Postnatally confirmed COL4A1 mutation in a fetus with recurrent intraparenchymal hemorrhage and cystic leukomalacia / 中华围产医学杂志

We report a fetus with recurrent intraparenchymal hemorrhage and cystic leukomalacia during pregnancy who was postnatally detected with a de novo mutation in the COL4A1 gene by genetic testing of umbilical cord blood. Multiple fresh hemorrhagic foci were detected in the fetal brain parenchyma and cerebellar hemisphere by ultrasound at 25 gestational weeks. Regular re-examination of the nervous system's ultrasound and magnetic resonance imaging (MRI) indicated recurrent multiple intraparenchymal hemorrhages followed by cystic leukomalacia. However, karyotyping and chromosomal microarray analysis of amniotic fluid showed no abnormality. The newborn was born by cesarean section at 37 +3 gestational weeks with an Apgar score of 10 at 1 and 5 min. Repeated apnea occurred after birth. MRI detected new intraparenchymal hemorrhage and cystic leukomalacia on the six-day of life. The infant's limb muscle tone remained low on the 90-day follow-up. The patient was lost to follow up. Whole-exome sequencing of the cord blood identified a de novo heterozygous mutation- c.4738G>A in the COL4A1 gene (NM_001845.4; p.G1580S) neither parent carried. It suggests that the genetic test of the COL4A1 mutation should be considered for fetuses with intracranial hemorrhage in the prenatal diagnosis, especially those with recurrent fetal intraparenchymal hemorrhage followed by cystic leukomalacia. Genetic tests could help analyze the fetal prognosis, and guide the delivery mode.

Effects of liraglutide on the transforming growth factor-β1/Smads signaling pathway in renal tissues of elderly type 2 diabetic rats / 中华老年医学杂志

Objective:To examine the effects of liraglutide on the transforming growth factor-β1(TGF-β1)/Smads signaling pathway in renal tissues of elderly rats with type 2 diabetes mellitus(T2DM)and to explore the underlying mechanisms.Methods:A total of 75 healthy elderly male Sprague-Dawley rats aged 20 months and weighing(500±100)g were divided into the normal control group(Group N, n=25)and the model group( n=50)by using a random number table.Rats in the model group were given high-glucose and high-fat diets for 8 weeks and then were injected with a single dose(30 mg/kg)of 1% streptozotocin into the abdominal cavity.Forty-eight rats in the model group were successfully molded and were divided into the T2DM group(Group D, n=24)and the intervention group(Group LD, n=24). Rats in Group LD were abdominally injected with liraglutide in a dose of 200 μg·kg -1·d -1, and the other two groups were given an equal volume of saline.At the end of 4, 8 and 12 weeks, eight rats in each group were randomly selected and 24-hour urine collections were made to measure 24-hour urinary protein.Then the rats were anesthetized, blood samples were collected for biochemical tests, and renal tissues were removed for microscopic examination of pathological changes after HE staining.The expression of type Ⅳ collagen(Col-Ⅳ)was detected by using an immunohistochemical method, and the mRNA expression of TGF-β1, Smad3 and Smad7 was detected by using real-time fluorescence quantitative polymerase chain reaction.One-way analysis of variance was used for comparisons between the groups, and the LSD-t test was used for pairwise comparisons. Results:Compared with Group N, Group D showed thickening of the glomerular basement membrane, mesangial proliferation and interstitial fibrosis at each time-point, which grew worse with time, and the expression of TGF-β1 mRNA, Smad3 mRNA and Col-Ⅳ also increased significantly(12-week: 0.69±0.01 vs.0.15±0.01, 0.51±0.02 vs.0.02±0.01, 183.33±2.08 vs.221.67±2.08, t=89.22, 60.87 and 24.52, P<0.05), while Smad7 mRNA levels decreased( t=13.42, P<0.05). Compared with Group D, the degree of renal fibrosis was reduced, and the expression of TGF-β1 mRNA, Smad3 mRNA and Col-Ⅳ at 12-week significantly decreased( t=71.703, 37.58 and 20.04, P<0.05), while Smad7 mRNA increased( t=9.96, P<0.05)in Group LD.With prolonged intervention of liraglutide, the lesions were mitigated, the expression of TGF-β1 mRNA, Smad3 mRNA and Col-Ⅳ decreased, and Smad7 mRNA increased gradually( P<0.05)in Group LD. Conclusions:Liraglutide has anti-renal fibrosis effects via inhibiting the TGF-β1/Smads pathway, thereby reducing the production of Col-Ⅳ, and can delay the progression of renal lesions in elderly T2DM rats.

Clinical value of biochemical indicator and hepatic fibrosis in differential diagnosis of patients with cirrhosis and chronic liver failure / 中国基层医药

Objective@#To investigate the clinical value of biochemical indicator and hepatic fibrosis in differential diagnosis of patients with cirrhosis and chronic liver failure.@*Methods@#From December 2015 to December 2018, 30 patients with cirrhosis and 30 patients with chronic liver failure in Zhoushan Hospital were selected.The serum levels of ALT, AST, TBil, hyaluronic acid (HA), laminin (LN), PC-Ⅲ, collagen type Ⅳ(Ⅳ-C) were detected.@*Results@#The serum levels of ALT, AST and TBil in patients with cirrhosis were significantly lower than those in patients with chronic liver failure[(258.17±88.19)U/L vs.(818.37±375.83)U/L; (256.57±97.21)U/L vs.(738.63±329.68)U/L; (51.37±22.13)μmol/L vs.(157.27±85.60)μmol/L, t=-7.95, -7.68, -6.56, all P<0.01]. The serum levels of HA, LN, PC-Ⅲ and Ⅳ-C in patients with cirrhosis were significantly higher than those in patients with chronic liver failure[(433.10±214.39)μg/L vs.(81.56±27.80)μg/L; (157.67±52.60)μg/L vs.(66.06±24.06)μg/L; (126.63±35.24)μg/L vs.(70.96±19.71)μg/L; (162.26±53.93)μg/L vs.(76.13±27.90)μg/L, t=8.90, 8.67, 7.49, 7.76, all P<0.01]. When patients with chronic liver failure were set as state variable, AUC of ALT(0.973) was the highest in ALT, AST, TBil.When patients with cirrhosis were set as state variable, AUC of HA(0.993) was the highest in HA, LN, PC-Ⅲ and Ⅳ-C.@*Conclusion@#ALT and HA are suitable for differential diagnosis of patients with cirrhosis and chronic liver failure.

A family of Alport syndrome with a novel dominant hereditary hematuria and a mutation in the COL4A4 gene / 中华检验医学杂志

Objective@#Alport syndrome was an inherited kidney disease caused by the mutation of COL4A3, COL4A4, or COL4A5. Whole-exome sequencing was used to detect the mutations on these genes for the molecular diagnosis of Alport syndrome.@*Methods@#A 6-year-old girl found accidentally with microscopic hematuria at the age of 4. The clinical data and blood sample of the family including proband, parents, brothers, and sisters were collected. Whole exome sequencing was conducted using their genomic DNAs.@*Results@#A novel heterozygous frameshift mutation c.1826delC (p.Pro609Glnfs*44) was found in the exon 25 of the COL4A4 (NM_000092) in the proband, the father, and the sister, showing an autosomal dominant inheritance pattern of Alport syndrome. This mutation of COL4A4 was confirmed by mutation analysis, and the mutation of c.1826delC was verified by Sanger sequencing. No mutations on COL4A3 and COL4A5 were detected in this family. And the mother and brother are normal wide-type.@*Conclusions@#This novel mutation is a valuable addition to the current genetic profile of Alport syndrome, and provide us a better understanding of the disease. Whole-exome sequencing is a power tool to identify the novel mutations of inherited disease and contribute to the molecular diagnosis of disease.

A family of Alport syndrome with a novel dominant hereditary hematuria and a mutation in the COL4A4 gene / 中华检验医学杂志

Objective Alport syndrome was an inherited kidney disease caused by the mutation of COL4A3,COL4A4, or COL4A5. Whole-exome sequencing was used to detect the mutations on these genes for the molecular diagnosis of Alport syndrome. Methods A 6-year-old girl found accidentally with microscopic hematuria at the age of 4. The clinical data and blood sample of the family including proband, parents, brothers, and sisters were collected. Whole exome sequencing was conducted using their genomic DNAs. Results A novel heterozygous frameshift mutation c.1826delC (p.Pro609Glnfs*44) was found in the exon 25 of the COL4A4(NM_000092) in the proband, the father, and the sister, showing an autosomal dominant inheritance pattern of Alport syndrome. This mutation of COL4A4 was confirmed by mutation analysis, and the mutation of c.1826delC was verified by Sanger sequencing. No mutations on COL4A3 and COL4A5 were detected in this family. And the mother and brother are normal wide-type. Conclusions This novel mutation is a valuable addition to the current genetic profile of Alport syndrome, and provide us a better understanding of the disease. Whole-exome sequencing is a power tool to identify the novel mutations of inherited disease and contribute to the molecular diagnosis of disease.

Construction of Dual-luciferase reporter gene vector of 3'UTR of the COL4A3 gene and validation of its targeting relationship with miR-299 / 中华显微外科杂志

Objective To construct a dual-luciferase reporter gene vector and validate the targeting relation ship between miR-299 and the COL4A3 gene,laying a foundation for the study on the effect of miR-299 in the chondrogenic differentiation of stem cells by regulating the COL4A3 gene.Methods This study was made from March,2018 to December,2018.Firstly,the potential binding sites between miR-299 and COL4A3-3'UTR were pre dicted using bioinformatics.Then,the wild and mutant COL4A3-3'UTR sequences were amplified by PCR and cloned into psiCHECK-2 plasmid to construct corresponding recombinant vectors.The vectors were validated by enzyme digestion and gene sequencing.Finally,the cells were resuscitated,amplified,transfected and divided into 4 groups:COL4A3-WT+miR-299/NC group,COL4A3-WT+miR-299-inhibitor/NC-inhibitor group,COL4A3-MUT+miR-299/NC group and COL4A3-MUT+miR-299-inhibitor/NC-inhibitor group.Each group contains 3 holes,respectively.Luciferase activity in each group was determined using a dual-luciferase assay kit.The statistical analysis was conducted and differences between groups were compared by t test.Probabilities lower than 5％(P＜0.05) were considered statistically significant.Results Enzyme digestion and DNA sequencing showed that the dual-luciferase reporter gene vector of psiCHECK-2-COL4A3 was constructed successfully.Luciferase assay demonstrated that in wild COL4A3 gene,luciferase activity reduced in the miR-299 transfection group (The average R/F value was 59.38％) compared with the NC group (The average R/F value was 100.00％),with a statistical significant difference (P＜0.05).In wild COL4A3 gene treated with inhibitor,luciferase activity increased in the miR-299-inhibitor group (The average R/F value was 153.98％) compared with the NC-inhibitor group (The average R/F value was 100.00％),with a statistical significant difference (P＜0.05).In mutant COL4A3 gene treated with inhibitor,no obvious statistical differences in luciferase activity were found between miR-299 transfection group (The average R/F value was 102.09％),miR-299-inhibitor group (The average R/F value was 108.51％) and NC group (The average R/F value was 104.70％),NC-inhibitor group (The average R/F value was 105.13％) and/9＞0.05.Conclusion The dual-luciferase reporter gene vector of the 3'UTR of the COL4A3 gene is constructed successfully.In addition,dual-luciferase assay further verifies the authenticity of miR-299 directly targeting the 3'UTR of the COL4A3 gene.

Influence of angiogenin-like protein 4 on pulmonary fibrosis / 中国医师杂志

Objective To investigate the important role of angiogenin-like protein 4 (Angptl4) in pulmonary fibrosis and to provide a new therapeutic targets for pulmonary fibrosis.Methods We established the pulmonary fibrosis animal models in rat by tracheal instillation of bleomycin.Then,we detected the expression of Angptl4 through real-time polymerase chain reaction (RT-PCR) and Western Blot.Rat lung fibroblast (RLF) was transfected into Angptl4-shRNA plasmid.Then we detected the changed collagen expression in RLF cells after transfection through RT-PCR and Western blot.Results The expression of Angptl4 was up-regulated in the bleomycin-induced rat pulmonary fibrosis model.The expression of both collagen Ⅰ and collagen Ⅳ in RLF cells transfected with Angptl4-shRNA plasmids were down-regulated compared with control after TGF-β treatment.Conclusions Inhibiting the expression of Angptl4 can reduce the expression of collagen fibers in lung tissue,then delaying the progression of pulmonary fibrosis.

Effect of genistein on deposition of extracellular matrix in uremic rats / 重庆医学

Objective To investigate the effect of genistein on deposition of extracellular matrix(ECM) in uremic rats and its mechanism.Methods The uremic rat model was established by 5/6 nephrectomy.The model rats were divided into the genistein group(G) and control group (C).The sham-operation group served as the normal control(S).Urinary protein and biochemical indexes before operation,at postoperative 4,8 weeks were measured in various groups.The pathologic changes of renal tissue were observed.The glomerular sclerosis index(GSI) was evaluated by the semi-quantitative method.fibronectin(FN) and type Ⅳ collagen (ColⅣ) deposition situation were detected by using the immunohistochemical method.Western blot and RT-PCR were used to measure the protein expression and mRNA transcription of TGF-β1.Results Comparing postoperative 4 weeks with the group C,the urinary protein excretion amount was decreased[(11.63 ± 2.07) mg/d vs.(19.93 ± 3.19) mg/d,all P＜ 0.01],serum urea nitrogen and creatinine levels of the group G were decreased[(9.39±0.59)mmol/L vs.(12.09±0.78)mmol/L,(65.11±3.79)mmol/L vs.(77.63±3.20)μmol/L,all P＜0.01].Until postoperative 8 weeks,the urinary protein excretion amount was decreased and the decrease of serum urea nitrogen and creatinine levels was more obvious.The deposition of ColⅣ and FN in renal glomerulus was lower than that of the control group[(17.30±1.96)％ vs.(24.68±3.97)％;(18.26±2.31)％ vs.(29.35±4.15)％,all P＜ 0.01].Glomerular sclerosis was significantly alleviated.The TGF-β1mRNA and protein expression were attenuated (P＜0.05).Conclusion Genistein can reduce the deposition of ECM in uremic rats and has a protective effect on kidney.

The effect of daidzein on extracellular matrix of uremic rats / 国际中医中药杂志

Objective To observe the effect of daidzein on extracellular matrix of uremic rats and to discuss its mechanism.Methods Uremic rat model were established by 5/6 nephrectomized. Model rats were devided into daidzein group, control group. Rats with sham-operation were regarded as the normal control. At time of baseline, 4th and 8th week after operation, urinary protein and biochemical detection were measured. The pathologic changes, fibronectin (FN) and typeⅣcollagen (ColⅣ) were investigated at 8th week. The Western-Blot and RT-PCR were used to measure protein expression and mRNA transcription of TGF-β1. Results At 8th week after operation, the urinary protein (12.35 ± 2.13 mg/24 hvs. 19.93 ± 3.19 mg/24 h), serum urea (10.11 ± 0.65 mmol/Lvs.12.09 ± 0.78 mmol/L) and creatinine (68.10 ± 2.51μmol/Lvs.77.63 ± 3.20μmol/L) in the daidzein group decreased significantly than those in the control group (P<0.01). The deposition of ColⅣ (16.33% ± 2.14%vs. 24.68% ± 3.97%) and FN (19.17 ± 2.68 vs. 29.35 ± 4.15) in the daidzein group decreased significantly than those in the control group (P<0.01). Compared with control group, the pathological lesion in the daidzein group was less serious. The mRNA transcription (0.37 ± 0.06vs. 0.64 ± 0.08) and protein expression of TGF-β1 (0.28 ± 0.09vs. 1.40 ± 0.13) in the daidzein group were attenuated significantly than those in the control group (P<0.05). ConclusionsDaidzein had a beneficial effect on uremic rats. It may be associated with a decrease of extracellular matrix accumulation.

Identification of a novel COL4A4 mutation in a family with thin-basement-membrane nephropathy and the pathogenic mechanism / 中华肾脏病杂志

Objective To explore a new pathogenic gene mutationin in COL4A3 and COL4A4 genes of a family with thin-basement-membrane nephropathy (TBMN), and explain its mechanism.Methods Genomic DNA was extracted from blood samples.Mutation screening for all the exons in COL4A3 and COL4A4 of the proband was carried out by direct PCR sequencing.The sequences of the proband were compared with standard sequences in GenBank.After identifying the mutation in COL4A4, screening for the mutation site in 200 healthy controls and the rest of family members were conducted.RNA sequence of the proband was analyzed by reverse transcription PCR and TA cloning.The positive clones were sequenced for RNA screening.Results There was a G to A mutation in the 1459 site of COL4A4 (c.1459+G ＞ A) in the proband, her mother, and the elder sister, whereas the mutation was not found in other family members and healthy people.RNA screening showed that the COL4A4 (c.1459+G ＞ A) mutation was a heterozygous substitution in position + 1 of exon 21, in the splicing region.This mutation leaded to eliminating of exon 21 from the COL4A4 mRNA, causing the exon 21 deletion and frameshift mutation following the exon 20 in its amino acids sequence.Conclusions It is described that COL4A4 (c.1459+G ＞ A) is a new pathogenic mutation in TBMN, which further help understanding the pathogenesis and clinical diagnosis of TBMN.

Study of the effect and mechanism of relaxin on the ECM excretion of Human renal mesangial cells cultured in high ambient glucose / 中国医师杂志

Objective To explore the effect and mechanism of relaxin on the production of extracellular matrix (ECM) excreted by high glucose stimulated human renal mesangial cells.Methods Cultured human mesangial cells (HMCs) were divided into three groups:(1) normal glucose group (NG,5.5 mmol/L D-glucose),(2) high glucose group (HG,30 mmol/L D-glucose),and (3) high glucose + relaxin group.Cell count kit (CCK8) was used to examine the cell proliferation.The levels of fibronectin and collagen type Ⅳ in the culture supernatants were examined with a solid-phase enzyme-linked immunoadsorbent assay (ELISA);Western blot method was used to detect the expression of α-smooth muscle actin (α-SMA) protein.The transforming growth factor-β1 (TGF-β1) mRNA expression was detected with quantitative polymerase chain reaction (qPCR) method.Results No proliferation and inhibition effects were observed in both normal and high glucose group.Compared to the normal glucose group,the levels of fibronectin,and collagen type Ⅳ increased significantly (57.28 ± 0.59 vs 41.85 ± 0.03,56.52 ± 0.88 vs 33.80 ± 0.24,P ＜ 0.01)after cultured 48 h in high concentration of glucose.Compared to the high glucose group,a significantly decreases of fibronectin and collagen type Ⅳ (47.08 ± 0.03 vs 57.28 ± 0.59,36.16 ± 0.52 vs 56.52 ±0.88,P ＜0.01) were observed in the relaxin treated group.The expressions of α-smooth muscle actin and TGF-β1 were decreased (P ＜0.01).Conclusions Relaxin can suppress the overproduction of ECM excreted by HMC cultured in high ambient glucose,and its mechanism is partly due to the inhibition of TGF-β1.

The co-expression of human epidermal growth factor 2 and collagen Ⅳ in human breast cancer tissues detected by quantum dots-based immunofluorescence technique and its clinical significance / 中华普通外科杂志

Objective By using quantum dots-based double-color imaging method to simultaneously label human epidermal growth factor 2 (HER2) on breast cancer cell membrane and the type Ⅳ collagen in the extracellular matrix.To monitor the degree of malignancy of breast cancer and the invasive and metastatic potential.Methods The co-expressions of HER2 and the type Ⅳ collagen in breast cancer of 54 cases were detected and quantified by quantum dots-based double-labeling immunofluorescent histochemistry,and its correlation with clinical pathology parameters and prognosis were analyzed.Results With the increasing HER2 expression level,a progressive decrease in collagen Ⅳ around the cancer nest,the expression of HER2 and the type Ⅳ collagen value was negatively correlated (r =-0.980,P ＜ 0.05) ; the expression of HER2 and collagen Ⅳ was associated with lymph note metastasis,pathological stage (TNM) and disease-free survival(P ＜ 0.05).The expression of HER2 and the type Ⅳ collagen between different age groups,menopause and estrogen receptor (ER) and progesterone receptor (PR) were not significantly different (P ＞ 0.05).Conclusions Quantum dots-based double-color imaging method provides direct observable evidence to support the degradation of HER2 and the type Ⅳ collagen expression,which may help in determining the degree of malignancy and evaluating prognosis.

Effect of kistrin on expression of collagen type IV in lens posterior capsule / 中华实验眼科杂志

BackgroundThe proliferation of lens epithelial cellsLECs) following extracapsular cataract extraction is the biological basis of posterior capsular collagen and cataract formation.Disintegrin is certified to competitively bind the integrin with extracellular matrix and therefore prevent the posterior capsular opacification (PCO).But,its molecular mechanism is below understand.ObjectiveThe present study was to investigative the effects of disintegrin (kistrin) on the expression of collagen in lens posterior capsular.MethodsThe right eyes of 24 New Zealand white rabbits received transparent lens extracapsular enudeation and were randomly divided into two groups using random number table,0.2 ml of kistrin ( 80 mg/L) was intracapsularly injected at end of the operation in 12 eyes ( kistrin group) and the same volume of normal saline solution was used at the same way in other 12 operative eyes ( normal saline group).The PCO was graded in postoperative 1,3,5,7,14 days on Odrich' s criteria under the slit lamp.The lens section was prepared at 14 days and 3 months after operation.Haematoxylin and eosin stain was used to examine the proliferation of LECs in posterior capsule; Masson stain was used to observe the collagenous fiber formation in capsule bag,and the expression of collagen type Ⅳ was detected by immunochemistry.Results No significant difference in the number PCO eye was found in postoperative 14 days between normal saline group and kistrin group ( P =0.093 ).However,the eye numbers of 2-3 grades of PCO were significant increased in normal saline group compared with kistrin group in 1,2,3 months after operation( P=0.041,0.014,0.022).In the operative 14 days,staining and adhesion of LECs in posterior capsule were more in normal saline group than kistrin group,and the fibrocytes in capsule were evidently increased in normal saline group in 3 months.Masson stain certified that the blue stain was seen to be stronger and more in posterior capsule in normal saline group in comparison with kistrin group in 3 months after operation,and the immunochemistry showed that the gray values of collagen type Ⅳ in posterior capsule were significant lower in normal saline group compared with kistrin group in both 14 days and 3 months after operation (P=0.000,0.001 ).ConclusionsKistrin can suppresses the proliferation of LECs and collagen type Ⅳ on rabbit lens posterior capsular after transparent lens extracapsular enudeation.

Clinical significance of the serum fibrosis parameters in Chronic hepatitis B patients / 中国基层医药

Objective To study the relationship between serum fibrosis parameters levels and chronic viral hepatitis B patients.Methods Radioimmunoassay and chemiluminescence were used to test the levels of hyaluronic acid(HA),collagen type Ⅳ(Ⅳ-C),laminin(LN) and procollagen type Ⅲ(PC Ⅲ) in 146 cases of chronic hepatitis B and 40 of normal controls,and the relationship with clinical parameters of liver function were analyzed.Results The levels of HA,Ⅳ-C,LN,PC Ⅲ in chronic hepatitis B patients were higher than those of normal control group (P ＜0.05 or P ＜ 0.01),there were statistical differen in different course of disease (P ＜ 0.05 or P ＜ 0.01) ; The levels of serum Ⅳ-C and PCⅢ had positive correlation with plasma PT,the same to HA with TBil and PCⅢ with ALT;The levels of HA,Ⅳ-C and LN had negative correlation with Alb,the same to LN with CHE.Conclusion The levels of HA,Ⅳ-C,LN and PC Ⅲ in chronic hepatitis B patients may reflect the situations of hepatic fibrosis and the degrees of liver function damage.

Changes of serum PCⅢ,Ⅳ-C,LN,TPO and EPO in patients with Leukemia / 中国基层医药

Objective To detect the levels of serum procollagen type Ⅲ (PCⅢ), Ⅳ collagen(Ⅳ-C) ,laminin ( LN ) , erythropoietin ( EPO ) and thrombopoietin ( TPO) in patients with acute and chronic leukemia. Methods PCⅢ , Ⅳ-Cand LN levels were determined by radioimmunoassay and TPO,EPO levels were determined by enzyme-linked immunosorbent assay,for 26 acute leukemia patients,16 chronic leukemia patients,and 16 normal controls. Results PC Ⅲ[(164.02 ± 118.53) μg/L], Ⅳ-C[(66.54 ± 19.48) μg/L] ,LN[(122.85 ±25.10)μg/L], EPO[(52.90±34.87)mU/ml]and TPO[(642.54±318.49)ng/L]levels of patients with acute and chronic leukemia were higher than that of the healthy control group before chemotherapy (P ＜ 0. 05). EPO [( 72. 21 ± 52. 15 )mU/ml]and TPO[(642.54 ±318.49)ng/L]levels of acute leukemia patients after treatment were significantly lower than before treatment(P＜0.05),while PC Ⅲ ,Ⅳ-C,LN levels had no significant difference(P ＞0.05). There was no significant difference before and after chemotherapy for PC Ⅲ, Ⅳ-C,LN,TPO and EPO levels of chronic leukemia patients (P＞ 0.05). Conclusion PC Ⅲ,Ⅳ-C,LN,TPO and EPO of patients with acute and chronic leukemia were abnormal, and dynamic monitoring of EPO and TPO levels could reflect the effect of AL chemotherapy, but conld not contribute to the prediction of CML blast crisis.

Reconstruction of adenovirus vector contained canstatin gene and hTERT gene core promoter and its anti-tumor effect in human ovarian cancer / 中华妇产科杂志

Objective To study anti-tumor effects on human ovarian cancer xenografted tumors in mice by constructing adenoviral expression vector containing canstatin gene and hTERT gene core promoter (AdhTERT-Can). Methods AdhTERT-Can vector was constructed and identified by means of enzyme cutting, electrophoresis and sequencing. Then, transfected into HO8910PM cells by means of lipofectamine and confirmed by green fluorescence protein(GFP) expression under laser confocus microscope. The mRNA expression of canstatin gene was tested by RT-PCR. Human ovarian cancer xenografted tumor models in nude mice were established and randomly divided into AdhTERT-Can group received viral supematant solution of AdhTERT-Can by tail vein injection, Ad-Can groups received viral supernatant solution of Ad-Can by tail vein injection, and control groups received phosphate buffer solution (PBS). The volume of tumors were measured and compared in each group to evaluated the anti-tumor efficacy. Results All the constructed vectors of AdhTERT-Can were verified by enzymed digestion. There were green fluorescence from 60% of HO8910PM cells transfected by AdhTERT-Can under laser confocus microscope, and the mRNA expression of canstafin gene in HO8910PM cells were also verified by RT-PCR The growth of tumor in AdhTERT-Can group was significantly inhibited compared with those in Ad-Can groups and control groups from the 8th day (P <0.01 ). However, there were not significant difference between Ad-Can group and PBS group (P> 0.05). On the 30th day, the tumors showed liquefaction necrosis and cystic degeneration in each group, especially in AdhTERT-Can group. Conclusions The recombinant adenovirus vector of AdhTERT-Can has been constructed successfully and could steady express in ovarian cancer cell lines HO8910PM. The results shown that it could inhibit significantly the growth of human ovarian cancer xenografted tumors in mice and shown to be the target of gene therapy.

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.

Expression and pathobiological significance of ColⅠ,Col Ⅳ and Fn in laryngeal squamous cell carcinomas / 临床耳鼻咽喉头颈外科杂志

Objective:To investigate the expression of collagen typeⅠ(ColⅠ),collagen typeⅣ(ColⅣ)and fibronectin(Fn)in laryngeal squamous cell carcinomas(LSCC)and their pathobiological relationship with invasion and metastasis of tumor.Method:The expression of ColⅠ,Col Ⅳ and Fn was detected by immunohistochemistry method in normal tissue of latero-carcinoma and tissue of carcinoma in 60 specimens of LSCC.Integral optical density(iOD)was detected by image analysis of computer and was analyzed by SPSS13.O.Result:The expression of ColⅠ was obvious and integrity.The expression of Col Ⅳ and Fn of basal membrane was like intact line-shape appearance and Fn of interstitial substance appeared like a complete network in the normal tissue f latero-carcinoma.Their expression decreased gradually and their integrity was broken and disappeared gradually from well to poorly differentiated LSCC.Their expressions also fell off with the tumor size,clinical stage and cervical lymphatic etastasis gradually and consistently in LSCC.Their variances were statistical significance separately(P＜0.05 or P＜0.01).Conclusion:The expression of ColⅠ,ColⅣand Fn was closely related to tumor invasion,the regional lymph node metastasis and other some pathobiological features in LSCC.A detection of ColⅠ,ColⅣ and Fn as of definite significance on a better comprehension of the possibility of metastasis,choice of surgery and prognosis.

Appraisal of determination of serum type Ⅳ collagen protein with latex-enhanced immunoturbidimetry / 国际检验医学杂志

Objective To appraise determination of serum type Ⅳ collagen protein with latex-enhanced immunoturbidimetry.Methods The evaluation test was performed including precision, linearity determination, prozone reaction, correlation, recovery test and interference test.Results The intra-batch CV% was within 2.47%-3.48%, inter-batch CV% was within 3.46%-4.07%, and recovery rate was within 96.2%-99.3%. There was a good linearity, and the tolerance limit was 800 ng/mL.Regression equation:Y=1.011 8X-1.788 3;detection correlation of different concentrations of standard preparations:r2=0.999 9,Y=1.003 1X-1.236 2.The assay had a very good anti-interference performance to bilirubin, cruorin, and triglyceride without exception.Conclusion Determination of serum type Ⅳ collagen protein attains the demand of the clinic with latex-enhanced immunoturbidimetry in fully automatic biochemical analyzer.

Changes of hyaluronic acid,laminin and collagen typeⅣwithin the renal grafts and in serum during acute rejection / 中华器官移植杂志

Objective To study the change of the hyaluronic acid (HA),laminin (LN) and col- lagen type IV (C_(?)) within the renal grafts and in serum during acute rejection and to investigate their relationship.Methods Male Wistar and Sprague-Dawley rats were used as donors and recipients,re- spectively.Rat orthotopic kidney transplantation was performed according to a modification of the method decribed by Biota.Experimental rats were randomly divided into 4 groups,homotransplanta- tion rats treated with cyclosporine,isotransplantation,pseudooperation and controls.Animals were subsequently killed at defined time points for determination of the extracellular matrix (ECM) parame- ters (HA,LN and C_(?)) within the graft and in serum by radioimmunoassay.Results Significant increases in HA,LN and C_(?) levels within the renal grafts were found in the rejection group as compared with the non-rejection groups.Serum levels of HA,LN and C_(?) were also significantly elevated in the rejection group at diagnosis of rejection.Serum HA,LN and C_(?) levels were correla- ted with those within the renal grafts.Histologic examination revealed that 4 cases developed acute re- jection in homotransplantation rats treaded with cyclosporine,17 cases in controls.HA,LN and C_(?) levels within the renal grafts were correlated with acute rejection Banff scores.There was correlation between serum levels of HA,LN and C_(?) and acute rejection Banff scores (P＜0.01).Conclusion HA,LN and C_(?) play an important role in the pathogenesis of acut allograft rejection.In addition, serum levels of HA,LN and C_(?) may be a sensitive marker of acute rejection in the postoperative period of renal transplantation.