MYO1E gene is located on chromosome 15 and encodes myosin 1e,which acts as an actin-based molecular motor of cytoskeleton. Myosin 1e is critical to maintain the podocyte function and the conse-quent integrity of the glomerular filtration barrier. Mutations in MYO1E gene has been indentified to be the cause of childhood-onset,familial steroid-resistant focal segmental glomerulosclerosis. Surprisingly,three patients with MYO1E mutations had partial remission by cyclosporine therapy. Detection of the MYO1E gene in the patients suffering from steroid-resistant nephrotic syndrome will be beneficial to make therapeutic decisions and predict prognoses.

Benign familial hematuria,also called thin basement membrane nephropathy,is caused by a heterozygous mutation in the COL4A3 or COL4A4 gene.The prognosis of the patients with benign familial hematuria,who present isolated hematuria without associated with proteinuria and normal renal function,is good in childhood.However,the prognosis of part of the patients with benign familial hematuria,who appear proteinuria,hypertension,chronic renal failure and end-stage kidney disease,is poor in adulthood.Therefore,the patients with benign familial hematuria should be carried on the long-term follow-up,and may be reviewed every 1-2 years for hypertension,proteinuria,and renal impairment.Treatment for benign familial hematuria should include an angiotensin converting enzyme inhibitor to delay the onset of renal failure.

Adolescent ; Child ; DNA Mutational Analysis ; Genetic Testing ; methods ; High-Throughput Nucleotide Sequencing ; Humans ; Infant ; Mutation ; Nephrotic Syndrome ; congenital ; diagnosis ; genetics ; Phenotype ; Polymerase Chain Reaction

Objective To examine mutations in the WT1 and PLCE1 gene in three Chinese families with autosomal recessive steroid-resistant nephrotic syndrome (SRNS) once mutations in NPHS2 had been excluded. Methods Peripheral blood samples were collected for genetic analysis from three probands of three Chinese families and their parents, and two probands' siblings, and 50 adult volunteers with normal urinalysis. Genomic DNA was isolated from peripheral blood leucocytes. Ten exons and exon-intron boundaries of WT1, and 31 exons and exon-intron boundaries of PLCE1 were amplified by polymerase chain reaction (PCR). Mutational analysis was performed by DNA sequencing directly and RFLP (restriction fragment length polymorphism) and/or PCR. Results No mutation in both WT1 and PLCE1 was identified in three probands from three Chinese families with autosomal recessive SRNS. However, three variants of WT1, 126C>T, ⅣS5-64A>G and 903A>G, and 13 variants of PLCE1, -134A>G, 810T>C, 960G>A, ⅣS11-28C>G, ⅣS15+26A>C, 4724G>C, ⅣS20+40C>T, ⅣS21+64G>A, ⅣS22-26T> A, 5320C>T, 5780A>G, ⅣS27+24A>G and ⅣS31 +48_49insT, were detected in three probands and some controls, indicating that all these variants were gene polymorphisms. WT1 polymorphism ⅣS5-64A>G, and PLCE1 polymorphism ⅣS22-26T>A were novel. Conclusion All the encoding exons and exon-intron boundaries of both WT1 and PLCE1 in three probands are examined, and no causative mutations in WT1 and PLCE1 axe found, suggesting that mutation in WT1 and PLCE1 genes is not a major cause of the Chinese families with autosomal recessive SRNS.

Objective To evaluate the efficacy of dezocine for prevention of postoperative cognitive dysfunction (POCD) in elderly patients undergoing total knee arthroplasty under remifentanil-based anesthesia.Methods Sixty-eight patients of both sexes, aged 65-85 yr, weighing 48-78 kg, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ , undergoing elective unilateral total knee arthroplasty under general anesthesia, were randomly divided into 2 groups (n =34 each) using a random number table: control group (group C) and dezocine group (group D).After induction of anesthesia, the patients were tracheally intubated and mechanically ventilated.Anesthesia was maintained with iv infusion of propofol 0.05-0.06 mg · kg-1 · min-1 and remifentanil 0.05-0.10 μg · kg-1 · min-1, and intermittent iv boluses of vecuronium 0.04 mg/kg.Dezocine 0.1 mg/kg was injected intravenously at 30 min before skin incision in group D.At 1 day before operation (T0) , and 1, 3, 5, 7 days after operation (T1 , T2, T3 , T4) , the blood samples from the central vein were collected for determination of serum cortisol (Cor), neuron specific enolase (NSE) and S-100β protein concentrations.Before operation, and 5 and 7 days after operation, the patients' cognitive function was assessed using Mini-Mental State Examination, and the occurrence of POCD was recorded.Results Compared with the baseline value at T0, the serum Cor, NSE and S-100β protein concentrations were significantly increased at T1-3, and MMSE scores were decreased at T3,4 in the two groups.Compared with group C, the serum Cot, NSE and S-100β protein concentrations were significantly decreased at T1-4, and Mini-Mental State Examination scores were increased at 5 and 7 days after operation, and the incidence of POCD was decreased in group D.Conclusion Dezocine 0.1 mg/kg intravenously injected at 30 min before skin incision can prevent the occurrence of POCD in elderly patients undergoing total knee arthroplasty under remifentanil-based anesthesia.

Objective To elucidate the mutations of NPHS1 gene in children with sporadic steroid-resistant nephrotic syndrome (SRNS) in Southern Chinese Han ethnic group.Methods Peripheral blood samples were collected for genetic analysis from 40 patients with sporadic SRNS and 50 healthy volunteers as control.Genomic DNA was isolated from peripheral blood leucocytes.Twenty-nine exons and exon-intron boundaries of the NPHS1 gene were amplified by polymerase chain reaction.Mutational analysis was performed by DNA sequencing directly.Results Seven variants,928G＞A(D310N),2677A＞G (T893A),2869G＞C (V957L),IVS8+30C＞T,IVS21+14G＞A,IVS25-23C＞T and *142T＞C,of NPHS1 gene were found in 6 of 40 children with sporadic SRNS,whereas they were not found in 50 healthy controls.2677A ＞G,IVS8 +30C ＞T,IVS21 +14G＞A,IVS25-23C ＞T and *142T＞C were novel.Moreover,thirteen already reported NPHS1 polymorphisms,294C＞T,349G＞A,IVS3+15C＞T,IVS3+61A＞G,803G＞A,IVS8+68A＞G,1339G ＞A,1802G ＞C,2223C ＞T,2289C ＞T,IVS24 +36C ＞T,3315G＞A and IVS27 +45C ＞T,were detected in some patients and controls. Conclusions NPHS1 mutations in 6 of 40 children with sporadic SRNS in Southern Chinese Han ethnic group (15％) are detected.NPHS1 mutations are existed in Southern Chinese children,so it is necessary to perform the mutation analysis of NPHS1 gene in those children patients.

Objective To study the feasibility of testing three disease-causing genes for Alport syndrome,COL4A3,COL4A4 and COL4AS,in diagnosing patients with sporadic Alport syndrome by using targeted capture and next generation sequencing.Methods The clinical data of a 9-year-old boy suspected with Alport syndrome were collected.Genomic DNA was extracted using standard procedures from the peripheral blood leukocytes of the patient and his parents,respectively.Targeted capture and next generation sequencing and Sanger sequencing were applied to analyze the mutations in the 3 disease-causing genes.Clinical data of cases reported already with autosomal recessive Alport syndrome caused by the mutations in the COL4A4 gene were summarized.Results The patient was presented with neither family history of hematuria nor chronic renal failure.Haematuria and proteinuria were found at the age of 1 year.The patient presented with episodes of macrohaematuria and gradually developed nephrotic-level proteinuria.At the age of 8 years 7 months,bilateral sensorineural hearing loss was diagnosed.So a probable diagnosis of Alport syndrome was postulated.A compound heterozygous pathogenic mutations of 3578-1G ＞ A and 3967 C ＞ T(Q1323X) were identified in the COL4A4 gene in the patient.The mutation of 3578-1G ＞ A was inherited from his father,and the mutation of Q1323X from his mother.The patient was decisively diagnosed with autosomal recessive Alport syndrome.Conclusions The test of COL4A3,COL4A4 and COL4A5 genes can help diagnose patients with sporadic Alport syndrome by using targeted capture and next generation sequencing.

Renal glycosuria (RG) is an inherited disorder due to defective reabsorption of glucose by the proximal renal tubular.It is attributed to the mutations in the SCL5A2 gene,encoding the sodium-glucose transporter 2 (SGLT2).A defect of SGLT2 is responsible for impaired reabsorption of the filtered glucose in the proximal renal tubular,termed S1,which leads to glycosuria.RG is characterized by normal fasting serum glucose concentration and persistent isolated glucosuria,identification of glucose as the urinary sugar.The inherited pattern of RG is co-dominant inheritance trait with incomplete penetrance.The diagnostic criteria of glycosurias are as follows:a constant and relatively stable urinary glycosurias (10-100 g/d),identification of glucose as the urinary sugar,normal concentration of fasting plasma glucose and normal oral glucose tolerance test,evidence that individuals have normal carbohydrates intake,storage and metabolism.RG does not require special treatment generally,but the advice concerns diet with increasing the intake of carbohydrates.Physical activity should be moderate and professional,and excesive muscle and exercise should be not advisable.

Objective To establish a high performance liquid chromatography method to detect the concentration of lamotrigine in blood dialysate and investigate in vitro recovery of lamotrigine and the factors.Select the microdialysis conditions that apply to the animal experiment and guide the stability study of in vivo recovery.Methods Positive dialysis and retrodialysis were used for the examination of lamotrigine in vitro recovery and the influencing factors such as flow rate, concentration, temperature and time.Filtered out the best conditions that apply to the in vivo experiment.Used the retrodialysis to determine the in vivo recovery and its stability.Results There was no significant difference between relative recovery and relative loss in the same flow rate.The concentration had no obvious effect on relative recovery.At the same condition,relative recovery decreased with the increase of the flow rate and increased with the temperature.The in vivo recovery had a good stability of 6.5 hours when the flow rate and stabilization time were set at 2μL/min and 1.5 h, respectivily.Conclusion Microdialysis technique can be used for the pharmacokinetic study of lamotrigine.Retrodialysis can be used for the determination of the lamotrigine in vivo recovery.

To study the relation between drug release and the drug status within curcumin-loaded microsphere, SPG (shirasu porous glass) membrane emulsification was used to prepare the curcumin-PLGA (polylactic-co-glycolic acid) microspheres with three levels of drug loading respectively, and the in vitro release was studied with high-performance liquid chromatography (HPLC). The morphology of microspheres was observed with scanning electron microscopy (SEM), and the drug status was studied with X-ray diffraction (XRD), differential scanning calorimetry (DSC) and infrared analysis (IR). The drug loading of microspheres was (5.85 ± 0.21)%, (11.71 ± 0.39)%, (15.41 ± 0.40)%, respectively. No chemical connection was found between curcumin and PLGA. According to the results of XRD, curcumin dispersed in PLGA as amorphous form within the microspheres of the lowest drug loading, while (2.12 ± 0.64)% and (5.66 ± 0.07)% curcumin crystals was detected in the other two kinds of microspheres, respectively, indicating that the drug status was different within three kinds of microspheres. In the data analysis, we found that PLGA had a limited capacity of dissolving curcumin. When the drug loading exceeded the limit, the excess curcumin would exist in the form of crystals in microspheres independently. Meanwhile, this factor contributes to the difference in drug release behavior of the three groups of microspheres.