[Multiple pulmonary nodules with cavity formation].

A 64-year-old female patient was admitted to Beijing Chao-Yang Hospital on February 21, 2023 because of right-sided chest pain for more than 4 years and left-sided chest pain for more than 9 months. She had a past medical history of previous tuberculosis and rheumatoid arthritis. A chest CT in October 2018 revealed multiple pulmonary nodules. A CT-guided biopsy showed no tumors, and adenosine deaminase levels in the pleural effusion were elevated, suggesting a high likelihood of tuberculosis. As a result, anti-tuberculosis treatment was initiated in March 2019. In December 2019, she underwent a right lower lobe resection due to localized hydropneumothorax on the right side. Postoperative pathology unveiled granulomatous inflammation with necrosis. A chest CT in May 2020 showed a significant increase in nodules and cavities. In January 2023, a diagnosis of cryptococcal pneumonia was considered, and she was prescribed oral fluconazole. Finally, the diagnosis of pulmonary rheumatoid nodules was confirmed after a pathological consultation of the postoperative specimen. After one month of treatment with oral prednisone and mycophenolate mofetil, a follow-up chest CT showed improvement. It was recommended that she continue with her current treatment and undergo regular chest CT scans.

[The 497th case: fever, anterior hypopituitarism, lymphadenopathy].

A 57-year-old woman was admitted to hospital with fever. She still had fever treated with multiple antibiotics, and no definite evidence for infection was found. Hypothermia and hypotension developed, and magnetic resonance imaging (MRI) examination showed enlarged anterior pituitary and multiple small nodular lesions with mild enhancement on the left side. Hormone replacement and anti-infection treatment were administrated, but fever did not improve. Remarkable lymphadenopathy was found in left supraclavicular area. The pathology of lymph node biopsy indicated peripheral T-cell lymphoma (not otherwise specified, NOS). Positron emission tomography-computed tomography (PET-CT) revealed hypermetabolism in multiple lymph nodes, infiltration of the liver and spleen. The final diagnosis were peripheral T-cell lymphoma with involvement of liver and spleen (stage Ⅳ) and anterior hypopituitarism. After chemotherapy, fever alleviated and the function of anterior pituitary recovered gradually.

[Whole transcriptome analysis and critical gene regulatory network analysis during Schistosoma japonicum infection and praziquantel treatment in mice].

OBJECTIVE: To investigate long non-coding RNA (lncRNA)-microRNA (miRNA)-messenger RNA (mRNA) interactions and identify the critical gene regulatory network during Schistosoma japonicum infections and praziquantel treatment using whole transcriptome sequencing. METHODS: A total of 110 male C57BL/6 mice were randomly divided into the control group, the infection group and the treatment group. Mice in the infection treatment and the control group were infected with S. japonicum cercariae via the abdomen, and liver specimens were sampled from 10 mice 3, 6, 8 weeks post-infection. Praziquantel treatment was given to mice in the treatment group 8 weeks post-infection, and liver specimens were sampled from 10 mice 2, 4, 6, 8, 10 weeks post-treatment. Total RNA was isolated from mouse liver specimens, and the transcriptome library was constructed for highthroughput whole transcriptome sequencing. The significant differentially expressed genes were subjected to functional annotations, Gene Ontology (GO) terms enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Correlation analysis of liver specimens was performed using R Corrplot and Himsc functions, and the lncRNAmiRNA-mRNA interaction network analysis was performed using R MixOmics and Himsc functions. RESULTS: There were 1 176 differentially expressed miRNAs, 5 270 differentially expressed mRNAs, and 2 682 differentially expressed lncRNAs between the infection group and the control group, 1 289 differentially expressed miRNAs, 7 differentially expressed mRNAs, and 69 differentially expressed lncRNAs between the treatment group and the infection group, and 1 210 differentially expressed miRNAs, 4 456 differentially expressed mRNAs, and 2 016 differentially expressed lncRNAs between the treatment group and the control group. Correlation analysis showed a higher correlation of gene expression between the treatment group and the control group. Principal component analysis showed obvious separate clustering between the infection group and the treatment group. The differentially expressed genes with significant relevance were significantly enriched in 24 GO terms, including arachidonic acid metabolic process, xenobiotic catabolic process, unsaturated fatty acid metabolic process, xenobiotic metabolic process, long-chain fatty acid metabolic process, and 8 KEGG metabolic pathways, including cholesterol metabolism, tyrosine metabolism, linoleic acid metabolism, retinol metabolism, and steroid hormone biometabolism. CONCLUSIONS: There were 23 mRNAs including Cyp2b9 and 14 lncRNAs including Rmrpr in the core position of the gene regulatory network, which may play a critical role in S. japonicum infections and praziquantel treatment, and 9 miRNAs including miR-8105 may serve as potential molecular markers for diagnosis of S. japonicum infections.

[Identification of rare variants in exons of NLRC4 gene in patients with type 1 diabetes and their impact on gene function].

Objective: To identify rare variants in exon and exon-intron boundary of containing NLR family CARD domain protein 4 (NLRC4) in type 1 diabetes (T1DM) patients, and to explore their effects on gene function. Methods: A total of 508 T1DM patients and 527 healthy controls in the Department of Metabolic Endocrinology, Second Xiangya Hospital of Central South University from August 2017 to September 2020 were selected. The case group included 264 males and 244 females, and the age [M (Q1, Q3)] was [27 (11, 43)] years. The control group included 290 males and 237 females, and their ageï¼»M（Q1，Q3）］was [47 (36, 60)] years old. Identification of rare variants in exons of NLRC4 gene in T1DM patients and healthy controls was performed and verified by next-generation sequencing and sanger sequencing. The NLRC4 gene wild-type and mutant plasmids were constructed and transfected into 293T cells. Western blot (WB) was used to detect the expression of NLRC4 protein and cleavage products of pro-cysteinyl aspartate specific proteinase(procaspase-1). Cycloheximide (CHX) was added to 293T cells transfected with wild-type or mutant NLRC4 plasmid to detect the degradation of NLRC4 protein. The localization of NLRC4 protein was detected by immunofluorescence, and the concentration of IL-1ß in the cell supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Results: The sequencing results showed that 4 patients and 2 healthy controls had a heterozygous variant c.208C>T in exon 3 of the NLRC4 gene. Two patient had a heterozygous variant c.1564T>C in exon 4, and 1 patients had c.1219G>C in exon 4. These three variants might be pathogenic variants in T1DM. In 293T cells transfected with NLRC4 wild-type and c.208C>T、c.1564T>Cc.1219G>C mutant plasmids, the expression level, degradation rate, localization of NLRC4 protein and the content of cleavage products of procaspase-1 did not change significantly. However, the concentration of IL-1ß secreted by 293T cells transfected with c.1219G>C and c.208C>T plasmid [M(Q1, Q3)] was 15.25 (12.98, 17.52) and 15.44 (13.81, 17.07) ng/L, respectively, which was lower than 18.70 (16.59, 20.81) ng/L of 293T cells transfected wild-type plasmid (P=0.020, 0.010). Conclusions: NLRC4 gene rare variants c.208C>T, c.1564T>C and c.1219G>C may not change the protein expression, degradation and localization, but c.208C>T and c.1219G>C may inhibit the secretion of IL-1ß. This result suggests that NLRC4 rare variants may have an impact on gene function.

[Prenatal pyrethroid pesticides exposure and neurodevelopment of 2-year-old children: a birth cohort study].

Objective: To investigate the influence and critical windows of prenatal exposure to pyrethroid pesticides (PYRs) on neurodevelopment of 2-year-old children. Methods: The subjects of this study were derived from the Xuanwei Birth Cohort. A total of 482 pregnant women who participated in the rural district of Xuanwei birth cohort from January 2016 to December 2018 were included. Maternal urinary concentrations of PYRs metabolites during 8-12 gestational weeks, 20-23 gestational weeks and 32-35 gestational weeks were measured with ultra high performance liquid chromatography system coupled with a tandem mass spectrometry detector. Child neurodevelopment was evaluated with the Bayley Scales of Infant and Toddler Development-Third Edition at 2 years of age. Multivariate linear regression models and binary logistic regression models were used to assess the association between PYRs exposure during pregnancy and children's neurodevelopment. Results: A total of 360 mother-child pairs had complete data on maternal urinary PYRs metabolites detection and children's neurodevelopment assessment. The detection rate of any one PYRs metabolites during the first, second and third trimester were 93.6% (337/360), 90.8% (327/360) and 94.2% (339/360), respectively. The neurodevelopmental scores of Cognitive, Language, Motor, Social-Emotional, and Adaptive Behavior of 2-year-old children were (102.3±18.9), (100.2±16.3), (102.0±20.3), (107.8±23.3) and (85.8±18.6) points, respectively. After controlling for confounding factors, 4-fluoro-3-phenoxybenzoic acid (4F3PBA, one of PYRs metabolites) exposure in the first trimester reduced Motor (ß=-5.02, 95%CI: -9.08, -0.97) and Adaptive Behavior (ß=-4.12, 95%CI:-7.92, -0.32) scores of 2-year-old children, and increased risk of developmental delay of adaptive behavior (OR=2.07, 95%CI:1.13-3.82). Conclusion: PYRs exposure during the first trimester of pregnancy may affect neurodevelopment of 2-year-old children, and the first trimester may be the critical window.

[High frequency ultrasound combined with ultrasound-guided core needle biopsy for the diagnosis of primary thyroid lymphoma].

Objective: To investigate the sonographic features of primary thyroid lymphoma (PTL) and to evaluate the clinical significance of ultrasound-guided core needle biopsy (US-CNB) in PTL. Methods: A total of 24 patients with suspected PTL in Sir Run Run Shaw Hospital from January 2013 to June 2018 were analyzed retrospectively. All cases were confirmed by pathology, of them 23 patients received US-CNB and 1 patient chose operation without US-CNB, including 5 males and 19 females, aged from 39 to 75 years old. The effectiveness and safety of 23 patients with US-CNB were evaluated, and the sonographic features of 20 patients with PTL diagnosed by pathology were analyzed. Descriptive statistical methods were used in the study. Results: In the 23 patients with suspected PTL underwent US-CNB, 18 patients were diagnosed as PTL, 4 patients were respectively diagnosed as subacute thyroiditis, anaplastic carcinoma, Hashimoto's thyroiditis, and fibro thyroiditis, and the another patient was hard to diagnose by US-CNB and then was diagnosed as PTL by surgical biopsy. The success rate of US-CNB for diagnosis of PTL was 18/19, and no severe complications occurred in the patients with US-CNB. The other case was diagnosed as PTL by surgical biopsy without US-CNB. Sonographic features of 20 cases with PTL (18 cases diagnosed by US-CNB and 2 cases by surgery or surgery biopsy) were as follows: (1) Most nodules had irregular shapes and unsmooth margins; (2) Hypoechoic or markedly hypoechoic nodules with honeycombed or cord structures were observed in most cases; (3) Calcification was rare; (4) Multiple lesions were common; (5) Abundant intralesional vascularization was commonly observed; (6) Most cases had intensification of posterior acoustic enhancement; (7) Thyroid gland enlargement or with irregular shape; and (8) PTL often accompanied with lymph nodes enlargement in lateral neck or central region. Conclusion: PTL has certain sonographic features, with assistance of US-CNB, more accurate diagnosis of PTL can be obtained.

[Artificial intelligence based on images in ophthalmology].

The application of artificial intelligence (AI) in ophthalmology will greatly reduce the workload of ophthalmologists. Machine learning is an important branch of AI, and deep learning is the most important algorithm in machine learning. At present, AI is well applied in the ophthalmic field. This article summarizes the use of AI in ophthalmology and discusses its inadequacy and future to provide reference for clinical practice. (Chin J Ophthalmol, 2021, 57: 465-469).

Relationships between diabetic nephropathy and insulin resistance, inflammation, Trx, Txnip, CysC and serum complement levels.

OBJECTIVE: To investigate the relationships between diabetic nephropathy (DN) and insulin resistance, inflammation, thioredoxin (Trx), thioredoxin-interacting protein (Txnip), Cystatin C (CysC) and serum complement levels. PATIENTS AND METHODS: A total of 119 patients with type 2 diabetes mellitus (T2DM) treated in the Endocrinology Department of our hospital from January 2017 to December 2017 were enrolled as the experiment group, while 30 healthy volunteers were selected as the control group. The expression levels of inflammatory factors, Trx, Txnip, CysC and serum complements in every subject were detected. In addition, the type 2 diabetic nephropathy rat model was established via high-fat diet and injection of low-dose streptozotocin. Blood glucose, insulin resistance indexes and 24h-urinary albumin excretion were measured, and the histomorphological characteristics of the kidney in animals were observed. RESULTS: In clinical subjects, Trx level was notably lower in the simple DM group, early DN group and clinical DN group in comparison with that in the control group. The levels of Txnip and CysC in the simple DM group, early DN group and clinical DN group were remarkably higher than those in the control group. Moreover, the expression levels of TNF-α and IL-6 in the clinical DN group were significantly elevated compared with those in the simple DM group and early DN group. In addition, C1q expression in the clinical DN group was higher than that in the simple DM group and early DN group. In model rats, HOMA-IR was distinctly higher in the DM group and DN group than that in the control group. The ratio of kidney weight to body weight (KW/BW) was evidently higher in the DN group in comparison with that in the control group and DM group. CONCLUSIONS: Insulin resistance, inflammatory factors, and levels of Trx, Txnip, CysC and serum complement C1q are related to the progression of DM.

[Clinical application of self-made drainage tubes in different layers of soft tissue for negative-pressure wound therapy in 33 patients].

From January 2014 to June 2018, 28 patients with different types of deep soft tissue injury or infection were admitted to the Affiliated Jiangyin Hospital of Medical College of Southeast University; 5 patients were admitted to the Zhengzhou First People's Hospital. There were 24 males and 9 females, aged 18-89 (40±20) years. Disposable suction tubes with holes cut on side walls were used as self-made drainage tubes. The authors placed the self-made drainage tubes on different deep soft tissue layers and wound surfaces after debridement. The effective drainage sections of the wound surface drainage tubes were wrapped with silver ion antimicrobial functional active dressings. Bio-permeable membrane was used to close the operative area. The drainage tubes in the deep layer of wound and wound surface were connected in parallel by a tee and connected to wall-hanging medical negative-pressure suction device to conduct negative-pressure wound treatment at -20.0 to -10.6 kPa. The deep drainage tubes were usually removed or changed 4 or 5 days after surgery.The drainage tubes in the wound surface were synchronously replaced when removing or replacing he drainage tubes in the deep layer of wound. On 4 to 15 days after surgery, the deep drainage tubes were removed. On 8 to 25 days after surgery, the wound surface drainage tubes were removed. Then the treatment was changed to a conventional dressing change until the wounds were completely healed or the wound bed was ready for skin grafts or tissue flaps. The indwelling time of deep drainage tubes in this group of patients was (6.2±2.8) days, and the indwelling time of wound surface drainage tubes was (12.0±3.0) days. The wound healing time was (22±5) days, the hospital stay time was (29±7) days, and wound bacteria were reduced from 6 species and 11 strains before treatment to 3 species and 4 strains after treatment. No adverse events such as wound bleeding, irritative pain, and chronic sinus occurred during treatment. Twenty-three patients were followed up for 13 to 28 months, no treatment-related complications were observed.

Effect of GGCX on the differentiation function of osteoporosis bone marrow mesenchymal stem cells through regulating TGFß/smad signaling pathway.

OBJECTIVE: Osteoporosis (OP) has a high incidence and can be found in multiple age groups. The bone marrow mesenchymal stem cells (BMSCs) have the potential for self-renewal and multi-directional differentiation, which are often used for investigating the differentiation function of osteoporosis bone marrow mesenchymal stem cells. γ-glutamyl carboxylase (GGCX) is a carboxylase-related carboxylase and was observed to be abnormally expressed in osteoarthritis. However, the role and related mechanisms of GGCX in OP have not been fully elucidated. This work aimed to evaluate the effect of GGCX on the differentiation function of BMSCs. PATIENTS AND METHODS: Sprague-Dawley rats were randomly divided into the OP group prepared by ovariectomy and sham group. GGCX expression was tested by enzyme-linked immunosorbent assay (ELISA). BMSCs were isolated from OP rats and transfected with pcDNA-GGCX plasmids. BMSC proliferation was detected by tetrazolium salt colorimetry (MTT) assay. The osteogenic and adipogenic differentiation of BMSCs was analyzed by alizarin red staining and oil red O staining. The ALP activity was determined by alkaline phosphatase (ALP) activity colorimetric assay. Real time-PCR was used to test the expressions of osteogenesis-related genes RUNX2 and OPN mRNA. Western blot was adopted to assess the TGFß/smad signaling pathway activity. RESULTS: GGCX expression was significantly decreased in the serum of OP rats compared with the sham group (p < 0.05). The transfection of pcDNA-GGCX plasmid significantly promoted BMSC cell proliferation, increased calcified nodule formation, inhibited adipogenic differentiation, enhanced ALP activity, elevated RUNX2, and OPN mRNA expressions, and upregulated TGFß1, Smad2, and Smad7 expressions (p < 0.05). CONCLUSIONS: GGCX secretion is reduced in osteoporosis. GGCX can regulate osteoporosis via promoting the TGFß/smad signaling pathway, facilitating BMSCs osteogenic differentiation, and inhibiting BMSCs adipogenic differentiation.

[Application of self-made vacuum sealing drainage device in postoperative fixation and drainage of abdominal pedicled flaps in 8 patients with deep burns of upper limbs].

From January 2013 to December 2017, 8 patients with deep burns of upper limbs were admitted to our hospital, including 6 males and 2 females, aged 23-48 years. The wound area of full-thickness burns to burns with tendon and bone injury was 4.5 cm×2.0 cm-20.0 cm×10.5 cm. After debridement, thin abdominal flaps with subdermal vascular network in the size of 5.0 cm×2.5 cm-22.0 cm×12.0 cm were applied to cover the wounds, and the donor sites were sutured directly by relaxation. The disposable suction tubes with holes cut on side walls were used as drainage tubes. The part of drainage tubes with holes were wrapped with nano-silver antimicrobial dressings and then placed at the lowest position of pedicle and donor site of abdominal flap and the space between the injured limb and the abdominal wall. The loose nano-silver antibacterial dressing was used to fill the webs of fingers and the gap between the injured limb and the abdominal wall. The transparent film dressing was used to close the surgical area and then connected with a low negative voltage electric suction device to continuously suck at a negative pressure of -15 to -10 kPa. The self-made vacuum sealing drainage device was replaced at intervals of 4 to 5 days until pedicle breakage was performed 2 to 3 weeks after operation. The pedicled abdominal flaps of 8 patients had no torsion or avulsion, no pedicle blood supply disorder, and no infection or skin erosion in the operation area, and all the flaps survived after pedicle breakage.

[Surgical outcome evaluation and prediction analysis of laparoscopic left sided hepatectomy for hepatolithiasis].

Objective: To explore the risk factors of long-term treatment outcomes and establish predicting model for laparoscopic left hepatectomy in hepatolithiasis. Methods: Clinical data of 108 patients with hepatolithiasis who underwent laparoscopic left sided hepatectomy and with complete follow-up data were retrospectively collected from June 2011 to June 2016 at the Second Affiliated Hospital of Nanchang University. Twenty-six males and 82 females were enrolled. The age was (52.4±11.7) years (range:20-80 years) , and the median follow-up time was 36 months (range: 24-83 months) . Patients were randomly divided into training group (79 cases) and validation group (29 cases) with a ratio of about 3∶1. Twenty-five preoperative and intraoperative clinical factors were selected for potential factors that might affect long-term outcomes, and quality of life was used as an surrogate evaluation index. Univariate analysis and multivariate logistic regression analysis were used to investigate the potential risk factors, and to construct and validate the predictive nomogram for surgical outcomes. Results: Among 108 patients, 10 patients (9.3%) had residual stones, 8 patients (7.4%) had recurrent stones, 12 patients (11.1%) had recurrent cholangitis and 3 patients (2.8%) died. Univariate analysis showed that history of hepatobiliary surgery, gender, activation of partial thromboplastin time, alkaline phosphatase, use of choledochoscopy, postoperative stone residual, serum creatinine, postoperative biliary drainage and operation time were risk factors that may affect long-term outcomes (all P<0.15) . Multivariate analysis showed that the history of previous hepatobiliary surgery (OR=2.305, 95% CI: 0.383-4.227, P=0.019) , postoperative biliary drainage (OR=2.043, 95% CI: 0.182-4.209, P=0.048) , operation time ≥262.5 minutes (OR=1.971, 95% CI: 0.154-4.023, P=0.045) were independent risk factor affecting long-term outcomes. Based on the above factors, the predictive nomogram model was constructed. Internal and external validations showed good discrimination (area under the curve of receiver operating curve>0.7) and calibration (Hosmer-Lemeshow test: P>0.05) performance, which indicated that the prediction effect was favorable. Conclusions: History of previous hepatobiliary surgery, postoperative biliary drainage and operation time ≥262.5 minutes are independent risk factors for long-term outcome. The predictive nomogram model based on risk factors relates to surgical outcomes presented good clinical predictive effects, which might contribute to the prediction of the long-term outcomes of laparoscopic left sided hepatectomy for hepatolithiasis.

Protective effects of melatonin on mitochondrial biogenesis and mitochondrial structure and function in the HEK293-APPswe cell model of Alzheimer's disease.

OBJECTIVE: The effects and mechanisms of melatonin on Alzheimer's disease (AD) are still not researched thoroughly. 20E2 cells (HEK293-APPswe cells) are a cellular model of AD. The modulation effects of melatonin on the structure and function of mitochondria in 20E2 cells need to be studied. MATERIALS AND METHODS: The Alzheimer's disease (AD) cell model was assessed for cell viability, expression levels of mitochondrial biogenesis factors (peroxisome proliferator-activated receptor gamma coactivator 1-alpha [PGC-1α], nuclear respiratory factor 1 [NRF1], nuclear respiratory factor 2 [NRF2], mitochondrial transcription factor A [TFAM]), mitochondrial membrane potential, Na+-K+-adenosine triphosphatase (ATPase) and cytochrome C oxidase activity, adenosine triphosphate (ATP) level, mitochondrial DNA/nuclear DNA (mtDNA/nDNA) ratio, and mitochondrial structure with and without melatonin. RESULTS: Melatonin improved 20E2 cell viability, expression of mitochondrial biogenesis factors (PGC-1α, NRF1, NRF2, TFAM), mitochondrial membrane potential, Na+-K+-ATPase, and cytochrome C oxidase activity, ATP level, mtDNA/nDNA ratio, mitochondrial structure, and decreased amyloidogenic amyloid precursor protein processing. CONCLUSIONS: Mitochondrial biogenesis disorder is associated with the pathogenesis of AD through PGC-1α-NRF-TFAM pathway, and melatonin improves the mitochondrial structure and function by enhancing mitochondrial biogenesis and decreasing amyloidogenic APP processing in Alzheimer's disease.

[Effect of TCF21 on proliferation, migration and vascular production of breast cancer cells].

Objective: To investigate the effects of transcription factor 21 (TCF21) on the proliferation, apoptosis, invasion and migration and angiogenesis of human breast cancer cells (MDA-MB-231). Methods: TCF21 overexpressing plasmid was transfected into human breast cancer cell line MDA-MB-231 by liposome transfection, and Western blot was used to detect whether the transfection was successful. The effects of TCF21 overexpression on proliferation, apoptosis, invasion and migration and angiogenesis of MDA-MB-231 were detected by MTT, DAPI, Transwell and CAM. Results: The expression of TCF21 protein in Western blot showed that the TCF21 overexpression plasmid was successfully transfected into MDA-MB-231 cells. Overexpression of TCF21 inhibited the proliferation of tumor cells and inhibit the proliferation of 48 h cells after transfection. Overexpression of TCF21 inhibited tumor invasion and migration and angiogenesis, and promoted apoptosis of tumor cells. The results of Western blot showed that the protein expression of MMP-9, VEGFA and p-VEGFR2 decreased significantly after overexpression of TCF21. Conclusions: TCF21 can inhibit the proliferation, invasion and migration, angiogenesis and apoptosis of MDA-MB-231 cells. It is suggested that TCF21 can be used as a potential site for clinical diagnosis and treatment of breast cancer.

[BRAF gene mutations in ameloblastic fibromas].

OBJECTIVE: To investigate the BRAF gene mutations in ameloblastic fibroma (AF), and to further analyze the relationship between the BRAF mutation and clinical characteristics so as to provide new reference to the study of AF's molecular pathology. METHODS: Sixteen cases diagnosed as AF at the Department of Oral Pathology, Peking University School of Stomatology between January 1990 and December 2017 were collected. Genomic DNA was extracted from formalin-fixed, paraffin embedded tissues using the QIAamp DNA Mini Kit (Qiagen, Germany) according to the manufacturer's instructions. Polymerase chain reaction (PCR) and direct sequencings were used to detect the BRAF gene mutations. The clinicopathological data, such as the age, location of the lesion, symptoms and treatments were retrospectively analyzed. RESULTS: The sixteen cases of AF involved nine women and seven men aged 2-67 years. Three lesions occurred in the maxilla and thirteen in the mandible. The most common presenting symptom of AF was a painless slowly enlarging mass with swelling. Ten patients received conservative treatment and the other six patients received radical surgery. Three cases relapsed during the study period. BRAF gene mutation was found in sixteen of all the sixteen samples analyzed (100%). The BRAF mutation was a point mutation with a thymine-adenine transversion at nucleotide 1 799 of 15 exons, resulting in a change at residue 600 that substituted glutamine for valine. This mutation was the strongest activator of the downstream RAS/RAF/MEK/ERK-MAPK signaling pathway. This helped to bring about a gain-of-function mutation due to a V600E substitution. Many studies identified that BRAF regulated survival, apoptosis, and proliferation of cells by inducing MAPK pathways activation. For the existing cases, none of the age, sex, location, recurrence and treatments had a statistically significant correlation with BRAF mutation. CONCLUSION: Our findings demonstrated high prevalence of BRAF V600E mutation in AF. The pathogenic role remains to be clarified．.

[Acquired cystic kidney disease-associated renal cell carcinoma: a clinicopathologic study of three cases].

Objective: To study the clinicopathologic, immunohistochemical (IHC), histogenetic and prognostic features of acquired cystic kidney disease-associated renal cell carcinoma (ACKD-RCC). Methods: Three cases of ACKD-RCC, including two from 401 Hospital of PLA and one from the Affiliated Hospital of Qingdao University were studied by clinical, histological and IHC analysis with review of relevant literature. Results: All the three patients were male, ranging from 46 to 78 years old. All patients had history of chronic renal failure; two patients were treated with hemodialysis for 9 years and 11 years, respectively. In two cases the tumor sizes were 2.5 cm and 3.5 cm, respectively, and the tumor border was distinct. The remaining case showed extensive renal hemorrhage with an inconspicuous mass. Microscopically, the tumor cells were arranged in cribriform, microcystic or acinar structures, with variable papillary structure in one case. Hemorrhage of varying degrees was seen in all three cases, and obvious necrosis was noted in two. The tumor cells had deeply eosinophilic cytoplasm, indistinct cell border, round or oval nuclei, and prominent nucleoli (WHO/ISUP grade 3). Mitoses were rare. Abundant oxalate crystals were seen in two cases. The renal mesenchyme of all three cases were atrophic with variable cystic changes of the renal tubules, the lining cells showed atypical hyperplasia. IHC staining showed all tumors were diffusely positive for vimentin, CD10, RCC, CAM5.2, P504s and mitochondria in the cytoplasm, and were variably positive for EMA (2/3), CK7 (1/3), CA9 (1/3) and PAX8 (3/3). All cases were negative for CD117, HMB45, Melan A and TFE3. After 3-14 months follow-up, one patient died from renal failure six months after surgery. The other two patients were alive without tumor recurrence or metastasis. Conclusions: ACKD-RCC is a very rare renal cell carcinoma. The specific cribriform structure, deeply eosinophilic cytoplasm, prominent nucleoli (WHO/ISUP grade 3), and oxalate crystals deposition, associated with the history of ACKD could aid the diagnosis. ACKD-RCC arises from the proximal renal tubule and its histogenesis might be associated with proliferation and malignant change of the atypical epithelial cells of the cystic renal tubules. ACKD-RCC may have a favorable prognosis except for tumors with sarcomatoid differentiation.

[Application value of real-time three-dimensional contrast-enhanced ultrasound for morphological assessing of hepatocellular carcinoma before radiofrequency ablation treatment].

Objective: To investigate the evaluating ability of real-time three-dimensional contrast-enhanced ultrasound (RT-3D-CEUS) for morphology of hepatocellular carcinoma (HCC) before radiofrequency ablation (RFA) treatment. Methods: Sixty patients with 88 targeted lesions were enroll in this study, who have received RFA in Hangzhou Xiasha Hospital. The pretreatment imaging data of CEUS and RT-3D-CEUS were retrospective analysis. Morphological features were assessed according to the size and shape of the lesions, and were compared between two modalities with the post-treatment response as the reference standard. Results: The size of lesion measured by CEUS and RT-3D-CEUS was similar in 70 out of 88 lesions (group A). In the other 18 lesions, the size measured by RT-3D-CEUS was larger than that by CEUS (group B). For the shape analysis, 38 lesions were spheroid by CEUS and 34 lesions by RT-3D-CEUS; 47 lesions were oval by CEUS and 41 lesions by RT-3D-CEUS; 3 lesions were irregular by CEUS and 13 lesions by RT-3D-CEUS. There were 78 lesions which shape assessed by two modalities consistently (group A'), and the other 10 lesions with inconsistent shape by two modalities (group B'). Three months follow-up results showed that 4 lesions appeared tumor residue (5.7%) in group A' and 6 lesions (33.3%) in group B'. There was significant difference between these two groups (P=0.004). In the group A', there were 6 lesions (7.7%) showed tumor residue, and 4 lesions (40.0%) in group B'. The difference between these two groups was also significant (P=0.012). Conclusion: RT-3D-CEUS could be used to assess the size and shape of HCC accurately which is associated with the response of RFA.