Objective To study the lysozyme activity in Oncomelania hupensis and observe its inhibitory effect on bacterial growth.Methods Soft tissues of Oncomelania hupensis were initially homogenized and immersed in Tris-HCl-TritonX-114 buffer solution for 24 hours then the supernatant was collected after centrifugation at 10 000 × g for 10 minutes.The supernatant was incubated in a 37 ℃ water bath for 15 minutes and centrifuged again at 2000 × g for 10 minutes.The precipitate was put into ultrafiltration tube (relative retention molecular mass =3000) and centrifuged at 4 ℃,5 000 × g for 30 minute to obtain concentrated enzyme.The protein content,lysozyme activity and the antibacterial effect on Micrococcus lysodeikticus,Shigella dysenteriae,Staphylococcus aureus,Escherichia coli and Candida albicans were measured with bicinchonininc acid(BCA) method,turbidimetric method and agar diffusion (K-B) method,respectively.Results The antibacterial protein lysozyme was identified in gastropod protein concentration of the concentrated enzyme was 3.428 g/L.Average activity,total activity,and specific activity were (760 ± 120) × 103 U/L,(1520 ± 240) × 103 U/L and (221.70 ± 35.00)U/mg,respectively.The enzyme had produced exclusive inhibitory effects on growth of Micrococcus lysodeikticus and Shigella dysenteriae.Average inhibitory diameters were 10-12 and 12-15 mm,respectively.No inhibition zone was observed in saline control,Staphylococcus aureus,Escherichia coli and Candida albicans.Conclusions Lysozyme can be extracted from soft tissues of Oncomelania hupensis with Tris-HCl-TritonX-114 buffer solution,and the enzyme has inhibitory effect on growth of Micrococcus lysodeikticus and Shigella dysenteriae but has no antibacterial effect on Staphylococcus aureus,Escherichia coli and Candida albicans.

The infant (a girl aged 6 months) was admitted to the hospital because of oliguria and acute renal dysfunction. The laboratory examination results showed serious metabolic acidosis and increased blood urea nitrogen and serum creatinine levels. The patient continued to be anuric after 10 days of treatment with continuous renal replacement therapy (CRRT). she died a day later. The family history showed that the patient's sister died of acute renal failure 6 months after birth. The genomic sequencing results showed AGXT mutation in the patient and confirmed the diagnosis of primary hyperoxaluria type 1 (PH1). Her parents were heterozygous carriers. PH1 should be considered when the children have abnormal renal function or recurrent renal calculi or have a family history of these symptoms. AGXT gene analysis is an important method for PH1 diagnosis.
Acute Kidney Injury ; etiology ; Female ; Humans ; Hyperoxaluria, Primary ; complications ; Infant ; Mutation ; Oliguria ; etiology ; Transaminases ; genetics

From an aqueous extract of Lonicera japonica flower buds, sixteen compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, MCI gel, silica gel, and sephadex LH-20 and reversed-phase HPLC. Their structures were elucidated by spectroscopic data analysis as 6'-O-acetylvogeloside (1), 6'-O-acetylsecoxyloganin (2), dichlorogelignate (3), guanosinyl-(3' --> 5')-adenosine monophosphate(GpA,4) , 5'-O-methyladenosine (5), 2'-O-methyladenosine (6), adenosine (7), syringin (8), methyl 4-O-β-D-glucopyranosyl caffeate (9), (-)-dihydrophaseic acid 4'-O-β-D-glucopyranoside (10), ketologanin (11), 7α-morroniside (12), 7β-morroniside (13), kingiside (14), cryptochlorogenic acid methyl ester (15), and 6-hydroxymethyl-3-pyridinol (16). All the compounds were obtained from this plant for the first time, compounds 1 and 2 are new compounds, 3 and 5 are new natural products, and 4 is the first example of dinucleoside monophosphate isolated from a plant extract.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flowers ; chemistry ; Lonicera ; chemistry ; Mass Spectrometry ; Molecular Structure

A boy aged 11 years was admitted due to intermittent weakness and difficulty in walking for 6 years, and hepatomegaly, glycopenia and unconsciousness for 4 years. The laboratory examinations showed severe metabolic acidosis, hypoglycemia, and abnormal liver function. CT scan showed marked liver enlargement with fat density shadow. The boy was given fluid infusion, correction of acidosis, intravenous injection of glucose, L-carnitine, compound vitamin B, and coenzyme Q10, but he was in a persistent coma and it was difficult to correct refractory metabolic acidosis and hypoglycemia. The boy died. Blood and urinary organic acid screening and gene detection confirmed that the boy had late-onset glutaric aciduria type II (GAIIc) caused by electron-transferring-flavoprotein dehydrogenase (ETFDH) gene defect. GAIIc is an inherited metabolic disease with a low incidence, resulting in a high misdiagnosis rate. GAIIc should be considered for children with recurrent weakness or reduced activity endurance, hypoglycemia, and marked liver enlargement with abnormal liver function. Urinary organic acid analysis and blood tandem mass spectrometry can help with the early diagnosis of GAIIc, and ETFDH gene analysis helps to make a confirmed diagnosis.
Child ; Hepatomegaly ; etiology ; Humans ; Hypoglycemia ; etiology ; Male ; Multiple Acyl Coenzyme A Dehydrogenase Deficiency ; diagnosis ; Muscle Weakness ; etiology

Objective To investigate the methods and clinical significance of breast cancer treated with photodynamic.Methods From June to December in 2005,photodynamic therapy was used in 12 cases confirmed intramammary lymph node metastasis before operation and 15 cases confirmed chest wall recur- rences by means of lymph node imaging.Results The intramammary lymph node metastasis whose diameter between 0.5～1.0cm measured by lymph node imaging preoperatively completely disappeared when rechecked 3 months postoperatively.Chest wall recurrence regions of breast cancer whose diameter less than 1.0 cm completely remitted.Conclusion Photodynamic therapy is helpful to eliminate the intramammary lymph node metastasis and to cure the postoperative chest wall recurrence of breast cancer.

OBJECTIVETo explore the correlations of dopamine transporter gene (DAT) and dopamine D(2) receptor gene (DRD2) to stuttering.

METHODSTo examine the correlations of the 5 single nucleotide polymorphisms (SNPs) in dopaminergic gene (C252T, C1804T, and C1820T in DAT gene, and T1054C and C1072T in DRD(2) gene) to stuttering in Han Chinese individuals, a case-control study involving 112 patients with stuttering and 112 gender-matched controls was carried out. Genotyping was performed by a combined approach using polymerase chain reaction (PCR) and pyrosequencing.

RESULTSC1804T showed no polymorphism in either the patients or the control subjects and was therefore excluded from the following analysis. The C allele frequency at C1072T site was significantly higher, but T allele frequency significantly lower in the stuttering group than in the control group. The patients had significantly higher CC and lower CT genotype frequencies than the control group. There were no significant differences in the allelic frequencies of C252T, C1820T and T1054C between the patients and the controls, suggesting a Hardy-Weinberg equilibrium at these 3 loci.

CONCLUSIONThe presence of the C allele at C1072T in DRD(2) gene is associated with increased susceptibility to stuttering in Han Chinese, whereas the T allele provides protection against the onset of stuttering.

Adult ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; China ; ethnology ; Dopamine Plasma Membrane Transport Proteins ; genetics ; Female ; Genotype ; Humans ; Male ; Polymorphism, Single Nucleotide ; Receptors, Dopamine D2 ; genetics ; Stuttering ; genetics ; Young Adult

OBJECTIVETo analyze the relationship between estrogen dependence of breast cancer cells and expression level of PC-cell derived growth factor (PCDGF) and investigate the possibility of practicing endocrine therapy in estrogen receptor negative breast cancer.

METHODSExpression level of PCDGF mRNA was detected by fluorescence quantitative polymerase chain reaction and cell growth curve was drawn by cell count kit-8 method, then analyzed the relationship between the expression of PCDGF and dependence of estrogen. PCDGF shRNA vector was constructed and transfected into breast cancer cell lines (MCF-7 and MDA-MB-231) using Lipofectamin 2000, examined and compared the changes of estrogen dependence between the two cell lines.

RESULTSPCDGF was expressed in most breast cancer cell lines, and the growth dependence of estrogen was higher in the cells with high-expressing PCDGF than those with low-expressing PCDGF. Expression of PCDGF could be inhibited by transfected shRNA into MCF-7 and MDA-MB-231 (inhibition rates were 81.1% and 86.7% respectively), inhibition of the expression of PCDGF could lead to higher growth dependence of estrogen, and the dependence of MDA-MB-231 was increased much more than MCF-7(P < 0.05).

CONCLUSIONSThe expression level of PCDGF is higher in estrogen receptor negative breast cancer than in estrogen receptor positive breast cancer. Inhibiting the expression of PCDGF could reverse resistance of endocrine therapy especially to estrogen receptor negative breast cancer.

Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; Estrogens ; pharmacology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptors, Estrogen ; metabolism ; Transfection

OBJECTIVETo investigate the myocardial protective effect of L-carnitine in children with hand, foot and mouth disease (HFMD) caused by Coxsackie A16 virus and possible mechanisms.

METHODSA total of 60 HFMD children with abnormal myocardial enzyme after Coxsackie A16 virus infection were enrolled and randomly divided into L-carnitine group and fructose-1,6-diphosphate group (fructose group), with 30 children in each group. The two groups were given L-carnitine or fructose diphosphate in addition to antiviral and heat clearance treatment. Another 30 healthy children who underwent physical examination were enrolled as control group. The changes in myocardial zymogram, malondialdehyde (MDA), superoxide dismutase (SOD), and apoptosis factors sFas and sFasL after treatment were compared between groups.

RESULTSThere was no significant difference in treatment response between the L-carnitine group and the fructose group (P>0.05). One child in the fructose group progressed to critical HFMD, which was not observed in the L-carnitine group. Before treatment, the L-carnitine group and the fructose group had significantly higher indices of myocardial zymogram and levels of MDA, sFas, and sFasL and a significantly lower level of SOD than the control group (P<0.05), while there were no significant differences in these indices between the L-carnitine group and the fructose group (P>0.05). After treatment, the L-carnitine group and the fructose group had significant reductions in the indices of myocardial zymogram and levels of MDA, sFas, and sFasL and a significant increase in the level of SOD (P<0.05); the fructose group had a significantly higher level of creatine kinase (CK) than the control group and the L-carnitine group, and there were no significant differences in other myocardial enzyme indices, MDA, sFas, and sFasL between the L-carnitine group and the fructose group, as well as between the L-carnitine and fructose groups and the control group (P>0.05). SOD level was negatively correlated with aspartate aminotransferase, lactate dehydrogenase (LDH), CK, and creatine kinase-MB (CK-MB) (r=-0.437, -0.364, -0.397, and -0.519 respectively; P<0.05), and MDA level was positively correlated with LDH and CK-MB (r=0.382 and 0.411 respectively; P<0.05).

CONCLUSIONSL-carnitine exerts a good myocardial protective effect in children with HFMD caused by Coxsackie A16 virus, possibly by clearing oxygen radicals and inhibiting cardiomyocyte apoptosis.

Carnitine ; therapeutic use ; Child, Preschool ; Coxsackievirus Infections ; complications ; Female ; Hand, Foot and Mouth Disease ; drug therapy ; etiology ; metabolism ; Heart ; drug effects ; Humans ; Infant ; Male ; Malondialdehyde ; analysis ; Myocardium ; metabolism ; pathology ; Superoxide Dismutase ; metabolism

Objective To investigate the effects of advanced glycation end-products(AGEPs)on the function of normal keratinocytes in vitro so as to explore the role of AGEPs in impaired wound healing. Methods Normal rat keratinocytes were incubated with different concentrations of AGEPs.After 48 hours of culturing,the cell proliferation rates were measured by MTT colorimetric determination.The cell cycle distributions and apoptosis were analyzed with flow cytometry,and the migration was investigated by 24-well fluorimetric cell migration assay kit by exposing to 100?g/ml AGEPs.Nuclear extracts from these cells were examined for binding of nucleotides containing NF-?B consensus by immunocytochemistry and EMSA in vitro.Results The proliferations of normal keratinocytes were significantly arrested and many cells were induced to early apoptosis compared with control ones(P＜0.05)by exposing to AGEPs for 48 hours. Meanwhile AGEPs also irritated keratinocytes migration compared with control ones(P＜0.05).Inhibiting the activation of NF-?B could partly recover the proliferation of keratinocytes,reverse apoptosis and attenu- ate migration.Conclusion AGEPs are correlated with the migration,proliferation and apoptosis of kera- tinocytes by NF-?B.

Objective To observe the clinical efficacy of herb-partitioned fire therapy in treating stomachache due to cold-deficiency. Method Sixty patients with stomachache due to cold-deficiency were randomized into a treatment group and a control group, 30 cases each. The control group was intervened by Western and Chinese medications, based on which, the treatment group was given herb-partitioned fire therapy. The clinical efficacies of the two groups were observed, as well as the symptoms scores before and after the treatment. Result There was a significant difference in the clinical efficacy between the two groups (P<0.01). The symptoms scores dropped after the intervention in both groups, and the score in the treatment group was significantly lower than that in the control group (P<0.01). Conclusion Herb-partitioned fire therapy plus medication is effective in treating stomachache due to cold-deficiency, and it can produce a more significant efficacy than medication alone.