This paper retrospectively summarized and analyzed observation of the disease and nursing care of 17 patients with pregnancy-associated fulminant type 1 diabetes and intrauterine fetal demise.Through timely supplying blood capacity and improving renal perfusion,maintaining blood glucose homeostasis and acid-alkali,potassium balance,safety of the patients was guaranteed;by providing effective psychological nursing,puerperal dietary guidance and discharge guidance,patients' rehabilitation was improved.As a result,16 patients were in stable condition,and dead babies were delivered.Major bleeding event occurred in one patient after delivering the dead baby,and the patient developed shock as well as liver and kidney failure.The patient was transferred to ICU for further treatment and became stable and was discharged after two months.

OBJECTIVETo investigate the biological function of the hepatitis C virus (HCV)-encoded F protein in hepatocytes.

METHODSThe full-length F gene was amplified by PCR from HCV genotype 1a and cloned into plasmid pSEB-3Flag by restriction enzyme digestion and ligation. Hepatoma cell lines, Huh7 and SMMC7721, were transfected with the resultant recombinant pSEB-3Flag-F or the original pSEB-3Flag (negative control) and screened with the selective antibiotic, blasticidin. Stable F gene and protein expression was verified by RT-PCR analysis. Analysis of cell growth and cell cycle was carried out by MTS assay, crystal violet staining and flow cytometry.

RESULTSHuh7 and SMMC7721 cells transfected with pSEB-3Flag-F plasmid (Huh7-F and SMMC7721-F, respectively) uniquely expressed the F gene and protein. The Huh7-F and SMMC7721-F cells showed significantly decreased proliferation rates, compared to the respective control groups. A similar HCV F-mediated growth-inhibiting activity was observed by the cell viability assay. Furthermore, cell cycle analysis revealed that the S-phase distribution was much lower in Huh7-F (47.12%) and SMMC7721-F (30.75%) cells than in the respective controls (55.35% and 33.23%, respectively) (P less than 0.05).

CONCLUSIONStable expression of the HCV F gene reduced the in vitro proliferation rate of hepatoma cell lines, indicating that the F protein may function as a growth inhibitor of infected cells.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Hepacivirus ; genetics ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Core Proteins ; genetics ; metabolism

BACKGROUNDApoptosis is involved in the adaptive responses of bone to mechanical loading. The purpose of this study was to investigate the effects of tensile forces on osteoblast apoptosis and the related mechanism by analyzing the expression of caspases, Bcl-2, and Bax.

METHODSPrimary osteoblasts were harvested from neonatal rat calvaria and were subjected to cyclic tensile forces for 72 hours using Flexcell 4000 strain unit in Minimum Essential Medium (MEM) with 10% fetal calf serum (FCS) or with serum deprivation. Apoptosis was tested by flow cytometry using annexin V/PI staining. Caspase-3 activity was analyzed via Elisa. The gene expression of caspase-8, -9, Bcl-2, and Bax was quantified by reverse transcription (RT)-PCR.

RESULTSIn 10% FCS condition, no significant difference in cell apoptosis was found between the stretched and non-stretched osteoblast cultures. Serum withdrawal resulted in higher apoptosis rate in the osteoblasts with increased caspase-3 activity, and elevated expression of caspase-9 and Bax. Six-percent elongation of stretch attenuated the cell apoptosis induced by serum starvation, concurrent with a decrease in caspase-3 activity, a decline of caspase-8 expression, and an elevation of Bcl-2 level. On the contrary, 12% elongation of stretch increased caspase-3 activity and promoted the apoptosis with an elevated expression of caspase-8 and Bax. No significant change of caspase-9 expression was identified upon force application.

CONCLUSIONSThese results suggested that tensile forces regulate cell apoptosis of primary rat osteoblasts through caspase-3 and caspase-8 signaling cascade. Light forces rescue the cells from serum deprivation-induced apoptosis by elevating Bcl-2 expression, while heavy forces promote the apoptotic insult by inducing Bax expression.

Animals ; Apoptosis ; physiology ; Caspase 8 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Caspases ; genetics ; metabolism ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Osteoblasts ; cytology ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVEThe purpose of this article is to examine the effect of curcumin on the proliferation and metastasis of human tongue squamous cell carcinoma and analyze its mechanism.

METHODSSCC-4 were treated with curcumin of 0, 5, 10, 20, 30, 60, 100 micromol x L(-1) in 24 h. MTT assay, Matrigel invasion assay, flow cytometry and fluorescence microscopy were used to examine the effect of curcumin on the growth and metastasis of SCC-4. cDNA microarray and RT-PCR were employed to analyze the expression of genes treated by curcumin.

RESULTSThe results showed that curcumin could concentration-dependently inhibit SCC-4 cell proliferation at the concentration range from 20 to 100 micromol x L(-1). Furthermore, Matrigel invasion assay indicated that curcumin can reduce SCC-4 cell invasion under the dosage of 20, 30, 60 micromol x L(-1). Flow cytometry also showed that curcumin can influence the distribution of cell cycle of SCC-4 cell with the dosage of 20, 30, 60 micromol x L(-1). And the dosage of 30 micromol x L(-1) curcumin could lead to the recruitment of alpha-tubulin. cDNA microarray showed that 87 genes were activated and 198 genes were inhibited with the effect of curcumin. These results were validated by the real time quantitative RT-PCR.

CONCLUSIONAccording to the results, it suggests that curcumin has the potential as the leading compound for anti-cancer proliferation and invasion in oral cancer treatment, and cdc27, EGFR substrate 15, PPAR-alpha and H2A histone may play an important role among this multiple anticancer-targeting ability.

Cell Line, Tumor ; Cell Proliferation ; Curcumin ; Humans ; Mouth Neoplasms

BACKGROUNDResolvin D1 (RvD1) is a newly found anti-inflammatory bioactive compound derived from polyunsaturated fatty acids. The current study aimed to explore the protective effect of RvD1 on lipopolysaccharide (LPS)-induced acute kidney injury (AKI) and its possible mechanism.

METHODSBoth in vivo and in vitro studies were conducted. Male BALB/c mice were randomly divided into control group (saline), LPS group (LPS 5 mg/kg), RvD1 group (RvD1 5 μg/kg + LPS 5 mg/kg), and blockage group (Boc-MLP 5 μg/kg + RvD1 5 μg/kg + LPS 5 mg/kg). Boc-MLP is a RvD1 receptor blocker. The mice were intraperitoneally injected with these drugs and recorded for general condition for 48 h, while the blood and kidneys were harvested at 2, 6, 12, 24, and 48 h time points, respectively (n = 6 in each group at each time point). Human proximal tubule epithelial cells (HK-2) were randomly divided into control group (medium only), LPS group (LPS 5 μg/ml), RvD1 group (RvD1 10 ng/ml + LPS 5 μg/ml), and blockage group (Boc-MLP 10 ng/ml + RvD1 10 ng/ml + LPS 5 μg/ml). The cells were harvested for RNA at 2, 4, 6, 12, and 24 h time points, respectively (n = 6 in each group at each time point). Blood creatinine was tested by using an Abbott i-STAT portable blood gas analyzer. Tumor necrosis factor-α (TNF-α) level was detected by ELISA. Kidney pathology was observed under hematoxylin and eosin (HE) staining and transmission electron microscope (TEM). We hired immune-histological staining, Western blotting, and fluorescence quantitative polymerase chain reaction to detect the expression of RvD1 receptor ALX, nuclear factor-kappa B (NF-κB) signaling pathway as well as caspase-3. Kidney apoptosis was evaluated by TUNEL staining.

RESULTSRvD1 receptor ALX was detected on renal tubular epithelials. Kaplan-Meier analysis indicated that RvD1 improved 48 h animal survival (80%) compared with LPS group (40%) and RvD1 blockage group (60%), while RvD1 also ameliorated kidney pathological injury in HE staining and TEM scan. After LPS stimulation, the mRNA expression of toll-like receptor 4, myeloid differentiation factor 88, and TNF-α in both mice kidneys and HK-2 cells were all up-regulated, while RvD1 substantially inhibited the up-regulation of these genes. Western blotting showed that the phosphorylated-IκB/IκB ratio in LPS group was significantly higher than that in the control group, which was inhibited in the RvD1 group. RvD1 could inhibit the up-regulation of cleaved-caspase-3 protein stimulated by LPS, which was prohibited in RvD1 blockage group. RvD1 group also had a lower proportion of apoptotic nuclei in mice kidney by TUNEL staining compared with LPS group.

CONCLUSIONIn LPS-induced AKI, RvD1 could decrease TNF-α level, ameliorate kidney pathological injury, protect kidney function, and improve animal survival by down-regulating NF-κB inflammatory signal as well as inhibiting renal cell apoptosis.

Acute Kidney Injury ; chemically induced ; prevention & control ; Adaptor Proteins, Signal Transducing ; analysis ; Animals ; Apoptosis ; drug effects ; Docosahexaenoic Acids ; pharmacology ; Down-Regulation ; Kidney ; drug effects ; pathology ; Lipopolysaccharides ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; antagonists & inhibitors ; Tumor Necrosis Factor-alpha ; analysis

Objective:To summarize the clinical features and gene variations in children with Townes-Brocks syndrome (TBS).Methods:The clinical data of a female infant diagnosed with TBS caused by human spalt-like transcription factor 1 ( SALL1) gene mutation in Gansu Maternal and Child Health Hospital in May 2022 were analyzed retrospectively. Relevant articles up to July 2022 were retrieved from several databases including CNKI, VIP, Wanfang, Chinese Medical Journal Network and PubMed with the terms of " SALL1 gene" and "Townes-Brocks syndrome". Patients diagnosed with TBS caused by SALL1 gene mutation were retrieved and the clinical phenotype-genotype correlations in patients with TBS caused by frameshift mutation in SALL1 gene were analyzed and summarized. Descriptive statistical analysis was applied. Results:(1) Clinical data: The index patient was a 40-day-old girl exhibiting major clinical manifestations of polycystic kidney dysplasia, congenital external ear deformity, preaxial polydactyly and recto-perineal fistula. Whole exome sequencing and Sanger sequencing revealed a heterozygous variation of c.420delC (p.S141fs*42) in the SALL1 gene, while the same gene was found to be wild type in her parents and sister. The variant was predicted to be pathogenic (PVS1+PS2+PM2). (2) Literature review retrieved 161 cases of TBS, of which 71 were attributable to a frameshift mutation in SALL1 gene. Clinical phenotypes of the 71 cases and the index case were summarized. TBS was mainly characterized by external ear, hand and anal deformities, sometimes accompanied by hearing loss, abnormal kidney development and foot deformity. A small number of affected cases presented with rare clinical phenotypes such as abnormal eyes, hypothyroidism and abnormal development. At present, the human gene mutation database records 110 variations in the SALL1 gene, with a majority located in exon 2. The most common mutation type was frameshift variation, accounting for 52%, followed by missense variation and nonsense variation. Conclusion:TBS should be considered in children with ear, hand and anal malformations, accompanied by renal dysfunction and hearing loss, and genetic testing is recommended for timely diagnosis.

OBJECTIVE: To explore the genetic etiology of a child with Pitt-Hopkins syndrome. METHODS: A child who had presented at the Medical Genetics Center of Gansu Provincial Maternal and Child Health Care Hospital on February 24, 2021 and his parents were selected as the study subjects. Clinical data of the child was collected. Genomic DNA was extracted from peripheral blood samples of the child and his parents and subjected to trio-whole exome sequencing (trio-WES). Candidate variant was verified by Sanger sequencing. Karyotype analysis was also carried out for the child, and her mother was subjected to ultra-deep sequencing and prenatal diagnosis upon her subsequent pregnancy. RESULTS: The clinical manifestations of the proband included facial dysmorphism, Simian crease, and mental retardation. Genetic testing revealed that he has carried a heterozygous c.1762C>T (p.Arg588Cys) variant of the TCF4 gene, for which both parents had a wild-type. The variant was unreported previously and was rated as likely pathogenic based on the guidelines of the American College of Medical Genetics and Genomics (ACMG). Ultra-deep sequencing indicated that the variant has a proportion of 2.63% in the mother, suggesting the presence of low percentage mosaicism. Prenatal diagnosis of amniotic fluid sample suggested that the fetus did not carry the same variant. CONCLUSION The heterozygous c.1762C>T variant of the TCF4 gene probably underlay the disease in this child and has derived from the low percentage mosaicism in his mother.
Child ; Female ; Humans ; Male ; Pregnancy ; Intellectual Disability/genetics* ; Mosaicism ; Mothers ; Mutation ; Parents ; Transcription Factor 4/genetics*

Objective: To analyze the clinical characteristics and long-term outcomes with multicenter study for acute lymphoblastic leukemia (ALL) in children over 10 years old and adolescents. Method: Newly diagnosed ALL patients aged from 10 to 18 years old in three hospitals were included in the study from May 1(st) 2005 to April 30(th) 2015. They were received ALL-2005/2009 protocol following up to December 31(st) 2016. The clinical characteristics, outcomes and the prognostic analysis were evaluated between the two protocols. Results: Totally, 237 patients were involved in the study, 76 cases for ALL-2005 and 161 cases for ALL-2009 protocol. Complete remission (CR) after induction therapy was 94.5%. 64 (28.6%) patients relapsed with a median time of 14.5 months and 70 (29.5%) patients passed away during the following time. In long-term follow-up, the 5-year event-free survival (EFS) and 5-year overall survival (OS) of ALL patients were (63.1±3.3)% and (68.4±3.2)%. The 7-year EFS and OS were (61.0±3.5)% and (67.6±3.3)%.The 5-year EFS of intermediate risk group in ALL-2005 and ALL-2009 protocol were (73.6±6.1)% and (71.7±4.3)% with no difference (χ(2)=0.064, P=0.801). The 5-year EFS of high risk group in two protocols were (27.6±9.6)% and (33.9±9.3)%, showing no significant difference (χ(2)=0.296, P=0.586). Five years relapsed rate of two protocols were (33.8±5.7)% and (32.6±4.1)% with no difference (χ(2)=0.055, P=0.815). The mortalities were 36.8% and 29.8% separately (χ(2)=2.869, P=0.090). Univariate analysis indicated that age, male, risk, BCR/ABL translocation/t(9;22) and resistant to induction were risk prognostic factors in long-term survival (χ(2)=4.764, 4.796, 46.410, 9.560, 25.450; P=0.029, 0.029, <0.001, 0.049, <0.001). Cox multivariate analysis showed male, risk and resistant to induction were independent risk prognostic factors (RR=1.790, 2.727, 2.719; P=0.021, 0.000, 0.012). Conclusion: Protocol ALL-2009 enhanced the chemotherapy intensity in intermediate risk group with no benefit of survival. BCR-ABL fusion or t(9;22) translocation was still the risk factor of prognosis. TKI inhibitor used in these patients could improve survival. EFS rate was increased a little and death rate was decreased in ALL-2009 protocol with no significant lower relapsed rate comparing with ALL-2005 protocol.
Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; Child ; Disease-Free Survival ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Prognosis ; Remission Induction ; Risk Factors ; Translocation, Genetic

To determine heat-shock protein (Hsp)90 expression is connected with cellular apoptotic response to heat stress and its mechanism, chicken (Gallus gallus) primary myocardial cells were treated with the Hsp90 promoter, aspirin, and its inhibitor, geldanamycin (GA), before heat stress. Cellular viability, heat-stressed apoptosis and reactive oxygen species level under different treatments were measured, and the expression of key proteins of the signaling pathway related to Hsp90 and their colocalization with Hsp90 were detected. The results showed that aspirin treatment increased the expression of protein kinase B (Akt), the signal transducer and activator of transcription (STAT)-3 and p-IKKα/β and the colocalization of Akt and STAT-3 with Hsp90 during heat stress, which was accompanied by improved viability and low apoptosis. GA significantly inhibited Akt expression and p-IKKα/β level, but not STAT-3 quantity, while the colocalization of Akt and STAT-3 with Hsp90 was weakened, followed by lower cell viability and higher apoptosis. Aspirin after GA treatment partially improved the stress response and apoptosis rate of tested cells caused by the recovery of Akt expression and colocalization, rather than the level of STAT-3 (including its co-localization with Hsp90) and p-IKKα/β. Therefore, Hsp90 expression has a positive effect on cellular capacity to resist heat-stressed injury and apoptosis. Moreover, inhibition of Hsp90 before stress partially attenuated its positive effects.
Apoptosis* ; Aspirin ; Cell Survival ; Chickens* ; Heat Stress Disorders ; Heat-Shock Proteins* ; Hot Temperature* ; HSP90 Heat-Shock Proteins ; In Vitro Techniques* ; Proto-Oncogene Proteins c-akt ; Reactive Oxygen Species ; Transducers

OBJECTIVETo retrospectively analyze the clinical characteristics and the treatment outcomes of older children with acute lymphoblastic leukemia (ALL), and to evaluate the multicenter cooperation regimen (ALL-2005).

METHODSThe clinical data of 103 newly diagnosed ALL children aged 10 to 18 years old from five hospitals were enrolled in this study. They were all received ALL-2005 protocol. The clinical characteristics, the event-free survival (EFS), the overall survival (OS) and the prognostic analysis were evaluated.

RESULTS(1) Of the 103 patients, 62 were boys and 41 girls, with a median age of 12.3 years old. According to immunophenotyping, 90 (87.4% ) of 103 patients were diagnosed as B-ALL and 13 (12.6%) as T-ALL. According to risk factor, 65 (63.1%) were in intermediate risk group (MR-ALL) and 38 (36.9%) in high risk group (HR-ALL). Central nervous system leukemia (CNSL) happened in 4 (3.9%) patients at diagnosis. Of the 89 patients received chromosome test, 58 (65.2%) obtained the test results, including 21(36.2%) with aberrational chromosomes and 37 (63.8%) with normal karyotype. Of 81 patients received molecular biological test, 16 (19.8%) were positive for fusion gene. (2) After induction therapy, 97 (94.2%) obtained complete remission (CR). Twenty-eight patients relapsed with a median time of 11.9 months (ranged 2.9-57.8 months), and 38 (36.9%) patients died during the treatment. As of September 30, 2012, the median follow-up was 47 months (ranged 0.4-92.6 months). The 5-year EFS and 5-year OS of ALL patients were (60.2 ± 4.8)% and(64.1 ± 4.7)%. The 5-year EFS of MR-ALL and HR-ALL were (73.8 ± 5.5)% and (31.6 ± 8.3)% (P<0.01), the 5-year OS of MR- ALL and HR-ALL were (78.5 ± 5.1)% and (35.9 ± 8.0)% (P<0.01), respectively. (3) Cox proportion hazard regression model analysis indicated that age of 14-18 years old and BCR- ABL translocation or t(9;22) were independent risk prognostic factor for 5-year EFS.

CONCLUSIONThe incidence and prognosis in older childhood ALL were related with age, risk and biological characteristics. BCR-ABL translocation or t(9;22) was the risk factor of prognosis. ALL- 2005 protocol was recommended as the regimen for older childhood ALL.

Adolescent ; Child ; Disease-Free Survival ; Female ; Humans ; Incidence ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; therapy ; Prognosis ; Risk Factors ; Treatment Outcome