Adolescent ; Child ; Diagnosis, Differential ; Dystonic Disorders ; diagnosis ; drug therapy ; physiopathology ; Female ; Humans ; Male

OBJECTIVEMutations 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.

METHODSThe 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.

RESULTSThe 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.

CONCLUSIONA 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.

Amino Acid Sequence ; Asian Continental Ancestry Group ; Base Sequence ; Child ; China ; Chromatography, High Pressure Liquid ; Genotype ; Humans ; Iduronate Sulfatase ; genetics ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis II ; diagnosis ; genetics ; Mutation ; Reverse Transcriptase Polymerase Chain Reaction

Internal lenticonus is a very rare morphologic abnormality of crystalline lens which has been reported in only several cases in the literature. We herein reported the clinical characteristics and surgical findings of the anterior internal lenticonus accompanied by congenital nuclear cataract. Cataract extraction accompanied with intraocular lens implantation was uneventfully performed, and a good visual outcome was achieved in this case. Viral infection during embryonal and fetal period might account for the formation of the anterior internal lenticonus and congenital nuclear cataract in our case.
Cataract Extraction ; methods ; Humans ; Lens Diseases ; surgery ; Lens Implantation, Intraocular ; methods ; Lens, Crystalline ; abnormalities ; surgery ; Male ; Middle Aged ; Visual Acuity

OBJECTIVETo observe the effect and safety of plastering Chinese Compound Shenhuang Ointment (CSO) at Shenque (RN8) in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of qi stagnation blood stasis syndrome (QSBSS).

METHODSA prospective, multi-centered, randomized, double-blinded, controlled trial was conducted in 220 postoperative gastrointestinal dysfunction patients of QSBSS. They were randomly assigned to two groups, the CSO group (110 cases) and the placebo group (110 cases). CSO was plastered at Shenque (RN8) for 5 days after operation. The time of exhaustion, defecation, the recovery of intestinal peristalsis, integrals of TCM syndrome, and serum levels of motilin (MOT)and somatostatin (SS) were observed.

RESULTSCompared with the placebo group, the condition of exhaustion and defecation, the recovery of intestinal peristalsis on the 3rd day after operation was all improved (P < 0.05). The integrals of TCM syndrome at day 2, 3, and 4 were more significantly lowered in the CSO group than in the placebo group (P < 0.01, P < 0.05). The total effective rate of TCM syndrome was 95.3% in the CSO group, better than that in the placebo group (91.8%, P < 0.05). Compared with the placebo group, the serum MOT level increased and the serum SS level decreased at day 5 after operation in the CSO group (P < 0.05).

CONCLUSIONSThe plastering of CSO at Shenque (RN8) could advance the time of exhaustion and defecation, and improve patients' clinical symptoms. And patients could tolerate well.

Aged ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gastrointestinal Diseases ; drug therapy ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Ointments ; Postoperative Period ; Prospective Studies

Objective:To compare the differences of anatomical and histological characteristics of lymph nodes between BALB/c nude mice and BALB/c mice.Methods:Firstly,twenty BALB/c nude mice and twenty BALB/c mice were dissected by using a surgical microscope.Secondly,the differences of T cells and B cells at the lymph node were compared by the expressions of CD3 and CD20 immunohistochemistry dyes.Results:There were,on average,23 nodes per mouse contained within the large lymph node assembly in the BALB/c nude mouse.The anatomical features of the lymph node distribution in the nude mice were mainly found in the neck with relatively higher density.There were two lymph nodes both in the submandible lymph nodes group and in the superficial cervical lymph nodes group (the constituent ratios were 95％ and 90％,respectively) in the BALB/c nude mice,but there were four lymph nodes (the constituent ratios were 95％ and 90％,respectively) in the BALB/c mice.There were significant difference between the BALB/c nude mice and the BALB/c mice.Mostly there were two lymph nodes of deep cervical lymph nodes both in the BALB/c nude mice and the BALB/c mice (the constituent ratios were 95％ and 100％,respectively).There were no significant difference between the BALB/c nude mice and the BALB/c mice.We confirmed that the number of CD3-positive T lymphocytes in lymph nodes of the nude mice decreased greatly as compared with the BALB/c mice.Expressions of CD3 in T cells were 95％ and 100％ in the BALB/c nude mice and in the BALB/c mice,respectively.There were significant differences between the BALB/c nude mice and the BALB/c mice.Expressions of CD2o in B cells were 95％ and 100％ in the BALB/c nude mice and in the BALB/c mice,respectively.There was no significant difference between the BALB/c nude mice and BALB/c mice.Conclusion:The anatomi cal pictures of lymph node distribution in the nude mouse will be benefit to those who are interested.The anatomical features of the lymph node local higher density in neck of the nude mouse and lack of CD3-positive T lymphocytes would be useful for obtaining a better understanding of localized lymph node metastasis of oral transplant tumors.

OBJECTIVETo identify the mutations of iduronate-2-sulfatase (IDS) gene, and to establish a basis of prenatal gene diagnosis of Hunter syndrome.

METHODSUrine 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.

RESULTSAbnormal 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.

CONCLUSIONA 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.

Child, Preschool ; Chromatography, High Pressure Liquid ; DNA Mutational Analysis ; Humans ; Iduronate Sulfatase ; genetics ; Male ; Molecular Sequence Data ; Mucopolysaccharidoses ; genetics ; Mucopolysaccharidosis II ; enzymology ; genetics ; Mutation

In vitro percutaneous delivery of hepatitis B vaccines was investigated in order to assess the penetration of vaccine under passive diffusion and iontophoresis conditions. The study was carried out using Franz vertical diffusion cell through the hairless abdominal skin of Sprague-Dawley (SD) rats. Enzyme-linked immunosorbent assay (ELISA) was used to determine the cumulative amount of permeation and the retention amount of drug in skin. Passive diffusion alone resulted in less skin permeation and retention of hepatitis B vaccines, only (2.1 +/- 0.1) ng x cm(-2) and (2.3 +/- 0.1) ng x cm(-2) after 24 h when the initial concentration of vaccine in the donor compartment was 23 microg x mL(-1) and 46 microg x mL(-1), respectively. After removing the stratum corneum, the permeation and retention amount of hepatitis B vaccines increased to (383.7 +/- 86.2) ng x cm(-2) and (16.8 +/- 4.6) ng x cm(-2), respectively, 171.6-folds and 2.1-folds more than that from its intact skin with the drug loaded at 46 microg x mL(-1). Iontophoresis induced a significant increase of cumulative and retention amount of hepatitis B vaccines through the skin (P < 0.05). Application of iontophoresis significantly enhanced the permeation of hepatitis B vaccines (P < 0.05) by 2.7-folds and 6.6-folds for the intact skin, and by 1.6-folds and 1.8-folds for the tape-stripped skin with initial drug loading of 23 microg x mL(-1) and 46 microg x mL(-1), respectively. Iontophoresis also significantly increased the amount of drug retained in the skin. After applying iontophoresis for 6 h, the amount of skin retention was nearly the same as passive diffusion for 24 h both from intact skin [(16.8 +/- 4.6) ng x cm(-2) vs (13.3 +/- 5.4) ng x cm(-2)] (P > 0.05) and tape-stripped skin [(36.7 +/- 14.1) ng x cm(-2) vs (26.8 +/- 11.2) ng x cm(-2)] (P > 0.05). Overall, these findings revealed that the transportation efficiency of bioactive substance like hepatitis B vaccines may be improved by iontophoresis, which can be potentially used in the field of transcutaneous immunization.
Administration, Cutaneous ; Animals ; Diffusion ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B Vaccines ; administration & dosage ; pharmacokinetics ; In Vitro Techniques ; Iontophoresis ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Skin ; metabolism ; Skin Absorption

BACKGROUNDOral squamous cell carcinoma (OSCC) is the most prevalent malignant tumor in the head and neck region, comprising more than 90% of all oral malignancies. A feasible approach for an animal model to study OSCC lymph node metastasis was established and biological behaviors of three oral squamous cell carcinoma cell lines were compared.

METHODSAfter implanting three kinds of cell lines (GDC185, Tca8113, Tca83) into three different anatomical sites in nude mice, namely the tongue, floor of the mouth, and axillary fossa, we observed the tumorigenicity and the metastatic capacity, which was confirmed by histopathology under a surgical microscope.

RESULTSThe animal model injected with GDC185 cells into the floor of the mouth had the highest rate of neck lymph node metastasis (55.6%) and the cell lines had significantly different biological behaviors.

CONCLUSIONSNude mice injected with GDC185 cells into the floor of the mouth could be used as a feasible animal model to study neck metastasis of oral squamous cell carcinoma.

Animals ; Carcinoma, Squamous Cell ; pathology ; Cell Line, Tumor ; Humans ; Lymphatic Metastasis ; Mice ; Mice, Inbred BALB C ; Mouth Neoplasms ; pathology ; Neck

OBJECTIVETo investigate the association of single nucleotide polymorphisms (SNPs) of 1772 (C-->T) and 1790 (G-->A) in exon 12 of hypoxia-inducible factor 1, alpha subunit gene (HIF1A) with hypoxia adaptation in high altitude in Sherpas.

METHODSThe blood samples were chosen from 148 Sherpas in Tibet high altitude and 90 Han nationality healthy people in Guangdong province, and from which genomic DNA was extracted. The single nucleotide polymorphisms of 1772(C-->T) and 1790(G-->A) in exon 12 of HIF1A gene were examined by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR).

RESULTSThe genotype frequency of HIF1A gene 1790 (G-->A) in Sherpas and Han nationality was 57.43% versus 75.56% in GG genotype, 37.84% versus 21.11% in GA genotype and 4.73% versus 3.33% in AA genotype. GG genotype frequency in Sherpas was lower than that in Han nationality (P<0.01), while GA genotype frequency in Sherpas was higher than that in Han nationality (P<0.01). No significant difference in CC, CT and TT genotype frequency of 1772(C-->T) was shown between two groups respectively. The total frequency of CC + GA, CT + AA, TT + GA and TT + AA in Sherpas was higher than that in Han nationality.

CONCLUSIONPolymorphisms of HIF1A gene 1790 (G-->A) are associated with hypoxia adaptation in high altitude in Sherpas. GA and AA genotype may be benefit to hypoxia adaptation, and it is worthy of deep-going investigation.

Adaptation, Physiological ; genetics ; physiology ; Altitude ; Gene Frequency ; Genotype ; Humans ; Hypoxia ; physiopathology ; Hypoxia-Inducible Factor 1 ; genetics ; Linkage Disequilibrium ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; genetics ; Tibet

The hydroxycamptothecin (HCPT) PEGylated liposomes (HCPT-LP) were modified with RGD cyclopeptide formed the tumor-targeting liposomes (HCPT-RGD-LP). HCPT-LP and HCPT-RGD-LP were injected intravenously with single dose of 5 mg x kg(-1) to rats. The drug concentration in plasma was determined and the pharmacokinetic behaviour was compared. The HCPT distribution in heart, liver, spleen, lung, kidney and plasma of mice was investigated following intravenous administration of HCPT-LP and HCPT injection. The nude mice implanted human hepatoma HepG2 cells were studied by in vivo imaging. The fluorescent probe was DiR and the nude mice were injected with DiR PEGylated liposomes (DiR-LP) and DiR-LP modified with RGD cyclopeptide (DiR-RGD-LP). The results showed that there was no significant difference (P > 0.05) of main pharmacokinetic parameters t1/2beta, CL, V(c), AUC(0-48 h), AUC(0-inifinity), MRT(0-48 h), MRT(0-infinity) between HCPT-RGD-LP and HCPT-LP. HCPT-LP had a remarkably better long-circulating effect than HCPT injection in mice and the concentration of HCPT was highest in liver. The DiR accumulation in tumors of DiR-RGD-LP was higher than that of DiR-LP by the visualized fluorescence of in vivo imaging. It indicated that such PEGylated liposomes modified with RGD cyclopeptide could improve the tumor targeting efficacy.
Animals ; Area Under Curve ; Camptothecin ; administration & dosage ; analogs & derivatives ; chemistry ; pharmacokinetics ; Diagnostic Imaging ; Drug Delivery Systems ; Female ; Fluorescent Dyes ; Hep G2 Cells ; Humans ; Liposomes ; administration & dosage ; chemistry ; pharmacokinetics ; Liver Neoplasms ; diagnosis ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Oligopeptides ; administration & dosage ; chemistry ; pharmacokinetics ; Polyethylene Glycols ; administration & dosage ; chemistry ; pharmacokinetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectroscopy, Near-Infrared ; Tissue Distribution