SUMMARY Tonguesquamouscellcarcinoma(TSCC)isthemostcommontypeoforalcancerandis well known for its high rate of proliferation and lymph nodal metastasis.Exploring the underlying path-ways regulating TSCC could provide novel ideas for diagnosis and prognosis of TSCC patients,as well as molecular targets for treatment of TSCC.MicroRNAs (miRNAs)are small noncoding RNAs that inhibit gene expression through the 3′untranslated regions (3′UTRs)of their target messenger RNAs.They play crucial roles in numerous biological processes,including cancer progression.Although great efforts have been made,what role miRNAs may play in the early detection and diagnosis of TSCC is not fully under-stood .Recently,our team has performed a series of basic and clinical researches in an attempt to investi-gate the relationships between miRNA expressions and prognosis of patients with TSCC and the mecha-nisms under regulation of TSCC.The results showed that miR-1 95,miR-34a,miR-29b,miR-375 and miR-26a could inhibit TSCC cells progression and development via a sophisticated network of genes.Spe-cifically,the anti-tumor effects of miR-1 95 in TSCC may be partially mediated by its inhibition of Cy-clinD1 and Bcl-2 expression.The expression of miR-34a could inhibit migration and invasion of TSCC cell lines via targeting MMP9 and MMP1 4.The function of miR-29b may be through the miR-29b/Sp1 /PTEN/AKT axis.Overexpression of miR-375 inhibited Sp1 expression by targeting the 3′untranslated re-gion of the Sp1 transcript.MEG3 and miR-26a inhibited TSCC cell proliferation,cycle progression and promoted cell apoptosis and miR-26a could increase the MEG3 expression through reduction of the ex-pression of DNMT3B in TSCC.In light of the role of those miRNAs in diagnosis and prognosis of TSCC, we reported that decreased miR-1 95 and miR-375 expression was associated with poor overall survival rate of the TSCC patients,while miR-34a expression was negatively correlated with cervical lymph node me-tastases.Furthermore,combined low expression levels of miR-26a and MEG3 emerged as an independent prognostic factor for poor clinical outcomes in TSCC patients,suggesting that combined miR-26a and MEG3 expression might prove useful as an independent biomarker of clinical prognosis among TSCC pa-tients.

Objective To analyze the clinical features of melioidosis spleen abscess.Methods A retrospective analysis of 26 cases with melioidosis spleen abscess at Hainan People′s Hospital between October 2002 and March 2015 was performed.Data with general conditions, clinical characteristics, imaging features, treatment and outcome were all extracted and analyzed.Results High fever (100.0%) and splenomegaly (73.1%) were the main clinical symptoms and signs.Laboratory examinations showed that white blood cell and neutrophil percentage increased, with a mean value of 12.6×10.9/L and 0.86, respectively.88.5% cases had diabetes mellitus which was the main underlying disease.Apart from spleen abscess, pulmonary infection (73.1%) and liver abscess (38.5%) were the majority.Imaging showed multiple small abscess (92.3%).All patients had at least one positive blood culture of Burkholderia pseudomalleiand were treated with imipenem, ceftazidime and sulfamethoxazole alone or in combination.Hospitalization days ranged between 8 to 65 days with the mean of 29.5 days.Twenty-three cases (88.5%) were cured, the remaining 3 cases (11.5%) abandoned treatment and were all lost for follow-up.Conclusions Melioidosis should be suspected in patients from epidemic areas, who have diabetes and imaging shows spleen abscess and other organ infections including pulmonary infection and liver abscess.Microbiological culture is needed immediately.

OBJECTIVETo explore the therapeutic effect of grain-sized moxibustion at "Xinshu" (BL 15) and "Shenshu" (BL 23) on early-stage Alzheimer's disease (AD) in transgenosis AD mice.

METHODSThe genotyping of amyloid precursor protein/presenilin 1(APP/PS1I) double-transgenic AD mice were detected by PCR method. Seventeen 1.5-month female transgenic (Tg 6799) mice were randomly divided into a model group (9 cases) and a treatment group (8 cases). Nine female C57BL/6J wild-type mice with identical age and background were selected into a normal group. The treatment group was treated with grain-sized moxibustion at bilateral "Xinshu" (BI. 15) and "Shenshu" (BL 23), once a day, ten treatments were considered as one course, and total 9 courses were given. The model group and normal group were treated with stimulus such as grabbing, immobilization and non-ignited moxa cone. Morris water maze (escape latency, crossing times and dwell time in the target quadrant) was applied to evaluate the learning and memory ability. Hematoxylin-Eosin (HE) staining was used to observe morphology changes in the brains of AD mice. beta-amyloid protein 1-42 (Abeta(1-42)) in the area of prefrontal cortex and hippocampus was detected by immunohistochemical method.

RESULTSAfter the treatment of grain-sized moxibustion, learning and memory ability in the treatment group was increased; compared with the model group, the escape latency was shorten, crossing times was increased, and dwell time in the target quadrant was prolonged (all P<0. 05). The crossing times and dwell time in the target quadrant in the treatment group were not significantly different from those in the normal group (both P>0.05). Compared with the normal group, the positive area and integral optical density of Abeta(1-42) in prefrontal cortex and hippocampus in the model group were increased (all P<0.01). Compared with the model group, the positive area and integral optical density of Abeta(1-42) in prefrontal cortex and hippocampus in the treatment group were reduced (P<0.01, P<0.05).

CONCLUSIONThe grain-sized moxibustion at "Xinshu" (BL 15) and "Shenshu" (BL 23) could significantly improve the learning and memory ability in APP/PS1 double- transgenic AD mice, and inhibit the over expression and accumulation of Abeta(1-42).

Alzheimer Disease ; genetics ; metabolism ; psychology ; therapy ; Amyloid beta-Peptides ; genetics ; metabolism ; Animals ; Disease Models, Animal ; Female ; Hippocampus ; metabolism ; Humans ; Learning ; Male ; Memory ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Moxibustion ; Peptide Fragments ; genetics ; metabolism ; Prefrontal Cortex ; metabolism

Background and purpose:Although bortezomib has become one of the major therapeutic agents against newly diagnosed or relapsed multiple myeloma (MM), there are some patients who become resistant to bor-tezomib and then relapse, emerging as a major obstacle to long-term survival of MM patients. It has been found that elevation of intracellular cyclic adenosine monophosphate (cAMP) levels could induce cell cycle arrest and apoptosis in MM cells,which has become an interesting approach to MM therapy. This study aimed to investigate possible effects of forskolin combined with bortezomib on bortezomib-resistant myeloma cells and further explore its mechanisms. Methods:The bortezomib-resistant MM cell lines H929-R and primary cells from patients who do not respond to bortezomib were used asin vitro models. The inlfuences of bortezomib and/or forskolin on MM cells were evaluated through cellular morphology, changes of cell distribution and apoptotic rate. Meanwhile, lfow cytometry analysis was used to detect mitochondrial transmembrane potential (ΔΨm) and the expression levels of apoptosis regulators in these cells before and after the treatment were detected by Western blot.Results:Bortezomib (20 nmol/L) synergized with forskolin (50nmol/L) to induce apoptosis of H929-R cells and bortezomib-resistant primary cells. In addition, borte-zomib synergized with forskolin to induce collapse of mitochondrial transmembrane and facilitate the degradation of anti-apoptosis proteins including Bcl-2 and Mcl-1.Conclusion:Bortezomib could synergize with forskolin to induce apoptosis in bortezomib-resistant MM cells.

Objective To investigate the drug resistance of 1 031 Mycobacterium tuberculosis to rifampicin and isoniazide in the Center Hospital of Changsha from January 1 ,2013 to September 30 ,2014 .Methods A total of 1 031 strains with positive culture result and identified as strains of Mycobacterium tuberculosis were used absolute concentration method to do the conventional drug susceptibility ,and detected rifampicin and isoniazide resistance gene including rpoB ,katG and inhA gene locus mutation by chip technology ,the results of two methods were compared using card square test statistics .Results By gene chip method ,the sensitive strain of rifampicin was 896 ,the drug-resistant strains was 135 ,the sensitive strains of isoniazide was 901 strains ,130 drug-resistant strains .Compared with the absolute concentration method ,resistance chip detection results were consistent with rifampicin resistant strains 1 011 strains(including 894 drug-resistant strains ,and 117 sensitive strains) ,the coincidence rate was 98 .00% ,consistent with isoniazideresistant strains 1 005 strains(including 890 drug-resistant strains ,115 sensitive strains) ,the coincidence rate was 97 .48% .The most common spot of rifampin resistance related mutations of rpoB gene was 531TCG to TTG ,accounted for 51 .11% ,followed by 526CAC→TAC ,accounted for 10 .37% ,11 strains with 526TCG to TTG ,accounted for 8 .15% .Isoniazid re-sistance was caused by mutations in katG315AGC→ACC resistant strains ,accounted for 83 .85% ,inhA-15C→T mutations accoun-ted for only 12 .30% .Conclusion The results of gene chip method is highly consistent with that of absolute concentration method , could be a fast and effective method for screening rifampicin and isoniazide ,the resistant gene of Mycobacterium tuberculosis to rif-ampicin and isoniazide almost mutate in rpoB531 ,526 and katG315 in Changsha .

AIM: To purify and refold the inclusion body of a human anti-HBsAg scFv with a 6?His tag, and to determine the affinity constant of the purified recombinant product.METHODS: Solubilizing in buffers containing urea or guanidine hydrochloride (GuHCl), the inclusion body was purified by IMAC, and then refolded by dialysis against urea or GuHCl, at the same time, Ni 2+ charged chelate column was utilized for in situ refolding. The affinity constant of the refolded scFv, polished by immune-affinity chromatography, was determined by non-competitive ELISA. RESULTS: The refolded scFv with highest specific bioactivity was produced by dialysis against GuHCl. Under this condition, the recovery of target protein reached (61.08?1 45)%. The affinity constant of the polished scFv was confirmed to be(2.30?0.32) ?10 7 L/mol. CONCLUSION: The inclusion body studied in this paper can be refolded efficiently under optimal dialysis condition in vitro . The antigen-binding property of this recombinant scFv is not affected by the purification tag fused to the N terminal of the protein.

Objective: To investigate whether grain-sized moxibustion at Xinshu (BL15) and Shenshu (BL23) can alleviate cognitive decline and other pathologic features in early-stage Alzheimer disease (AD) using transgenic mice with 5 familial AD mutations (5XFAD). Methods: The genotype of transgenic mice was detected by polymerase chain reaction. A total of 40 transgenic mice (1.5 months old) were randomly and equally allocated to an AD model group (5XFAD group) or a grain-sized moxibustion group (5XFAD + GM group), with 20 wild-type (WT) mice (C57BL/6J) serving as the normal control group (WT group). Mice in the 5XFAD + GM group were treated by grain-sized moxibustion at bilateral Xinshu (BL15) and Shenshu (BL23). Mice in the WT group and 5XFAD group received no treatment but were restrained to ensure exposure to a similar experimental condition. Cognitive function and memory were assessed with the Morris water maze and Y-maze tests. The amyloid β 40 (Aβ40) and amyloid β 42 (Aβ42) levels in the brain were evaluated by enzyme-linked immunosorbent assay; amyloid plaque deposition in brain tissue sections was detected by thioflavin-S staining; the expression of glial fibrillary acidic protein (GFAP), cluster of differentiation 11b (CD11b), brain-derived neurotrophic factor (BDNF), and choline acetyltransferase (ChAT) in the hippocampus and prefrontal cortex was analyzed by immunohistochemistry. Results: In the Morris water maze test, compared with the 5XFAD group, mice in the 5XFAD + GM group had a shorter escape latency and more target area crossings and spent more time in the target quadrant (P<0.05). In the Y-maze test, compared with the 5XFAD group, the number of training times of the 5XFAD + GM group was significantly decreased (P<0.05), together with more correct responses (P<0.05). Compared with the 5XFAD group, the levels of Aβ40 and Aβ42 in the brain tissue of the 5XFAD + GM group were significantly lower (P<0.05); in the hippocampus and prefrontal cortex, the total number of amyloid β plaque deposition were significantly lower (P<0.05); the expression levels of GFAP and CD11b were significantly reduced (P<0.05); and the expression levels of ChAT and BDNF were significantly increased (P<0.05).Conclusion: Grain-sized moxibustion at Xinshu (BL15) and Shenshu (BL23) greatly improves learning and memory functions, decreases the levels of Aβ40 and Aβ42, inhibits amyloid β plaque deposition, decreases the expression of GFAP and CD11b, and increases the expression of ChAT and BDNF in AD mice to inhibit the progression of AD.

Objective To study the effect of menopause status and the time of taking tamoxifen (TAM) on endometrial lesions after breast cancer surgery. Methods A total of 330 patients with postoperative vaginal irregular bleeding after breast cancer surgery or endometrial lesions after B ultrasonic from August 2007 to August 2017 in Northern Jiangsu People 's Hospital were retrospectively analyzed, including 180 cases of taking TAM treatment (medicine-taking group), and 150 cases of not taking TAM treatment (non medicine-taking group). The patients were also divided into the menopause group and the premenopausal group. According to the time of taking TAM, the patients were divided into < 2 years group, 2-5 years group and > 5 years group. Chi-square and Fisher test were used to compare the differences. Results The endometrial lesions incidence in the medicine-taking group was higher than that in the non medicine-taking group [84.44 ％ (152/180) vs. 56.00％(84/150);χ2=51.701, P=0.000]. The endometrial lesions rate in the menopause group was higher than that in the premenopause group [medicine-taking group: 69.70 ％ (46/66) vs. 92.98 ％ (106/114), χ2= 17.254, P= 0.000; non medicine-taking group: 46.15 ％ (35/65) vs. 63.53 ％(54/85), χ2 = 4.513, P= 0.034]. For the patients in the menopause group and the premenopause group, the incidence of endometrial lesions for those who took medicine for>5 years [96.00％(48/50), 85.19％(23/27)] was higher than that in the<2 years group and 2-5 years group [78.26％(18/23), 42.86％(6/14);95.12％(39/41), 72.00％(18/25) respectively], and there were statistical differences (χ2=7.619, P=0.022;χ2= 8.070, P= 0.018). The menopause was not correlated with staging, muscular lawyer infiltration and lymph metastasis postoperative (P> 0.05), but with the type of endometrial cancer (P= 0.013); the length of taking medicine was related with the type of endometrial cancer and the lymph metastasis (P=0.027). With the prolonged time of medicine-taking for postmenopause patients, the incidence of type Ⅱendometrial cancer and positive rate of lymph metastasis were also increased. Conclusions Taking TAM after surgery for breast cancer patients increases the risk of endometrial lesions. The longer the patients take the medicine, the greater risk of the lesions take, and the worse the pathological, histological type and prognosis of endometrial carcinoma are, which is more obvious for postmenopausal women who take TAM for more than 5 years.

Objective:To construct and validate a theoretical model of residents′ willingness to participate in tiered medical care based on harmonious management theory, providing reference for promoting the tiered medical care system and aiding governmental decision-making.Methods:Based on the harmonious management theory and literature review, a model capturing residents′ propensity to engage in tiered medical care was formulated. Using convenience sampling method, a questionnaire survey was conducted on 2 067 residents from 24 communities in Zhejiang province from April to May 2022. Descriptive statistical analysis was conducted on the survey results, and multilevel linear regression and structural equation modeling were used to analyze the driving mechanism of residents′ willingness to participate in tiered medical care.Results:The willingness rate of residents to participate in tiered medical care was 69.7%, which was at an average level. Regression insights highlighted the positive influence of policy support perception ( β=0.170, P<0.01), awareness of management mechanisms ( β=0.093, P<0.01), cognitive attitudes ( β=0.102, P<0.01), and trust levels ( β=0.244, P<0.01) on residents′ participation willingness. In contrast, resource allocation perceptions lacked a significant effect ( β=0.065, P>0.05). The structural equation model revealed that cognitive attitudes played a mediating role in the " policy system perception → participation willingness" and " management mechanism perception → participation willingness" pathways, with effect sizes of 0.030 and 0.039, respectively. Trust levels also mediated these paths, with effect sizes of 0.039 and 0.045, and entirely mediated the " resource allocation perception → participation willingness" path, registering an effect size of 0.053. Conclusions:The harmonious management theory can be used to explain the formation mechanism of residents′ willingness to participate in tiered medical care. The government and medical institutions urgently need to further improve residents′ awareness of tiered medical care, focus on enhancing residents′ trust, and further improve policies and management measures such as financial investment, medical insurance reimbursement, and referral systems.

OBJECTIVETo achieve early diagnosis for inheritable hearing loss and determine carrier rate of deafness causing gene mutations in order to provide information for premarital, prenatal and postnatal genetic counseling.

METHODSA total of 17 000 dried heel blood spots of normal newborns in Chengdu were collected with informed consent obtained from their parents. Genomic DNA was extracted from dried blood spots using Qiagen DNA extraction kits. Microarrays with 9 common mutation loci of 4 deafness-associated genes in Chinese population were used. Nine hot mutations including GJB2 (35delG, 176del16, 235delC and 299delAT), GJB3 (538C> T), SLC26A4 (IVS 7-2A> G, 2168A> G), and mitochondrial DNA 12S rRNA (1555A> G, 1494C> T) were detected by PCR amplification and microarray hybridization. Mutations detected by microarray were verified by Sanger DNA sequencing.

RESULTSOf the 17 000 new-borns, 542 neonates had mutations of the 4 genes. Heterozygous mutations of GJB2, at 235delC, 299delAT, and 176del16 were identified in 254, 55, and 15 newborns, respectively. Two newborns had homozygous mutation of GJB2, 235delC. Heterozygous mutations at 538C> T of GJB3, 2168A> G and IVS 7-2A> G of SLC26A4 were found in 23, 17 and 128 newborns, respectively. For mutation analysis of mitochondrial DNA 12S rRNA, 1494C> T and 1555A> G were homogeneous mutations in 4 and 42 neonates, respectively. In addition, 6 complexity mutations were detected, which demonstrated that one newborn had heterozygous mutations at GJB2 235delC and SLC26A4, IVS7-2A> G, one had heterozygous mutation GJB2 235delC and 12S rRNA homogeneous mutation, 1555 A> G, one heterozygous mutations at GJB2, 299delAT, and GJB3, 538C> T, one at GJB2, 299delAT and 12S rRNA, 1555 A> G, two at GJB2, 299delAT, and SLC26A4, IVS7-2A> G. All mutations as above were confirmed by DNA sequencing.

CONCLUSIONThe total mutation carrier rate of the 4 deafness genes is 3.19% in healthy newborns at Chengdu. Mutations of GJB2 and SLAC26A4 are major ones (86.5% of total). The mutation rate of mitochondrial DNA 12S rRNA is 2.71‰, which may have deafness induced by aminoglycoside antibiotics. Newborn screening for mutation of genes related to hereditary deafness plays an important role in the early detection and proper management for neonatal deafness as well as genetic counseling for premarital, prenatal and postnatal diagnosis.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; DNA, Mitochondrial ; chemistry ; genetics ; Deafness ; diagnosis ; ethnology ; genetics ; Dried Blood Spot Testing ; Genetic Predisposition to Disease ; ethnology ; genetics ; Genetic Testing ; methods ; Humans ; Infant, Newborn ; Membrane Transport Proteins ; genetics ; Microarray Analysis ; methods ; Mutation ; Neonatal Screening ; methods ; RNA, Ribosomal ; genetics