Objective:To determine if growth hormone (GH) and glutamine (Gln) reduce the time and dose of parenteral nutrition (PN) in patients with short bowel syndrome. Methods: Sixteen PN-dependent patients with short bowel syndrome were randomly assigned to Group A (control, oral Gln [0.6 g/kg/d] + GH placebo + diet, n= 5), B (Gln placebo + GH [8 IU/d] + diet, n= 5), and C (Gln + GH + diet, n= 6). PN volume and content were determined by standard criteria based on clinical measurements. After 4 weeks, GH and GH placebo were discontinued, but the diet with Gln or Gln placebo was continued for 12 weeks. Results: Group B showed greater reductions in PN volume, PN calorie and PN frequency than Group A (P

Objective:To explore the clinical characteristics and treatment of a family with inherited generalized epilepsy with febrile seizures plus (GEFS + ) caused by the KCNT2 gene mutation and review the literature. Methods:Clinical data of a child with GEFS + and his family members who visited Department of Pediatric Neurology, Guangzhou Women and Children′s Medical Center in May 2019 were collected.DNA samples were collected from the peripheral blood of the proband, his parents, his elder brother, and his maternal grandparents, and genetic analysis and verification were performed using the next-generation sequencing technique.Using " KCNT2" as the key word, literature was retrieved from PubMed, China National Knowledge Infrastructure and Wanfang databases (up to August 2019). Results:The proband was a 3-year-old boy who was admitted to Guangzhou Women and Children′s Medical Center because of frequent epileptic seizures in the past 5 months.He presented with a binocular gaze and experienced 3 to 8 times of extremities myoclonic-spastic epileptic attacks every day.He had a history of 3 times of febrile seizures at the age of 2 years old.His seizures were refractory to Sodium valproate, Topiramate, Nitrazepam and Levetiracetam.His elder brother and mother had a history of childhood febrile seizures.Other members in the family had no history of convulsion.Ictal electroencephalogram showed general 1 Hz high voltage spike-slow waves.A heterozygous nonsense mutation of KCNT2 gene c. 574C>T(p.Q192X) that was never reported previously was detected in the proband, his brother, mother and maternal grandmother.Furthermore, no other family members carried the mutation at the c. 574 locus of the KCNT2 gene.No article in Chinese was found, and 2 articles in a language other than Chinese provided the complete data of 3 sporadic cases.Together with 4 cases in the family studied in this article, there were 7 cases and 4 mutation sites in KCNT2 gene.Of these mutations, there were 3 missense mutations and 1 nonsense mutation.Three sporadic patients presented with early infantile epileptic encephalopathy.The family of this study was characterized with febrile seizures and febrile seizures plus. Conclusions:A de novo mutation and phenotype of the KCNT2 gene is found in a family with GEFS + .It would expand the gene mutation spectrum and provide basis for family genetic counseling. KCNT2 mutation induced GEFS + is refractory to antiepileptic drugs.

Kv4.3 channel is present in many mammalian tissues, predominantly in the heart and central nervous system. Its currents are transient, characterized by rapid activation and inactivation. In the hearts of most mammals, it is responsible for repolarization of the action potential of ventricular myocytes and is important in the regulation of the heart rate. Because of its central role in this important physiological process, Kv4.3 channel is a promising target for anti-arrhythmic drug development. Jingzhaotoxin-V (JZTX-V) is a novel peptide neurotoxin isolated from the venom of the spider Chilobrachys jingzhao. Whole-cell patch clamp recording showed that it partly blocked the transient outward potassium channels in dorsal root ganglion neurons of adult rats with an IC(50) value of 52.3 nmol/L. To investigate the effect of JZTX-V on Kv4.3 channel, JZTX-V was synthesized using the solid-phase chemical synthesis and separated by reverse phase high performance liquid chromatography (HPLC). The purity was tested by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MOLDI-TOF mass spectrometry). Two-electrode voltage-clamp technique was used to characterize the action of JZTX-V on Kv4.3 channels expressed in Xenopus laevis oocytes. As a result, JZTX-V displayed fast kinetics of inhibition and recovery from inactivation. Furthermore, it could inhibit Kv4.3 channel current in a time- and concentration-dependent manner with an IC(50) value of 425.1 nmol/L. The application of JZTX-V affected the activation and inactivation characteristics of Kv4.3 channel and caused a shift of the current-voltage relationship curve and the steady-state inactivation curve to depolarizing direction by approximately 29 mV and 10 mV, respectively. So we deduced that JZTX-V is a gating modifier toxin of Kv4.3 channel. Present findings should be helpful to develop JZTX-V into a molecular probe and drug candidate targeting to Kv4.3 channel in the myocardium.
Animals ; Ganglia, Spinal ; cytology ; Neurons ; drug effects ; Neurotoxins ; pharmacology ; Oocytes ; Patch-Clamp Techniques ; Peptides ; pharmacology ; Potassium Channel Blockers ; pharmacology ; Rats ; Shal Potassium Channels ; metabolism ; Spider Venoms ; pharmacology ; Xenopus laevis

OBJECTIVE: To investigate the application value of fusion indocyanine green fluorescence imaging(FIGFI)in the surgical treatment of recurrent hepatocellular carcinoma.METHODS: From January 2015 to January 2018,theclinical data of 12 patients with recurrent hepatocellular carcinoma who received surgical treatment guided by three-dimensional visualization technology combined with FIGFI at Department of Hepatobiliary Surgery,Zhujiang Hospital,Southern Medical University were analyzed. High-quality thin-slice CT data were collected for three-dimensionalvisualization and reconstruction,individualized liver segmentation and preoperative surgical planning. FIGFI was used todefine the demarcation of tumors,to determine the extent of hepatectomy,and to detect small hepatocellular carcinomaand metastases during the operation. The use of FIGFI in combination with three-dimensional visualization imagesguided the precision hepatectomy. postoperative efficacy was observed by follow-up.RESULTS: All the 12 patientsunderwent successful three-dimensional reconstruction and liver resection. Preoperative CT revealed 12 lesions(80%);intraoperative FIGFI detected 15 lesions(100%);of the three newly discovered lesions,2 were hepatocellular carcinomaand 1 was cirrhosis nodule. The operation time was 267.5(183,345)minutes and the amount of bleeding was 287.5(62.5,513.5)m L. No severe complications such as abdominal hemorrhage,abdominal infection,bile leakage and liverfailure occurred postoperatively. No death occurred during the perioperative period. The median follow-up period was 29 (3-36)months; during the follow-up period, 1 casewas lost and 4 cases had metastasis or recurrence.All the patients were alive at the end of the follow-up.CONCLUSION: FIGFI is helpful to achieveanatomical, functional and radical hepatectomy inthe treatment of recurrent hepatocellular carcinoma.

Objective:To explore the clinical characteristics of interleukin-6 (IL-6) and interleukin-8 (IL-8) in cerebrospinal fluid (CSF) of children with bacterial meningitis (BM) and provide reference for clinical diagnosis and treatment of BM.Methods:The clinical data of BM children hospitalized in Women and Children′s Medical Center Affiliated to Guangzhou Medical University from December 2019 to March 2022 were collected and retrospectively analyzed in this case series study.Cytokines in CSF of these children were detected at least twice during the treatment. t test, Mann-Whitney test or analysis of variance were carried out for statistical analysis. Results:There were 40 patients included in this study.The age of onset was 2(1, 8) months, ranging from 2 days to 8 years, and the length of time from onset to hospitalization was (15±17) days, ranging from 1 day to 69 days.The main symptoms at the onset were fever (40 cases, 100%), poor mental state (16 cases, 35.0%), convulsion (9 cases, 22.5%), and vomiting (9 cases, 22.5%).According to pathogens, the patients were divided into the Streptococcus agalactia group (GBS group, 9 cases), Streptococcus pneumoniae group (SP group, 9 cases), other bacteria group (9 cases), and unknown bacteria group (13 cases).The levels of cytokines in the CSF of BM children were increased, along with significantly elevated levels of IL-6 and IL-8 within 1 st week of BM, followed by the peak at 2 nd-3 rd weeks, and then levels of IL-6 and IL-8 presented an overall decreasing trend with the progression of BM.The level of IL-6 in CSF of 10 cases significantly decreased in the 4 th week of BM [within 2 weeks: 773.5(164.1, 1 781.2) ng/L vs. 4 th week: 10.8(2.2, 21.1) ng/L, P=0.005].Such statistical differences didn′t occur to the level of IL-8 [within 2 weeks 182.9(33.6, 657.7) ng/L vs. 4 th week: 92.9(22.6, 226.6) ng/L, P=0.303].After effective antibiotic therapy, 6 patients had elevated white blood cell count in CSF during the 4 th-20 th weeks, with or without repeating intermittent fever.Among them, 4 cases of GBS and 1 case of SP were negative for pathogens in CSF during the retest after treatment, and the levels of IL-6 and IL-8 [(149.1-4 218.6) ng/L and (124.2-1 890.3) ng/L, respectively] in CSF were elevated.Low-dose glucocorticoid was administered for anti-inflammatory treatment, with additional gamma globulin for 1 case and Ibuprofen instead for 1 case.Subsequently, the fever completely subsided.The white blood cell count in CSF decreased significantly ( P=0.024). Conclusions:The levels of IL-6 and IL-8 in CSF increase significantly in the acute phase of BM and generally decrease with the progression of BM.If they are still significantly elevated in the later course of BM, it should be noted that an intracranial hyperinflammatory response may occur, especially when the pathogenic bacteria are GBS or SP.

Background and purpose:Leptomeningeal metastasis is a form of central nervous system metastasis of melanoma.High mobility group A2(HMGA2)has been proven to play an important role in the occurrence and development of various tumors,but its biological functions in leptomeningeal metastatic melanoma cells remain unclear.On the basis of building mouse models of central nervous system metastasis of melanoma,this study investigated the differences in cell migration and cell proliferation among leptomeningeal metastatic melanoma cells,primary site melanoma cells and brain parenchymal metastatic melanoma cells,and further clarified the effects of differentially expressed gene HMGA2 on cell migration and proliferation of leptomeningeal metastatic melanoma cells.Methods:B16 mouse melanoma cells(B16-parental cells,B16-Par)stably expressing tdTomato and luciferase were generated by lentiviral infection.Subsequently,B16 specific brain parenchymal metastatic cells(B16-brain metastatic cells,B16-BrM)and B16 specific leptomeningeal metastatic cells(B16-leptomeningeal metastatic cells,B16-LM)were collected after adaptive screening of metastatic sites in vivo.The differences in migration and proliferation among B16-Par,B16-BrM and B16-LM were assessed by wound healing assay and cell counting kit-8(CCK-8).RNA sequencing(RNA-seq)was used to analyze differential gene expression in B16-Par,B16-BrM and B16-LM,and HMGA2 gene specifically upregulated in B16-LM was screened out.The results were verified by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot.Gene ontology(GO)analysis was performed for genes which were upregulated in B16-LM specifically.siRNA was used to interfere with the expression of HMGA2 gene in B16-LM,and the knock-down effect was verified by RTFQ-PCR and Western blot.The effects of knocking down HMGA2 on cell migration and proliferation were detected by wound healing assay and CCK-8 assay.Using GSE174401 data in Gene Expression Omnibus(GEO),the specificity of HMGA2 gene expression in leptomeningeal metastatic melanoma cells from patients was verified.Results:Compared with Par cells,tumor cells screened by the brain environment were more likely to colonize the central nervous system.B16-LM had stronger migration and proliferation abilities,and upregulated the expression of HMGA2 gene.GO analysis revealed that HMGA2 was associated with many biological processes such as angiogenesis and cell proliferation.When the expression of HMGA2 gene was knocked down,the migration and proliferation of B16-LM could be inhibited.HMGA2 was upregulated in leptomeningeal metastatic melanoma cells from patients.Conclusion:Leptomeningeal metastatic melanoma cells had relatively unique cellular characteristics,which promoted cell migration and proliferation by upregulating HMGA2 gene expression.

OBJECTIVES: This study aims to investigate the effect of RhoE expression on the migration and invasion of tongue squamous cell carcinoma (TSCC). METHODS: Forty-eight TSCC cases were selected from the Maxillofacial Surgery Center of Qingdao Municipal Hospital from 2017 to 2019. The expression of RhoE in the specimens (TSCC and adjacent tissues) was detected by immunohistochemistry, and RhoE mRNA and protein were extracted to further detect the expression of RhoE. SCC-4 and CAL-27 cells were selected for RESULTS: The expression level of RhoE in TSCC was significantly lower than that in adjacent tissues ( CONCLUSIONS RhoE expression is low in TSCC. Over expression RhoE in TSCC can significantly decrease its migration and invasion abilities. Hence, RhoE may play an important role in regulating the metastasis and invasion of TSCC and provide a new target for gene therapy.
Carcinoma, Squamous Cell ; Cell Line, Tumor ; Humans ; Matrix Metalloproteinase 2 ; Neoplasm Invasiveness ; Tongue ; Tongue Neoplasms ; rho GTP-Binding Proteins/genetics* ; rho-Associated Kinases

OBJECTIVES: This study aimed to explore the effects of silencing isoprenylcysteine carboxyl methyltransfe-rase (Icmt) through small interfering RNA (siRNA) interference on the proliferation and apoptosis of tongue squamous cell carcinoma (TSCC). METHODS: Three siRNA were designed and constructed for the Icmt gene sequence and were then transfected into TSCC cells CAL-27 and SCC-4 to silence Icmt expression. The tested cells were divided as follows: RNA interference groups Icmt-siRNA-1, Icmt-siRNA-2, and Icmt-siRNA-3, negative control group, and blank control group. The transfection efficiency of siRNA was detected by the fluorescent group Cy3-labeled siRNA, and the expression of Icmt mRNA was screened by quantitive real-time polymerase chain reaction (qRT-PCR) selected the experimental group for subsequent experiments. The expression of Icmt, RhoA, Cyclin D1, p21, extracellular regulated protein kinases (ERK), and phospho-extracellular regulated protein kinases (p-ERK) were analyzed by Western blot. The proliferation abilities of TSCC cells were determined by cell counting kit-8 assay. The change in apoptosis was detected by AnnexinV-APC/propidium staining (PI) assay. Cell-cycle analysis was conducted by flow cytometry. RESULTS: The expression of Icmt mRNA and protein in TSCC cells significantly decreased after Icmt-siRNA transfection ( CONCLUSIONS Silencing Icmt can effectively downregulate its expression in TSCC cells, reduce the RhoA membrane targeting localization and cell proliferation, and induce apoptosis. Thus, Icmt may be a potential gene therapy target for TSCC.
Apoptosis ; Carcinoma, Squamous Cell ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Protein Methyltransferases ; RNA, Small Interfering ; Tongue ; Tongue Neoplasms

OBJECTIVES: The effect of isoprenylcysteine carboxymethyltransferase (ICMT) silencing on the migration and invasion of tongue squamous cell carcinoma was investigated by constructing the small interfering RNA (siRNA) of ICMT. METHODS: Through liposomal transfection, siRNA was transfected into human tongue squamous cell carcinoma CAL-27 and SCC-4 cells (ICMT-siRNA group) with a negative control group (transfected with NC-siRNA) and a blank control group (transfected with a transfection reagent but not with siRNA). Quantitative real-time polymerase chain reaction was performed to analyze the mRNA expression of ICMT and RhoA in each group of cells after transfection and to measure the silencing efficiency. Western blot was applied to examine the expression levels of ICMT, total RhoA, membrane RhoA, ROCK1, matrix metalloproteinase (MMP)-2, and MMP-9 proteins in each group. The migration and invasion abilities were evaluated via wound healing and Transwell motility assays. RESULTS: After CAL-27 and SCC-4 cells were transfected with ICMT-siRNA, the expression levels of ICMT genes and proteins decreased significantly in the experimental group compared with those in the negative and blank control groups ( CONCLUSIONS The migration and invasion abilities of CAL-27 and SCC-4 cells were reduced significantly after the transfection of ICMT-siRNA, and the involved mechanism might be related to the RhoA-ROCK signaling pathway.
Carcinoma, Squamous Cell ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Humans ; Neoplasm Invasiveness ; Protein Methyltransferases ; RNA, Small Interfering ; Tongue ; Tongue Neoplasms ; Transfection ; rho-Associated Kinases

Objective: To retrospectively analyze the data of Chinese patients with newly diagnosed acute promyelocytic leukemia (APL) to preliminarily discuss the clinical and cytogenetic characteristics. Methods: From February 2004 to June 2020, patients with newly diagnosed APL aged ≥ 15 years who were admitted to the Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Science & Peking Union Medical College were chosen. Clinical and laboratory features were retrospectively analyzed. Results: A total of 790 cases were included, with a male to female ratio of 1.22. The median age of the patients was 41 (15-76) years. Patients aged between 20 and 59 predominated, with 632 patients (80%) of 790 patients classified as low and intermediate risk and 158 patients (20%) of 790 patients classified as high risk. The white blood cell, platelet, and hemoglobin levels at diagnosis were 2.3 (0.1-176.1) ×10(9)/L, 29.5 (2.0-1220.8) ×10(9)/L, and 89 (15-169) g/L, respectively, and 4.8% of patients were complicated with psoriasis. The long-form type of PML-RARα was most commonly seen in APL, accounting for 58%. Both APTT extension (10.3%) and creatinine>14 mg/L (1%) are rarely seen in patients at diagnosis. Cytogenetics was performed in 715 patients with newly diagnosed APL. t (15;17) with additional chromosomal abnormalities were found in 155 patients, accounting for 21.7%; among which, +8 was most frequently seen. A complex karyotype was found in 64 (9.0%) patients. Next-generation sequencing was performed in 178 patients, and 113 mutated genes were discovered; 75 genes had an incidence rate>1%. FLT3 was the most frequently seen, which accounted for 44.9%, and 20.8% of the 178 patients present with FLT3-ITD. Conclusions: Patients aged 20-59 years are the most common group with newly diagnosed APL. No obvious difference was found in the ratio of males to females. In terms of risk stratification, patients divided into low and intermediate risk predominate. t (15;17) with additional chromosomal abnormalities accounted for 21% of 715 patients, in which +8 was most commonly seen. The long-form subtype was most frequently seen in PML-RARα-positive patients, and FLT3 was most commonly seen in the mutation spectrum of APL.
Adult ; Aged ; Chromosome Aberrations ; Cytogenetics ; Female ; Humans ; Leukemia, Promyelocytic, Acute/genetics* ; Male ; Middle Aged ; Mutation ; Oncogene Proteins, Fusion/genetics* ; Retrospective Studies ; Young Adult