was positive. Abnormal peaks in exon 9 of IDS gene shown by PCR-DHPLC were found in the patient. His mother had heterozygotic peaks. A new frame-mutation (1343-TT) in exon 9 of IDS gene of this patient was confirmed by DNA sequencing. The position where mutation occurred was inside codon 407 (TTT), that means two "T" deleted at position 1343 base pair (1343-TT) in cDNA of the IDS gene, caused a new frame-mutation. It caused elongation of the amino acid chain to a terminal codon TGA at position 429. Thus the peptide chain was shortened from 550 to 428 amino acids. The patient is a hemizygote of the mutation and his mother is a heterozygote.A new frame-mutation (1343-TT) on the IDS gene was identified in this study. The patient is a hemizygote and his mother is a heterozygote. The mutation (1343-TT) resulted in loss of 122 amino acids, which probably caused seriously decreased enzyme activity of IDS, and the authors speculate that this mutation may be the pathological basis of the disease. So, if the mother becomes pregnant again, a prenatal gene diagnostic test for the same mutation should be performed. Furthermore, PCR-DHPLC followed by DNA sequencing are effective methods for diagnosis, including prenatal diagnosis of MPS II.

To identify the mutations of iduronate-2-sulfatase (IDS) gene, and to establish a basis of prenatal gene diagnosis of Hunter syndrome.Urine glycosaminoglycan (GAG) assay was used to preliminary diagnosis of mucopolysaccharidosis. PCR-denaturing high-performance liquidchromatograptly (PCR-DHPLC) analysis was performed to detect the mutation in exons 9, 3, 8 of the IDS gene. DNA sequencing was applied to analyze the mutation detected by PCR-DHPLC.Abnormal peaks were found by PCR-DHPLC. A new frame-mutation (1569+TT) in exon 9 of IDS gene was identified by DNA sequencing. Two "T"q inserted in position 1569 base pair (1569+TT) caused a substitution of codon 482 (TTA, leucine) to 482 (TTT, phenylalanine). The "TT" insertion results in the decrease of amino acids from 550 to 482. The patient is a hemizygote and his mother is a heterozygote.A new frame-shift mutation of IDS gene is found to report. The mutation (1569+TT) results in 68 amino acids lost. Probably it causes the enzyme activity seriously dropped down and being pathologically the basis of disease.

Of denaturing high performance liquid chromatography (DHPLC), a technique platform was developed for screening G6PD deficient variants.When applied to screen and identify the G6PD deficient variants from 124 patients who come from 11 nations in China, the DHPLC was compared with amplification refractory mutation system (ARMS) or DNA sequence technique and assessed carefully in its accuracy, sensitivity, efficiency and the cost of experiment.The G6PD-deficient variants, such as 1388 G--A (36/124 cases), 1376 G--T(35), 95 A--G (14), 1024 C--T (3), 392 G--T (4), 871 G--A /1311 C--T /IVS XI +93 t--c (9), 871 G--A (1), 1311 C--T/IVS XI +93 t--c (4), 1376 G--T /1388 G--A (1) and so on, were characterized as sharp peaks by DHPLC and verified by DNA sequence. Further, the standard chromatograms were put into database for 8 kinds of common G6PD deficient variants in Chinese populations. And also DHPLC found 3 G6PD variants (1388 G--A) from 103 negative controls. With taking 8.8 minutes and costing 1 dollar for each sample, DHPLC successfully detected and identified 34 heterozygous females from patients with G6PD deficiency. However, ARMS checked 83 positive controls but got 12 false G6PD mutants, of which 5 were false positive, 7 false negative. Above results show that DHPLC sounds like to be more convenience, sensitive and accurate than ARMS and DNA sequence techniques for checking G6PD mutants.DHPLC is of great advantage to screen the G6PD deficient variants with accuracy, convenience, automation and less cost, and significantly to identify the female heterozygote and clinical type IV individuals with G6PD deficiency.