Using Liquid Chromatography-Tandem Mass Spectrometry in Detecting Plasma Lyso-GL3 Levels in Patients with Fabry Disease and the Association Analysis of Phenotype-Genotype of the Disease / 罕见病研究

ObjectiveUsing the liquid chromatography-tandem mass spectrometry (LC-MS/MS) to determine the plasma level of Lyso-GL3 in patients with Fabry disease and to analyze the clinical application of the method.MethodsThirty-nine patients with a genetic diagnosis of Fabry disease were included, and plasma levels of Lyso-GL3 were measured by LC-MS/MS analysis, and detailed clinical information of the patients was obtained including: α-galactosidase A activity, genetic variants, quantification of urine protein, mean arterial pressure, and estimation of glomerular filtration rate, and the differences in the levels of Lyso-GL3 in different clinical phenotypes and genotypes were statistically analyzed, as well as the association with clinical indicators.ResultsLyso-GL3 showed good linearity within 0.7856-400 ng/mL(r=0.9992).Further analysis of 39 Fabry disease patients diagnosed in Ruijin Hospital, Shanghai Jiao Tong University School of Medicine showed a median Lyso-GL3 concentration of 23.6 ng/mL(4.3-92.9 ng/mL); Lyso-GL3 levels were significantly higher in patients with both the frameshift and the splicing mutations, as well as in patients with the nonsense mutations, than in patients with the missense mutations (median value 119.7 ng/mL vs. 11.9 ng/mL, P=0.006, and median value 97.0 ng/mL vs. 11.9 ng/mL, P=0.015, respectively). Whereas, association analysis revealed that Lyso-GL3 was not significantly associated with urinary protein, mean arterial pressure and estimated glomerular filtration rate.ConclusionsThe using of LC-MS/MS to quantify plasma Lyso-GL showed significant differences in Lyso-GL3 concentrations between classical and atypical phenotypes, suggesting that plasma Lyso-GL3 may help with clinical phenotypes. However, Lyso-GL3 levels is found to be overlapped between genotypes. No significant linear correlation was found between Lyso-GL3 and renal clinical indicators, suggesting the urgent need in finding a more accurate tool to assess renal involvement and prognosis in patients with Fabry disease.

Clinical value of ranibizumab combined with fundus laser in the treatment of macular edema secondary to branch retinal vein occlusion / 中国基层医药

Objective@#To investigate the clinical value of ranibizumab combined with fundus laser in the treatment of macular edema secondary to branch retinal vein occlusion(RVO).@*Methods@#From June 2016 to June 2017, 98 patients with RVO secondary macular edema in the People's Hospital of Linfen were randomly divided into three groups according to the digital table: A group (30 cases) treated with simple fundus laser, B group (33 cases) treated with intravitreal injection of ranibizumab, C group(35 cases) treated with fundus laser combined with intravitreal injection of ranibizumab.The best corrected visual acuity(BCVA), macular fovea thickness(CMT), leakage rate, injection times and complications were compared among the three groups before and after treatment.@*Results@#At 3, 6, 9, 12 months after treatment, the BCVA among A group, B group and C group had statistically significant differences (F=4.165, 5.021, 6.954, all P<0.05), and the BCVA of C group was better than that of B group, the BCVA of B group was better than that of A group (t=3.985, 3.852, 3.779, 4.021, 3.624, 3.729, all P<0.05). There were statistically significant differences in CMT among A group, B group and C group at 3 months and 6 months after treatment (F=6.772, 14.025, all P<0.05), and the CMT of C group was less than that of B group, the CMT of B group was less than that of A group (t=5.325, 11.251, 3.992, 6.895, all P<0.05). At 6 months after treatment, the leakage rate in C group (2.86%) was lower than that in B group (18.18%), the leakage rate in B group was lower than that in A group (23.33%) (χ2=6.148, P<0.05). The injection times of ranibizumab in B group was (2.93±1.52), which was significantly less than that in C group (2.00±0.56)(t=3.349, P<0.05).@*Conclusion@#The short-term clinical effect of ranibizumab combined with fundus laser in the treatment of macular edema secondary to RVO is better than laser therapy and ranibizumab alone, and the injection times of ranibizumab can be reduced.

Clinical value of ranibizumab combined with fundus laser in the treatment of macular edema secondary to branch retinal vein occlusion / 中国基层医药

Objective To investigate the clinical value of ranibizumab combined with fundus laser in the treatment of macular edema secondary to branch retinal vein occlusion (RVO).Methods From June 2016 to June 2017,98 patients with RVO secondary macular edema in the People's Hospital of Linfen were randomly divided into three groups according to the digital table:A group (30 cases) treated with simple fundus laser,B group (33 cases) treated with intravitreal injection of ranibizumab,C group(35 cases) treated with fundus laser combined with intravitreal injection of ranibizumab.The best corrected visual acuity (BCVA),macular fovea thickness (CMT),leakage rate,injection times and complications were compared among the three groups before and after treatment.Results At 3,6,9,12 months after treatment,the BCVA among A group,B group and C group had statistically significant differences (F=4.165,5.021,6.954,all P ＜0.05),and the BCVA of C group was better than that of B group,the BCVA of B group was better than that of A group (t =3.985,3.852,3.779,4.021,3.624,3.729,all P ＜0.05).There were statistically significant differences in CMT among A group,B group and C group at 3 months and 6 months after treatment (F =6.772,14.025,all P ＜ 0.05),and the CMT of C group was less than that of B group,the CMT of B group was less than that of A group (t =5.325,11.251,3.992,6.895,all P ＜ 0.05).At 6 months after treatment,the leakage rate in C group (2.86％) was lower than that in B group (18.18％),the leakage rate in B group was lower than that in A group (23.33％) (x2 =6.148,P ＜ 0.05).The injection times of ranibizumab in B group was (2.93 ± 1.52),which was significantly less than that in C group (2.00 ± 0.56) (t =3.349,P ＜ 0.05).Conclusion The short-term clinical effect of ranibizumab combined with fundus laser in the treatment of macular edema secondary to RVO is better than laser therapy and ranibizumab alone,and the injection times of ranibizumab can be reduced.

Observation on clinical effects of Zhu Lian's type Ⅰ excitation needling technique for postpartum urinary retention / 针灸推拿医学（英文版）

Objective: To observe the clinical effects of Zhu Lian's type Ⅰ excitation needling technique for postpartum urinary retention. Methods: A total of 60 cases with postpartum urinary retention were recruited and divided randomly into an observation group and a control group, 30 cases in each group. The control group was treated with routine acupuncture, and stimulated with sparse and dense wave of electric acupuncture for 15 min after arrival of needling sensation, and then the needles were taken out. The observation group was treated with Zhu Lian's type Ⅰ excitation needling technique, by inserting the needles with the quick inserting method, swift and temporary lifting and thrusting technique for shallow insertion for 5 times, by an in-and-out technique, without retaining the needles. Results: The total effective rate was 96.7% in the observation group and 83.3% in the control group. The difference in the total effective rate between the two groups was statistically significant (P<0.05). After the treatment, the first urination time was shorter in the observation group than that in the control group, with a statistical significance (P<0.01). After the treatment, the volume of residual urine after the first urination was less in the observation group than that in the control group (P<0.01). Conclusion: Zhu Lian's type Ⅰ excitation needling technique can effectively promote the voluntary urination and bladder emptying in patients with postpartum urinary retention, and it takes effect faster.

Clinical Study on ZHU Lian's Acupuncture Activating Method for Hypotonic Cerebral Palsy / 上海针灸杂志

Objective To observe the clinical efficacy of ZHU Lian's acupuncture activating method with type 1 manipulations in treating hypotonic cerebral palsy.Method Sixty patients with hypotonic cerebral palsy, aged 9-59 months, were randomized into a treatment group and a control group, 30 cases in each group. The treatment group was intervened by ZHU Lian's acupuncture activating method with type 1 manipulations, while the control group was intervened by acupoint injection of vitamin B1 and B12. The two groups were treated once a day, 5 sessions as a treatment course. The Gross Motor Function Measure-88 (GMFM-88) was evaluated before the treatment and after 10 treatment courses.Result The GMFM-88 score was significantly changed after the treatment in both groups (P<0.05). After the intervention, the GMFM-88 score in the treatment group was significantly different from that in the control group (P<0.05).Conclusion ZHU Lian's acupuncture activating method is an effective approach in treating hypotonic cerebral palsy.

The clinicopathological characteristics of diabetic nephropathy and non-diabetic renal diseases in diabetic patients / 中华内科杂志

Objective To analyze the clinicopathological characteristics of renal lesions in type 2 diabetic patients and to differentiate diabetic nephropathy (DN) from non-diabetic renal diseases (NDRD).Methods Type 2 diabetic patients who received renal biopsy in Ruijin Hospital from January 2011 to December 2015 were recruited in this study. Clinical history, laboratory results and pathological data were retrospectively collected. According to the pathological findings, the patients were divided into 3 groups: DN, NDRD, DN+NDRD. Logistic model was applied to explore the independent clinical predictive factors in differentiating DN from NDRD. Results A total of 207 type 2 diabetic patients received renal biopsy, accounting for 6.82% of all biopsy population. Fifty-one patients were diagnosed with DN, 142 with NDRD and 14 with both DN and NDRD. In NDRD, membranous nephropathy(MN) (34.5%) was the most common finding, followed by IgA nephropathy(19.7%).By contrast, NDRD patients manifested a shorter diabetic course, a higher baseline hemoglobin level, a lower baseline serum creatinine, a higher prevalence of hematuria, a lower prevalence of hypertension and diabetic retinopathy, a better control of blood glucose, better compliance of monitoring blood glucose and less family history of diabetes. In multivariate logistic model, diabetic family history(OR=4.68, P=0.04) and long history of diabetes(OR=1.01, P=0.02) were risk factors of DN. Conclusion There is a high prevalence of NDRD in diabetic patients with renal lesions. Family history of diabetes and duration of diabetes are independent predictors of DN.

Influence ofZhulian acupuncture exciting method on hypoxic ischemic brain damage in rats to oxidative stress / 国际中医中药杂志

Objective To observate theZhulian acupuncture exciting method in different time of hypoxia ischemia brain injury in rat brain tissue malondialdehyde (MDA), monoamine oxidase (MAO), nitric oxide (NO) and glutathione peroxidase (GSH-PX) content.Methods 7 days old rats were randomly divided into a excitation method acupuncture group I, a excitation method acupuncture group II, a model group, a sham operation group, a normal control group, 10 rats in each group. Excitation method acupuncture group I and normal control group were given stimulation ofZhulian acupuncture exciting method one type technique from 24 h model manipulation, excitation method acupuncture II group from the beginning of the eighth day given Stimulation ofZhulian acupuncture exciting method one type technique. The sham operation group and the model group were not treated by acupuncture. The animals were sacrificed at the twenty-first day after making the model, determined brain tissue MDA、MAO、NO and GSH-PX Content.Results Compared with the model group, MDA (3.4 ± 0.87 nmol/mgvs. 5.50 ± 1.58 nmol/mg) content decreased in the excitation method acupuncture group I (P<0.05). The NO (12.43 ± 3.47μmol/mgvs. 17.10 ± 5.82μmol/mg) content decreased in the excitation method acupuncture group II (P<0.05). MAO (32.12 ± 11.15 U/mg, 31.01 ± 9.92 U/mgvs. 40.90 ± 11.02 U/mg) content were decreased in both excitation method acupuncture group I and group II (P<0.05), while the GSH-PX (2.61 ± 1.20 U/mg, 2.61 ± 1.37 U/mgvs. 1.43 ± 0.49 U/mg) content were increased (P<0.01). ConclusionZhulian acupuncture exciting method one type technique can decrease the content of MDA, MAO and NO reduce the content of hypoxic-ischemic brain injury in rat brain tissue, increase the content of GSH-PX, promote the removal of immature rats with hypoxic ischemic brain damage brain tissue metabolism, and protect brain function.

Effects of Doxofyline on Intraoperative Pulmonary Function in Patients Undergoing Double Lumen Endotracheal Intubation for One-lung Ventilation / 医药导报

Objective To evaluate the effects of doxofyline on intraoperative pulmonary function in patients receiving double lumen endotracheal intubation for one-lung ventilation. Methods Fifty patients who underwent elective pulmonary lo-bectomy under general anesthesia using double lumen endotracheal intubation were randomly divided into two groups ( n=25 each):control group (group C) and doxofyline group (group D).Doxofyline (4 mg?kg-1) was injected intravenously after double lumen endotracheal intubation in group D,while equal volume of 0.9% sodium chloride was intravenously given in group C.Total intravenous anesthesia with target controlled infusion was performed during the operation.Two milliliter blood samples were taken from the radial artery for blood gas analysis immediately before administration ( t0 ) ,at 30 min ( t1 ) ,60 min ( t2 ) after one-lung ventilation and at the moment of two-lung ventilation after chest closing ( t3 ) . The PaCO2 , PaO2 , peak airway pressure (Ppeak),airway plateau pressure (Pplat),airway resistance (Raw) and lung compliance (Compl) were recorded at t0-3. Results The Ppeak,Pplat and Raw were significantly decreased and the Compl and PaO2 significantly increased at t1-t3 in group D when com-pared with those in group C (P<0.05).The Ppeak,Pplat and Raw were significantly increased and Compl and PaO2 significantly de-creased at t3 as compared with those at t0 in group C ( P<0.05) . Conclusion Doxofyline can improve intraoperative pulmonary function in patients who undergo double lumen endotracheal intubation for one-lung ventilation.

Effect of Zhu Lian Acupuncture Exciting Method on Nerve Cell Apoptosis and the Expressions of PI3K, AKt and Caspase-3 Proteins in Young Rats with Hypoxic-ischemic Brain Damage / 上海针灸杂志

Objective To investigate the effect of different time intervention of Zhu Lian acupuncture exciting method on nerve cell apoptosis and tissue expressions of PI3K (phosphatidylinositol 3-kinase), AKt (serine/threonine kinase) and Caspase-3 (cysteinyl aspartate specific proteinase-3) proteins in young rats with hypoxic-ischemic brain damage.Methods Fifty 7-day-old rats were randomized into groups A (acupuncture exciting methodⅠ), B (acupuncture exciting methodⅡ), C (model), D (sham operation) and E (normal control), 10 rats each. Groups A and E began to receive acupuncture in 24 hours after model making and group B, at 8 days after model making. Groups C and D were not given acupuncture. Every group of animals was sacrificed at 21 days after model making. Nerve cell apoptosis was examined using In situ end labeling (TUNEL) technique. Cerebral expressions of PI3K, AKt and Caspase-3 proteins were determined by immunohistochemical method.Results The number of apoptotic cells was significantly smaller in groups A and B than in group C (P<0.01,P<0.05) and decreased significantly in group A compared with group B (P<0.05). The expressions of PI3K and AKt proteins increased significantly and the expression of Caspase-3 protein decreased significantly in groups A and B compared with group C; there were statistically significant differences (P<0.01,P<0.05). PI3K expression increased significantly and Caspase-3 protein expression decreased significantly in group A compared with group B (P<0.05); there were statistically significant differences (P<0.05).Conclusions Zhu Lian acupuncture exciting method can inhibit nerve cell apoptosis, stimulate the expression of PI3K/AKt signaling pathway, increase PI3K and AKt activities and reduce the expression of Caspase-3 protein in young rats with hypoxic-ischemic brain damage. Early intervention of Zhu Lian acupuncture exciting method is of important significance in producing a protective effect on brain nerves in young rats with hypoxic-ischemic brain damage.

Serum vaspin levels in type 2 diabetic patients are associated with the severity of lower limb vascular lesion / 中国医师杂志

Objective To explore the association between serum vaspin levels and the severity of the lower extremity vascular lesions in type 2 diabetes mellitus (T2DM) patients.Methods According to the lower extremity artery plaque,stenosis and intima-media membrane thickening severity score,92 cases of T2DM patients were divided into three groups:simple diabetes group (DM1 group) (32 cases),mild diabetic lower limb vascular lesion group (DM2 group) (33 cases),and moderately severe diabetic lower limb vascular lesion group (DM3 group) (27 cases).Twenty-six age-and gender-matched apparently healthy controls (control group) were recruited as well.Systolic blood pressure (SBP),ankle-brachial index (ABI),insulin resistance index (HOMA-IR),and other indicators were determined,serum vaspin was measured by enzyme-linked immunosorbent assay.Results After adjustment for age and gender,the DM1 group had a significantly higher serum vaspin level than control group (P ＜0.01),while for the DM2 and DM3 groups,serum vaspin levels were significantly lower than the DM1 group (P ＜ 0.01).Partial correlation analysis showed that serum vaspin was inversely associated with HOMA-IR (r =-0.461,P =0.001)and positively correlated with ABI (r =0.462,P =0.001).Logistic regression analysis demonstrated that SBP,ABI and fasting serum vaspin levels were significantly associated with the presence of the lower extremity vascular lesions in type 2 diabetes.Conclusions Serum vaspin levels were significantly elevated in simple type 2 diabetic patients while the reduction of its level might be associated with the formation of lower extremity vascular lesions in type 2 diabetic patients.

The status of Niger children receiving acupuncture treatment / 国际中医中药杂志

To Describe the current situation of Niger children receiving acupuncture treatment.Acupuncture has 30 years of development history in Niger.Chinese medical teams had established acupuncture department in the Central Hospital in Maradi,Niamey National Hospital,and National Hospital of Zinder,and presided over acupuncture work.But due to a variety of reasons,at present,only the Central Hospital in Maradi has reserved acupuncture therapy,and was combined with physical therapy,rehabilitation therapy to form exercise therapy and functional training division.Pediatric patients are the major subjects who received acupuncture,accounting for 2/3 of outpatients.Nervous system and motor system diseases were their main diseases.

Study on CLCNKB gene mutation in a late-onset Chinese patient with classic Bartter syndrome / 中华肾脏病杂志

Objective To investigate the underlying mutation in a late-onset Chinese patient with classic Bartter syndrome. Methods The mutation analysis of CLCNKB gene was performed by the PCR direct sequencing. The patient's parents and siblings were studied as well. Fifty normal volunteers were analyzed as control group. Results The heterozygous deletion mutation cDNA 753delG and heterozygous missense mutation G433E were detected in the patient. Her father was found to carry heterozygous G433E and her mother to carry cDNA 753delG mutation respectively. Her brother carried heterozygous G433E and her sister was normal. Conclusions Two mutations of the CLCNKB gene in this Chinese patient with late-onset classic Bartter syndrome are identified. The cDNA 753delG mutation has not been reported previously.

Evaluation of alpha 5 (Ⅳ) collagen chain of skin basement membrane in diagnosis of Alport syndrome / 中华肾脏病杂志

Objective To evaluate the detection of alpha 5 (Ⅳ) collagen chain of skin basement membrane in diagnosis of Alport syndrome among suspected patients. Methods Data of suspected patients with the detection of alpha 5 (Ⅳ) collagen chain of skin basement membrane were retrospectively collected and analyzed from January 2007 to March 2008. Results A total of 254 suspected patients ranged from 1 to 71 years old with an average age of (25.85±17.03) years old were enrolled (male/female ratio, 0.76). There was no significant difference in average age between male and female. Abnormal alpha 5 (Ⅳ) collagen chain expression of skin basement membrane was found by indirect immunofluorescense in 19 patients among whom 12 cases were negative and 7 cases were diseontinous deposit. These 19 patients were diagnosed as Alport syndrome and the diagnostic rate was 7.5%. Conclusions The diagnostic rate of Alport syndrome by detection of alpha 5 (Ⅳ) collagen chain in skin basement membrane is significant and helpful for early and differential diagnosis of Alport syndrome.

Mutations of ACTN4 and SYNPO genes promoter in primary focal segmental glomerulosclerosis / 中华肾脏病杂志

Objective To investigate the mutations ACTN4 and SYNPO genes promoter in sporadic primary focal segmental glomerulosclerosis (FSGS) and to analyze the role of mutations in FSGS. Methods The study consisted of 82 Chinese primary FSGS, including 39 females and 43 males, ranged from 12 to 76 years old. Seventy volunteers were selected as healthy control group. Genomie DNA was extracted from peripheral blood cells of FSGS patients and hair of patients' parents by polymerase chain reaction (PCR) and direct sequencing to analyze ACTN4 and SYNPO gene promoter mutations. Mutations were matched with GenBank and TRANSFAC software database (www.ncbi.nlm.nih.gov; www.genometix.de; www.gene-regulation, corn). Dual luciferase assay system was used to analyze the promoter region mutations, based on PGL3-Basie vector, pRL-SV40 and PCI2 cell line. Hair DNA of novel mutation patients' parents was sequenced. Expression of alpha-actinin-4 and synaptopodin in patients' kidney tissue was examined by immunofluorescence. Results Three patients with 1-34C>T, 1-590delA and (1-1044delT)+ (I-797T >C) +(1-769A >G) heterozygous mutations were found in ACTN4 gene promoter respectively, and two patients with 1-24G>A and 1-851C>T heterozygous mutations in SYNPO gene promoter respectively. The same mutations were not found in the control group of 70 healthy people. Except one patient accepting her parents' 1-1044delT and 1-797T>C mutated chromosome respectively, no same mutations were found in patients' parents. Protein expression of alpha-actinin-4 and synaptopodin was reduced in mutated patients' kidneys. Except 1-1044delT group, luciferase activity in mutated groups decreased. (1-1044delT)+(1-797T>C)+(1-769A>G) mutation was associated with poor outcome and patient with these mutations progressed to end-stage renal failure. Conclusion Mutations of ACTN4 and SYNPO gene promoters affect gene transcription and protein translation, which may contribute to the onset of sporadic primary FSGS.

Correlation analysis between hypergammaglobulinemia and renal involvement in patients with primary Sj(o)gren's syndrome / 中华风湿病学杂志

Objective To identify the correlation between hypergnmmaglobulinemia (hyper-IgG) and renal involvement in patients with primary Sjogren's syndrome (Pss). Methods The data of all patients admitted to hospital with the diagnosis of Pss were retrospectively analyzed. One way ANOVA and Speannan's correlation analysis were used to compare the clinical characteristics, renal injuries, immunology tests and renal pathological changes between patients with or without hyper-lgG. Results One hundred and thirty Pss cases were enrolled including 8 males and 122 females. Their age ranged from 16 to 68 years with an average of (44±12) years. Forty-one patients with Pss underwent renal biopsy. The preys lenee of Drta and tubular protein was significantly higher in patients with hyper-IgG than those without(P<0.05). Spearman's correlation analysis showed a negative correlation between serum IgG levels and seral potassium level(r=-0.269,P<0.01).Protein electrophoresis results revealed predominantly tubular protein in the hyper-lgG group, on the other hand glomerular protein was found in the normal-IgG group (P<0.05). The occurrence of decreased C4 com-plement concentration was significantly higher in normal-lgG group (P<0.05); Spearman's correlation analysis for biopsied materials showed that there was no significant difference in the Tubular Index (TI) and Glomeru-lar Index(GI) between these two groups. Conclusion Tubular lesions, especially Drta, may be predominant and correlate with hypergammaglobulinemia. There is a correlation between hypergammaglo-bulinemia and the level of renal lesions. Renal acidification capacity in patients with hypergammaglobulinemia should be evaluated.

Clinicopathological analysis on glomerulonephritis complicating with acute interstitial nephritis / 中华肾脏病杂志

Objective To identify the clinical characteristics and pathological changes of patients suffered from glomerulonephritis complicating with acute interstitial nephritis (AIN) . Methods Twenty one patients of glomerulonephritis complicating with AIN diagnosed by renal biopsy were retrospectively analyzed . Thirty-five pure AIN patients were selected as controls .Results Glomerulonephritis complicating AIN accounted for 37 .5% of all the AIN cases . Βlactam antibiotics and Chinese herbs were the major causes of AIN . 76 .2% of cases received further examinations due to the elevation of serum creatinine (Scr) during their follow-up of kidney injuries or during routine exams for all kinds of discomforts . Pathological features of AIN were also detected besides glomerular leisions . The impairments of renal interstitia were severe than those of the glomeruli . Eosinophil in the renal interstitia was an important indicator for the diagnosis of AIN .The renal function returned to normal or baseline in 64 .7% of the patients of glomerulonephritis complicating with AIN whose follow-up data were available . The median period for renal function restoration was 150 days (compared with 60 days in pure AIN) . But there were no significant differences between these two groups as for the rate of irreversible renal insufficiency during a follow-up period of 2 years . Conclusions Symptoms of AIN in patients of glomerulonephritis complicating with AIN tend to be masked by their glomerular diseases . Renal biopsy is of most importance for the diagnosis . Early diagnosis and treatment leads to satisfactory prognosis .

Mutations of podocyte molecules in patients with late onset familial focal segmental glomerular sclerosis / 中华肾脏病杂志

Objective To investigate the mutations of pedocyte molecules in patients with late onset familial focal segmental glomerular sclerosis (FSGS). Methods Thirty-one pedigrees of late onset familial FSGS in Department of Nephrology, Shanghai Ruijin Hospital from Sep 1997 to Oct 2007 were enrolled in this study. The diagnosis standard of familial FSGS was as follows:(1) the age of presentation was more than 12 years old. (2) in one pedigree, two or more individuals were proven as FSGS by renal biopsy, or at least one was proven to be FSGS by renal biopsy, the others presented renal insufficiency or pmteinuria without precise causes. One hundred unrelated healthy people were screened as control group. Genomic DNA extracted from peripheral blood cells were amplified by PCR and then sequenced for mutations of NPHS2, ACTN4 and TRPC6. Results A novel missense heterozygotic mutation L316P of ACTN4 was identified inone pedigree. The mean onset age of the affected members of this pedigree was (38.7±7.4) years old and their kidney injury progress was slow. Proteinuria of the proband's brother was not improved by immunosuppressor. All 3 affected members of this family had such heterozygotic mutation. A novel missense heterozygotic mutation Q889K of TRPC6 was found in another pedigree. The mean onset age of the affected members in this pedigree was (38.0±4.2) years old. Three members presenting renal disease in this family all had such heterozygotic mutation but with different clinical manifestations. A quiescent mutation G467G of TRPC6 was also identified. Above variants were not found in healthy controls. No NPHS2 mutation was found to cause familial FSGS in these pedigrees. Conclusions A novel mutation L316P of ACTN4 and a new mutation Q889K of TRPC6 are identified in Chinese patients of late onset familial FSGS. No NPHS2 mutation is found to induce FSGS in these pedigrees.

Detection of mutations in 30 exons of COL4A5 gene from X-Iinked Alport syndrome patients / 中华肾脏病杂志

Objective To screen the mutations in 30 exons of COL4A5 gene from X-linked Alport syndrome. Methods 20 Chinese X-linked Alport syndrome patients from 16 families were examined. Genomic DNA was extracted from peripheral leukocyte and exon-specific primers were designed for 30 exons(1-25、31、 32、41、 50、51) . Polymerase chain reaction amplification was performed and followed bydegenerating gel gradient electrophoresis (DGGE) analysis. All abnormal migration bands were sequenced and one hundred normal persons selected as control. Results Four abnormal bands were detected, which were all point mutations and predicted to be functionally pathogenic, in four unrelated patients. One patienthad a nonsense mutation in exon 1 (Glu 22 Term); one had a missense mutation in exon 3l(Gly852Val).Two patients carried splicing mutations in intron 1 and 25 respectively(283+1G→T、2150 + 1G→T) .Conclusions X-linked Alport syndrome is caused by various kinds of COL4A5 gene mutations without any hotspot. Paralleled with the significance of exon mutations, intron mutations also play a critical role in the pathogenesis. Furthermore, these four pathogenic mutations have never been reported in the genebank and showed good correlation with clinical manifestations.

A clinicopathological study of Alport syndrome and detection of type Ⅳ collagen chains in Alport patients / 中华肾脏病杂志

Objective To analyse the clinicopathological changes of Alport syndrome (AS) and to detect the deposition of type Ⅳ collagen within basement membrane of Alport patients. Methods Fourteen patients with AS (12 families) hospitalized from 1990. 1 to 1996. 6 were investigated. Eleven were male and 3 female (mean age 29. 4 years). Results Microscopic hematuria was found in 13, with recurrent gross hematuria in 7. All had proteinuria. Three patients presented nephrotic syndrome. Progressive renal failure occured in 10 of 11 male(11 - 39 years) and 1 female (40 years). Six patients were treated with hemodialysis, two of them with transplantation. Sensorineural deafness was observed in 9 patients particularly high frequency sound. Anterior lenticonus were presented in 2. It showed heterogeneitic, 50% transmitted as X-linked dominant(XD) trait. In 7 renal biopsies, the findings by light microscopy mostly revealed focal and segmental sclerosis glomerulonephritis (4/7). The results of immunofluorescenc e (IF) were ne gatt ye in 4. Ultrastructural studies showed variable thickening, thinning of glomerular basement membrane (GBM) in 7 specimens with lamellation and basket-weaving of GBM in 1. Using the iIF technique, the ?3, 4, 5 (Ⅳ) chains were absent within both GBM and EBM of 4 male XD-AS patients. Conclusions AS is not a rare hereditary disease characterized by hematuria, proteinuria and progressive renal failure with sensorineural deafness and ocular lesions. Type Ⅳ collagen within Alpori patients' basement membrane is abnormal and iIF study of type Ⅳ collagen chains distribution is useful to confirm the diagnosis of AS.