Effects of FHL1 on biological character of lung adenocarcinoma cell line A549 / 复旦学报（医学版）

Objective To explore the effect of expression of FHL1 on biological character of lung adenocarcinoma cell line A549.Methods Lung adenocarcinoma cell line A549 was transfected by FHL1 mRNA and shRNA lentivirus vector.The stable overexpression of FHL1 in lung adenocarcinoma cell line A549 model and low expression of cell line model were tested by Western blot and RT-PCR method.CCK-8 test,Transwell invasion,flow cytometry and Colony-forming Assay were used to test the proliferation,invasion,apoptosis and anchorage independence growth of the cell line.Results The difference between the stable overexpression of FHL1 group and its low expression group was obvious tested by Western blot and RT-PCR.CCK-8 test showed higher cell proliferation of the low expression group than that of the overexpression group and the blank group (P =0.002 2).Flow cytometry showed that the apoptosis rate of the overexpression group was 50.48 ％,significantly higher than that of the two low expression groups (16.41％,20.12％) and blank group (25.90％,P＜0.001).The invasiveness of the overexpression group was obviously lower than the low expression group and blank group in the Transwell invasion (P＜0.001).Colony-forming Assay showed that the colony number of the overexpression group was 28.7 ± 6.0,significantly less than the two low expression groups (157.0 ± 6.4,150.7 ± 9.5,P＜0.000 1).Conclusions The stable overexpression and low expression of FHL1 in lung cancer cell line A549 models were successfully constructed.The overexpression of FHL1 inhibited the proliferation,invasion and metastasis,and accelerated apoptosis in lung adenocarcinoma cell line A549.

Expression of FHL1 and the significance in pancreatic fibrosis of chronic pancreatitis / 中华胰腺病杂志

Objective To explore the expression of transcriptional regulation factor FHL 1 in pancreatic fibrosis of chronic pancreatitis (CP) and clinical significance.Methods Serum samples from 20 CP patients treated in the Department of Gastroenterology , Nanjing General Hospital and 20 healthy controls in the corresponding period were collected .The level of FHL1 in the serum was detected by ELISA .Twenty male Wistar rats were randomly divided into control group and CP group according to the body weight .The CP rat model was established by injecting dibutyltin dichloride ( DBTC) through tail vein .All the rats were killed at 28 days after injection and the pancreas was collected , which was observed using hematoxylin and eosin ( HE) staining to investigate pathological changes . The expression of FHL1 in pancreas was measured by immunohistochemistry (IHC) and Western blotting (WB).Results The average of serum FHL1 level in two groups were (0.86 ±0.22)g/L and (1.18 ±0.22)g/L, respectively.The level of FHL1 in serum of CP patients was significantly lower than that of healthy cases , and the difference was statistically significant (t=10.54, P=0.001).Damaged pancreatic lobular structure , acinar atrophy and the obvious fibrous hyperplasia were observed in CP rats .The pathological score of abnormal structure , acinar atrophy, fibrosis, and inflammatory cell infiltration in the pancreas of the CP group was (2.2 ±0.4),(2.0 ±0.7),(2.4 ±0.3) and (2.8 ±0.6), respectively, but was all 0 in control group.The difference between two group was statistically significant ( t value =17.24, 9.01, 25.22 and 14.75, respectively, all P<0.001).The expression of FHL1 in the pancreatic tissue of CP group were less than that of normal group , and the difference was statistically significant (IHC:15 798 ±3 247 vs 37 570 ±9 254, t=2.438,P=0.016;WB:0.44 ±0.11 vs 0.89 ±0.03, t=4.289, P=0.002).Conclusions The protein of FHL1 was in low expression in the pancreas of CPs , indicating that FHL1 may participate in the formation of pancreatic fibrosis .

A novel PRF1 gene mutation in a fatal neonate case with type 2 familial hemophagocytic lymphohistiocytosis / 소아과

Hemophagocytic lymphohistiocytosis (HLH) occurs in the primary form (genetic or familial) or secondary form (acquired). The familial form of HLH (FHL) is a potentially fatal autosomal recessive disorder that occurs because of constitutional defects in cell-mediated cytotoxicity. Here, we report a fatal neonatal case of type 2 FHL (FHL2) that involved a novel frameshift mutation. Clinically, the newborn presented with severe sepsis-like features and required mechanical ventilation and continuous venovenous hemodiafiltration. Flow cytometry analysis showed marked HLH and complete absence of intracytoplasmic perforin expression in cytotoxic cells; therefore, we performed molecular genetic analyses for PRF1 mutations, which showed that the patient had a compound heterozygous mutation in PRF1, that is, c.65delC (p.Pro22Argfs*2) and c.1090_1091delCT (p.Leu364Glufs*93). Clinical and genetic assessments for FHL are required for neonates with refractory fever and progressive multiple organ failure, particularly when there is no evidence of microbiological or metabolic cause.

Novel Mutations in the UNC13D Gene Carried by a Chinese Neonate with Hemophagocytic Lymphohistiocytosis

Hemophagocytic lymphohistiocytosis (HLH) in different ethnicities has been described in the literature, but few cases in patients of Chinese descent have been reported. Here, we describe the case of a Chinese neonate presenting with HLH carrying novel, compound heterozygous mutations of the UNC13D gene, including [c.2295_2298delGCAG, p.Glu765Aspfs*27] in exon 23, c.-250C>T, c.1+30G>A, c.279C>T, c.888G>C, c.18+36A>G, c.20-48T>C, c.1977C>T, c.2296C>T, c.24-46C>T, c.26-9_26-8insC, c.2599A>G, c.28+48C>T and c.3198A>G, some of which have not been reported in the literature. Cytokine profile analyses were performed in this patient, and the results were consistent with our previous findings in HLH patients. Cytokine profile monitoring may be helpful in differentiating among various clinical phases of HLH.

Novel STXBP2 Mutation Causing Familial Hemophagocytic Lymphohistiocytosis.

Familial Hemophagocytic Lymphohistiocytosis (FHL) is a rare autosomal recessive disorder. Diagnosis is established in presence of genetic mutation or positive family history in one of the siblings. Common genetic mutations associated with FHL are mutations in gene PRF-1 (also known as FHL 2), UNC13D (FHL 3) and STX11 (FHL 4). Recently mutation in STXBP2 encoding syntaxin binding protein 2 (Munc 18 -2) has been found to be associated with FHL type 5. Here we describe the first reported Indian patient with homozygous mutation in STX BP2 gene (c1697 G>A resulting in amino acid change p.G566D) causing FHL 5.

Treatment of Old Achilles Tendon Rupture using Modified Flexor Hallucis Longus Tendon Transfer / 대한족부족관절학회지

PURPOSE: The purpose of this study was to evaluate the clinical results of the old Achilles tendon rupture treated with modified flexor hallucis longus (FHL) tendon transfer. MATERIALS AND METHODS: Seventeen patients with old Achilles tendon rupture treated with modified FHL tendon transfer between March 2004 and February 2008 were enrolled in this study. Technically FHL was pass through the distal portion of the ruptured tendon instead of the drilled hole made on the calcaneus. The mean age of the patients was 37 years (range, 22~67 years), mean follow-up period was 28 months (range, 12~30 months). Patients' subjective satisfaction, calf circumferential diameter, range of motion of ankle and AOFAS ankle-hind foot score and Arner-Lidholm score was evaluated. RESULTS: The average gap between the ruptured tendon was 52 mm (range, 47~56 mm). The AOFAS score improved from 47 pre-operatively to 91 points at the last follow-up. Sixteen patients were satisfied with the result free from discomfort, a patient had mild discomfort who had DM. fourteen patients had decreased range of motion less than 5 degrees while 2 patients had more than 7 degrees decrease compared to the intact side but had no discomfort in daily activities. Nine patients had less than 1 cm calf circumferential diameter difference and 7 patients had 1 to 3 cm diameter difference compared to the intact side. One who had more than 3 cm diameter difference had deteriorated muscle strength. CONCLUSION: Modified FHL tendon transfer can be a useful technique for the treatment of old Achilles tendon rupture when the gap is with large gap placed too proximal.

Inhibitory effect of FHL2 on Id1-mediated suppression of proliferation and invasive capacity of breast cancer cells MCF-7 / 解放军医学杂志

Objective To investigate the inhibitory effect of four and a half LIM protein 2(FHL2) on inhibitor of differentiation 1(Id1)-mediated suppression of transcriptional regulation activity and Id1-promoted invasive growth of human breast cancer cells MCF-7.Methods The effect of FHL2 on Id1-mediated transcriptional repression in MCF-7 cells was determined by cotransfection and relative biluciferase assay.The cell proliferation was determined by MTT assay,and the invasive capacity of MCF-7 cells was determined by Transwell assay.Results The transcriptional repression effect of Id1 on basic helix-loop-helix(bHLH) factor E47-mediated transcription activity in MCF-7 cells,and Id1-promoted proliferation and invasive growth of MCF-7 cells were significantly suppressed by FHL2.Conclusion FHL2 can inhibit the proliferation and invasive growth of human breast cancer cells via repressing Id1-mediated functional activity.The results provide a basis for further investigating the functional roles of FHL2-Id1 signal pathway in the carcinogenesis and development of human breast cancer.