Objective To investigate the changes in serum hyaluronic acid level at different periods of experimental piggyback liver transplantation and its significance. Methods Fifteen pairs of healthy pigs of both sexes weighing (28.3 + 5.0)kg undergoing liver transplantation were studied. The donor pigs were slightly smaller than the recipient pigs. The recipient pigs were premedicated with intramuscular ketamine 8mg kg-1 and atropine 0.02mg-kg-1 .Anesthesia was induced with propofol 2mg-kg-1 , fentanyl 0.002mg-kg-1 and vecuronium 0.1mg-kg-1 iv. After tracheal intubation the animals were mechanically ventilated. CVP line was placed via internal jugular vein. Carotid artery was cannulated for continuous BP monitoring. Anesthesia was maintained with iv propofol, fentanyl and vecuronium. Circulatory stability was maintained by infusion of crystalloid, colloid, plasma and whole blood of pig. Blood samples were taken from peripheral vein before operation (T0), pre-anhepatic phase (T1), anhepatic phase (T2) and neohepatic phase (T3) for determination of serum hyaluronic acid concentration by radioimmunoassay. At the same time liver function tests, ALT, AST, y-GT were also examined. Results The average time of liver transplantation was (343+74) min. SBP decreased by (23.51+5.87 ) mm Hg and DBF by (11.35+7.81) mm Hg after induction of anesthesia. Serurn hyaluronic acid level was 267ug-L-1 before operation (T0) and rose to 1743ug-L-1 at T1 , and 9530ug-L-1 at T2 (P

OBJECTIVE: To investigate the expression and clinicopathological significance of the cell cycle regulators cyclin E, cyclin D1, p21, p16 in laryngeal carcinogenesis tissus. METHOD: The expression of cell cycle regulators were detected by flow cytometry method in 23 cases of polyps of vocal cord, 69 cases of laryngeal precancerous change and 33 cases of laryngeal squamous cell carcinoma (LSCC), which tissue was paraffin embedded, sliced, dewaxed, and prepared into the cell suspension, then fluorescently labeled by cyclin E, cyclin D1, p21 and p16. RESULT: In polyps of vocal cord, laryngeal precancerous change and LSCC, The positive expression rate of cyclin E and cyclin D1 were respectively 13.04%, 20.29D, 42.420 and 26.09%, 43.48% and 93.94%. The positive expression rate of p16 and p21 were respectively 61.90%, 40.98%, 14.28% and 47.62%, 23.81%, 26.23%. Those showed the positive expression rate of cyclin D1, cyclin E gradually decreased from vocal cord polyps, laryngeal precancerous change to LSCC, (P < 0.05, P < 0.01), while the positive expression rate of p21 and p16 gradually decreased (P < 0.01). CONCLUSION The abnormal expression of cell cycle regulatory factors is the molecular events of laryngeal carcinoma. High expression of positive regulatory factors cyclin D1 and cyclin E, and low expression of negative regulatory factors p16 and p21, which showed the imbalance of multiple positive and negative regulatory factors related with cell cycle play an important role in the occurrence of laryngeal cancer.
Adult ; Cyclin D1 ; metabolism ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Female ; Flow Cytometry ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Oncogene Proteins ; metabolism

Objective To analyze histopathological and clinical features of dermatofibroma,and to explore the relationship between them.Methods Clinical and histopathological data were collected from 150 patients with histopathologically confirmed dermatofibroma in Department of Pathology,Shanghai Skin Disease Hospital from September 2017 to August 2018,and analyzed retrospectively.Results Among the 150 patients,65 were males,and 85 were females.Their age was 42 ± 13.8 years,and the course of disease ranged from 3 months to 30 years.Some of the patients had concomitant symptoms,mainly manifesting as itching,some had spontaneous pain and mild tenderness,and 18 patients had a history of injury,insect bite or infection at lesion sites.Skin lesions mainly occurred on the extremities (107 cases,71.3％),and most were solitary (105 cases,70％).Before pathological examinations,102 cases were clinically diagnosed as dermatofibroma,16 as epidermoid cyst,13 as pigmented nevus,3 as keloid,12 as skin mass,1 as malignant melanoma,1 as xanthogranuloma,1 as prurigo nodularis,and 1 as neurofibroma.Among 169 hematoxylin and eosin (HE)-stained sections,25 (14.8％) appeared to be consistent with aneurysmal dermatofibroma,66 (39.1％)with cellular dermatofibroma,36 (21.3％) with sclerosing dermatofibroma,and 22 (13.0％)with epithelioid dermatofibroma.Coexistence of two or more subtypes could be seen in 12 sections.There were also a few new variants,such as dermatofibroma with hyperplastic sweat duct (1 case),deep dermatofibroma (3 cases),dermatofibroma with epithelioid cells intermingled with hyperplastic collagen (1 case).The duration of aneurysmal dermatofibroma varied from 7 months to 30 years,and most manifested as skin masses on the lower extremities.A relatively short course of disease was observed in patients with cellular dermatofibroma,who often visited a hospital several months after the onset,and cellular dermatofibroma was commonly observed on the extremities and frequently accompanied with itching and pain.The duration of sclerosing or atrophic dermatofibroma was usually long for years or decades,and it commonly occurred on the upper limbs without concomitant symptoms.Epithelioid dermatofibroma of varied durations had various clinical manifestations,frequently occurred on the lower limbs without concomitant symptoms.Conclusions The clinical and pathological manifestations of dermatofibroma are diverse.Different dermatofibroma lesions can share similar typical histopathological manifestations,and atypical pathological features can interfere with the diagnosis of dermatofibroma.

OBJECTIVE: To explore the genetic basis for a patient diagnosed with tuberous sclerosis complex (TSC). METHODS: Peripheral blood samples of the patient and his parents were collected for the extraction of genomic DNA. Next generation sequencing (NGS) was carried out to detect potential variant, and the result was verified by Sanger sequencing. RESULTS: The patient was found to harbor a heterozygous c.1053delG (p.Glu352SerfsX10) frameshifting variant of the TSC2 gene. The same variant was not found in his unaffected parents and 100 unrelated healthy controls. Based on the American College of Medical Genetics and Genomics guidelines, the variant was predicted to be pathogenic (PVS1+PS2+PM2). CONCLUSION The novel c.1053delG (p.Glu352SerfsX10) frameshifting variant of the TSC2 gene probably underlay the TSC in this patient.
Genomics ; Heterozygote ; Humans ; Mutation ; Tuberous Sclerosis/genetics* ; Tuberous Sclerosis Complex 2 Protein/genetics*

Objective:To analyze clinical phenotypes and pathogenic mutations of a patient with classic tuberous sclerosis complex.Methods:Clinical data was collected from a patient with classic tuberous sclerosis complex. Next-generation sequencing was performed to screen pathogenic gene variants, and Sanger sequencing to verify the mutations. Minigene plasmids were constructed and transfected into the human renal epithelial cell line 293T, and RNA was extracted for transcriptional analysis.Results:The patient clinically presented with recurrent epileptic seizures, facial angiofibroma, periungual fibroma, pulmonary lymphangioleiomyomatosis, renal angiomyolipoma and multiple osteosclerosis. Next-generation sequencing revealed a suspected pathogenic variant in the TSC2 gene in the patient. Sanger sequencing identified a heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene in the patient, but not in his parents or 100 unrelated healthy controls. Moreover, this mutation had not been previously reported. The minigene experiment showed changed mRNA sequence of the TSC2 gene in this patient with loss of the authentic splice site in exon 4 and insertion of a 74-bp intron, which shifted the splice site 90 bp downstream (r.336delins336+16_336+90) .Conclusion:The novel heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene can lead to aberrant splicing, and may contribute to tuberous sclerosis complex in this patient.

OBJECTIVETo detect the expression of SCUBE3 in human osteosarcoma cell lines and surgical specimens of osteosarcomas and investigate its association with the patients' prognosis.

METHODSThe expression of SCUBE3 protein was detected in 5 osteosarcoma cell lines using Western blotting. CCK8 assay was used to assess the effect of SCUBE3 suppression mediated by two specific small interfering RNAs (siRNAs) on the proliferation of U2OS osteosarcoma cells, and the cell cycle changes were detected using flow cytometry. Immunohistochemistry was performed to detect the expression of SCUBE3 in 60 osteosarcoma tissues, and Kaplan-Meier method was performed for survival analysis of the patients.

RESULTSCompared with osteoblast hFOB1.19 cells, the osteosarcoma cell lines all showed significantly higher expressions of SCUBE3. In U2OS cells, suppression of SCUBE3 expression by siRNA significantly inhibited the cell proliferation (P<0.05). Kaplan-Meier survival analysis indicated that patients with high SCUBE3 expression had worse prognosis than those with low SCUBE3 expression (P=0.036).

CONCLUSIONSCUBE3 is up-regulated in the 5 osteosarcoma cell lines and in primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high SCUBE3 expression in osteosarcoma tissues is associated with a poor prognosis of the patients, suggesting that SCUBE3 can serve as a potential therapeutic target for osteosarcoma.

Bone Neoplasms ; metabolism ; pathology ; Calcium-Binding Proteins ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Humans ; Immunohistochemistry ; Kaplan-Meier Estimate ; Osteosarcoma ; metabolism ; pathology ; Prognosis ; RNA, Small Interfering ; Up-Regulation

OBJECTIVETo detect the expression of CXCL14 in human osteosarcoma cell lines and tissues and investigate its association with the prognosis of the patients.

METHODSRT-PCR, enzyme-linked immunosorbent assay (ELISA) and real-time PCR were used to detect the expression of CXCL14 in 4 osteosarcoma cell lines and in 40 pairs of osteosarcoma tissues and adjacent muscular tissues. CCK8 assay and colony formation assay was used to assess the effect of CXCL14 suppression mediated by two specific siRNAs on the proliferation of U2OS osteosarcoma cells. Immunohistochemistry was performed to detect the expression of CXCL14 in 58 osteosarcoma tissues, and Kaplan-Meier method and log-rank test were performed for survival analysis of the patients.

RESULTSSignificant up-regulation of CXCL14 expression was found in the osteosarcoma cell lines and in osteosarcoma tissues compared with the adjacent muscles (P<0.01). In U2OS cell, suppression of CXCL14 expression by siRNA significantly inhibited the cell proliferation (P<0.01) and colony formation rate (P<0.05). Kaplan-Meier survival analysis indicated that patients with high CXCL14 expression had worse prognosis than those with low CXCL14 expression (P=0.02).

CONCLUSIONCXCL14 is up-regulated in both osteosarcoma cell lines and primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high CXCL14 expression in osteosarcoma tissues is associated with a poor prognosis, suggesting the that CXCL14 serve as a potential therapeutic target for osteosarcoma.

Bone Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Chemokines, CXC ; metabolism ; Humans ; Osteosarcoma ; metabolism ; pathology ; Prognosis

Objective:To evaluate the safety and efficacy of the technique of minimally invasive separation surgery with small incision and free hand screw placement in patients with spinal metastases.Methods:Retrospectively reviewed the clinical data of 49 consecutive patients from May 2019 to December 2019 who underwent minimally invasive separation surgery with small incision and free hand screw placement for metastatic spinal tumors. Among them, there were 21 males with an average age of 55.62±2.97 years (range: 26-75 years) and 28 females with an average age of 52.50±1.76 years (range: 34-72 years). For patients who have primary tumor history with multiple metastases, routine pre-operative biopsy is not required; but for patients whose primary tumor is unknown and who have no history of tumor, pre-operative biopsy diagnosis is required. Before operation, Karnofsky Performance status (KPS) scoring system was used to evaluate the general condition of patients, Spinal Instability Neoplastic Score (SINS) scoring system was used to evaluate the spine stability, epidural spinal cord compression (ESCC) grading system was performed to access the degree of spinal cord nerve compression, and Frankel grading system was used to evaluate the neurological function. For patients who meet inclusion and exclusion criteria ware performed for decompression and internal fixation by a minimally invasive separation surgery with small incision and free hand screw placement. The demographic, neurological function, complications and perioperative data were collected and analyzed, including pre-operation neurological function, operation time, intraoperative blood loss, postoperative suction drainage, drainage tube extraction time, complications rates, hospital stay, and assessment of neurological recovery at 4 weeks after surgery.Results:Preoperative coil embolization was performed in 1 patient with kidney cancer. The mean intraoperative blood loss was 748.60±79.39 ml. Comparison of intraoperative blood loss of 12 rich blood supply (liver cancer, kidney cancer, thyroid cancer) and 37 poor blood supply spine metastases (970.80 ml vs 676.50 ml) was not statistically significant ( P>0.05). The average operation time was 213.40±9.87 min. The operation involved 1 segment was performed in 41 patients (83.67%) and 8 patients had separation of 2 or more segments. The post-operative drainage before discharge was 494.02±63.30 ml. The average drainage tube retention time was 4.50±0.26 d and the average length of hospital stay was 7.35±0.38 d. The post-operative hospitalization was 5.31±0.29 d. 79.59% of patients had the neurological functions of Frankel grade C and D before surgery and 95.92% of patients exhibited stable and improved function at 4 weeks after surgery which was significantly improved comparing with that before surgery ( P<0.05). The complications occurred were dural rupture (1 case), infection (1 case) and hematoma (1 case). Comparison:The minimally invasive separation surgery with small incision and free hand screw placement could achieve less trauma, low complications rate, rapid postoperative recovery. It is also comparable to the traditional open separation surgery in terms of spinal stability and improvement of neurological functions. It is an excellent alternative for patient with spinal metastases.