ObjectiveTo investigate the neuroprotective effects of Tiaoseng decoction on a perimenopausal depression （PMD） rat model and to examine its regulatory influence on the estrogen receptor β （ERβ）/monoamine oxidase A （MAOA）/c-Jun N-terminal kinase （JNK） signaling pathway， thereby elucidating its potential mechanisms of action. MethodsForty-eight female Sprague-Dawley （SD） rats were divided into a sham group， a model group， a 17β-estradiol （E₂） group （2.5 × 10^-5 g·kg^-1）， and low-， medium-， and high-dose Tiaoseng decoction groups （9.69， 19.37， 38.74 g·kg^-1） by using a random number table method， with eight rats in each group. The PMD model was replicated using ovariectomy （OVX） combined with chronic unpredictable mild stress （CUMS） and was treated continuously with 17β-E₂ and different doses of Tiaoseng decoction for 28 d， once a day. Depressive-like behaviors were assessed using the sucrose preference test， open-field test， and forced swim test. Histopathological changes in the prefrontal cortex were examined using hematoxylin-eosin （HE） and Nissl staining. Enzyme-linked immunosorbent assay （ELISA） was performed to measure serum levels of E₂， luteinizing hormone （LH）， and follicle-stimulating hormone （FSH）， as well as reactive oxygen species （ROS）， malondialdehyde （MDA）， and superoxide dismutase （SOD） levels in the prefrontal cortex. Mitochondrial ultrastructure of prefrontal cortex neurons was observed via transmission electron microscopy. Terminal deoxynucleotidyl transferase dUTP nick-end labeling （TUNEL） was used to detect neuronal apoptosis in the prefrontal cortex. Immunohistochemistry （IHC） was applied to analyze the protein expression of brain-derived neurotrophic factor （BDNF）， postsynaptic density protein95 （PSD95）， and synaptophysin （SYP） in the prefrontal cortex. Immunofluorescence （IF） was conducted to evaluate the average fluorescence intensity of ERβ and MAOA in the prefrontal cortex. Western blot and real-time quantitative polymerase chain reaction （Real-time PCR） were used to determine the protein and mRNA expression levels of key molecules in the ERβ/MAOA/JNK signaling pathway in the prefrontal cortex. ResultsCompared with the sham group， the model group had significantly reduced sugar-water preference index， total distance traveled， average speed， and activity time in the central region in the open field experiment， E₂ content， and SOD content （P<0.01） and significantly reduced BDNF， PSD95， SYP， and ERβ expression and B-cell lymphoma-2 （Bcl-2）/Bcl-2-associated X protein （Bax） ratio （P<0.01）. Additionally， the model group exhibited severe histopathological damage， disrupted mitochondrial structure， cristae disappearance or fracture， swelling， and deformation in the prefrontal lobe. The immobilization time of forced swimming， TUNEL positivity， LH， FSH， MDA， ROS， MAOA， Caspase-3， p-JNK/JNK， and p-c-Jun/c-Jun were significantly increased in the model group compared with the sham group （P<0.01）. Compared with the model group， the low-， medium-， and high-dose Tiaoseng decoction groups and the 17β-E₂ group had increased sugar-water preference index， total distance traveled， average speed， and activity time in the central region in the open-field test， E₂ content， and SOD content （P<0.05， P<0.01） and elevated BDNF， PSD95， SYP， and ERβ expression and Bcl-2/Bax ratio （P<0.05， P<0.01）. In addition， histopathological damage to the prefrontal lobe was improved to different degrees， and mitochondrial structure was gradually repaired. The immobilization time of forced swimming， TUNEL positivity， LH， FSH， MDA， ROS， MAOA， Caspase-3， p-JNK/JNK and p-c-Jun/c-Jun were significantly reduced in the low-， medium-， and high-dose Tiaoseng decoction groups and the 17β-E₂ group compared with the model group （P<0.05， P<0.01）. ConclusionTiaoseng decoction has significant neuroprotective effects on PMD model rats， which can alleviate perimenopausal depressive disorder by inhibiting oxidative stress， attenuating neuronal apoptosis， and restore synaptic plasticity via ERβ/MAOA/JNK signaling pathway regulation.

Objective To investigate the regional distribution of positive results of early screening for women of child-bearing agein the six provinces of southern China(Guizhou,Guangxi,Yunnan,Sichuan, Chongqing and Hunan).Methods The screening of all were performed by red blood cell related parameters、quantitative analysis of hemoglobin、Osmotic fragility test of erythrocyte incubation(one tube method)、dena-tured hemoglobin inclusion body test among 222 645 peoples reproductive age in the six provinces of southern China,and statistical analysis of abnormal screening in different provinces(cities)was carried out.Results A-mong 222 645 cases in the six provinces(cities),abnormal samples were detected in 32074 cases,and the abnor-mal detection rate was 14.39%.The total positive detection rate from high to low according to the order of Guangxi(24.87%),Sichuan(17.53%),Yunnan(14.76%),Hunan(11.53%),Chongqing(10.69%)and Guizhou(8.61%).Among the six provinces(city),the total α and β thalassemia screening positive rate were 16.37%,17.81%,the specific distribution(α/β):Guangxi(7.06% /5.89%),Sichuan(2.74% /3.02%),Hunan (1.87% /2.32%),Yunnan(1.56% /1.57%),Guizhou(1.85% /3.04%)and Chongqing(1.29% /1.97%).The total α and β thalassemia screening suspected positive rate were 50.56%,3.25%,the specific distribution(α/β):Guangxi(11.59% /0.33%),Sichuan(11.27% /0.50%),(9.58% /2.05%)in Yunnan,Chongqing(7.35% /0.08%),(7.12% /0.22%)of Hunan and Guizhou(3.65% /0.07%).Conclusion The six southern provinces (city)positive screening rate distribution is different,the highest in Guangxi,followed by Sichuan,Guizhou is the lowest;suggestions will flow into the southern city and high malaria area included in in screening of thalas-semia.The four-way experiment has a higher positive rate of screening for thalassemia,and screening out the number of abnormal hemoglobin disease,and has great value of reduce the misdiagnosis.In order to further re-duce the misdiagnosis,suggested that serum iron detection into thalassemia screening.Strengthen the screen-ing efforts for thalassemia in high risk families,reducing the birth rate of thalassemia major,improve popula-tion quality,reduce the economic burden of the family and the country.

Objective To investigate the DNA staining efficiencies of 4 kinds of nucleic acid fluorescent dyes,including EB,SYBR Green Ⅰ,Gold View and AO,in single-cell gel electrophoresis (SCGE) assay,and the possibility to use a new nucleic acid fluorescent dye instead of EB.Methods The peripheral blood lymphocytes from healthy individuals were isolated and treated with 0,20,40,60 and 80 μg/mL of H2O2,respectively.Then,the DNA damages of lymphocytes were detected by the neutral SCGE assay.The DNA was stained with EB,SYBR Green Ⅰ,Gold View and AO dyes,respectively,and the staining results were observed and compared under a fluorescence microscope.In addition,the percentages of tail DNA (％ Tail DNA) from different staining methods were analyzed and compared.Results The results of SCGE showed that SYBR Green Ⅰ,Gold View and AO staining could well reflect the DNA damages of lymphocytes,and that the optimal concentrations for SYBR Green Ⅰ,Gold View and AO were 1 ×-5 ×,2 ×-5 × and 2-5 μg/mL,respectively.The regression coefficients for EB,SYBR Green Ⅰ,Gold View and AO were 2.71,2.81,2.73 and 2.75,respectively,which indicated that there was consistent dyeing effect between them.The results of ％ Tail DNA were stable with in 48 hours,and there was no significant difference between the 4 fluoresent dyes (P ＞ O.05).The inter-assay coefficients of variation (CVs) of SYBR Green Ⅰ,Gold View and AO were 6.92％,7.10％ and 8.25％,respectively,which were superior to that of EB (8.35％).The intra-assay CVs of SYBR Green Ⅰ and Gold View were 3.07％ and 2.74％,respectively,which were superior to that of EB (3.59％).Conclusion SYBR Green Ⅰ,Gold View and AO may be used for the nucleic acid fluorescent staining,and especially Gold View is more suitable for instead of EB in SCGE.

Objective To investigate the expressions of phosphorylated H2AX (γH2AX) and p53-binding protein 1 (53BP1) in DNA oxidative damage of human bronchial epithelial (HBE) cells.Methods The HBE cells were treated with 0,25,50,100,200,400 μmol/L of hydrogen peroxide (H2O2) for 1 hour,respectively,and their DNA oxidative damages displaying as double-strand breaks (DSBs) were induced.The viability and apoptosis of HBE cells were measured by the CCK-8 method and flow cytometry,respectively.The expression status of γH2AX and 53BP1 in nucleus of HBE cells was observed by a fluorescence microscope.The expression levels of γH2AX,53BP1 and BRCA1 were determined by western blot.Results Compared with the control (0 μmol/L of H2O2),the via bility of HBE cells treated with 25 μmol/L of H2O2 (1.07 ±0.01) increased,while those with 50,100,200,400 μmol/L of H2O2 (0.97 ± 0.01,0.96 ± 0.01,0.95 ± 0.01,0.94 ± 0.01) decreased significantly (F =50.35,P ＜ 0.01).The apoptosis rates of HBE cells treated with 50,100,200,400 μ mol/L of H2O2 ([7.54 ± 0.57] ％,[7.84 ± 0.68] ％,[8.40 ± 0.50] ％ and [14.03 ± 1.03] ％) were significantly higher than that with 0 μmol/L of H2O2 ([4.65 ± 0.32] ％,F =35.879,P ＜ 0.01).Compared with the control (0 μmol/L of H2O2),the average fluorescence intensity of γH2AX in nucleus of HBE cells treated with 25,50,100,200,400 μmol/L of H2O2 increased significantly (F =223.97,P ＜ 0.01),while those of 53BP1 in nucleus of HBE cells treated with 50,100,200,400 μmol/L of H2O2 decreased significantly (F =117.78,P ＜ 0.01).The results of western blot showed that the expres sion levels of γH2AX increased with the increase of H2O2 concentration,while that of 53BP1 and BRCA1 was on the contrary (F =96.20,21.92 and 11.55,respectively,P ＜0.01).Conclusion In the oxidative damage of HBE cells induced by H2O2,γH2AX may be used as a marker of DNA oxidative damage,while the decreased expression of 53BP1 suggests that other mechanisms to repair the DNA damage sites may exist.

Objective: To investigate the efficacy and safety of IA regimen which contains idarubicin (IDA) 8 mg/m², 10 mg/m² or 12 mg/m² as induction chemotherapy for adult patients with de-novo acute myeloid leukemia (AML) . Methods: A total of 1 215 newly diagnosed adult AML patients, ranging from May 2011 to March 2015 in the First Affiliated Hospital of Soochow University and other 36 clinical blood centers in China were enrolled in the multicenter, single-blind, non-randomized, clinical controlled study. To compare the response rate of complete remission (CR) , adverse events between different dose idarubicin combined with cytarabine (100 mg/m²) as induction chemotherapy in newly diagnosed patients of adult AML. Results: Of 1 207 evaluable AML patients were assigned to this analysis of CR rate. The CR rates of IDA 8 mg/m² group, IDA 10 mg/m² group and IDA 12 mg/m² group were 73.6% (215/292) , 84.1% (662/787) and 86.7% (111/128) , respectively (P<0.001) . After adjusted for age, blast ratio of bone marrow, FAB classification and risk stratification, the odds ratios (95% CI) of IDA 10 mg/m² group and IDA 12 mg/m² group were 0.49 (0.34-0.70) and 0.36 (0.18-0.71) , as compared with the IDA 8 mg/m² group (P<0.001, P=0.003) . In the intermediate and favorable groups, CR rates was 76.5% (163/213) , 86.9% (506/582) and 86.1% (68/79) in different doses of IDA (P=0.007) . Interestingly, IA regimen with IDA 10 mg/m² was the only beneficial factor affecting CR in this group after adjusted for age, blast ratio of bone marrow and FAB classification[OR=0.47 (95% CI 0.31-0.71) , P<0.001]. CR rates in adverse group was 50.0% (18/36) , 60.6% (43/71) and 81.8% (18/22) respectively (P=0.089) . However, the odds ratios (95% CI) of IDA 12 mg/m² when compared with the IDA 8 mg/m² was 0.22 (0.06-0.80) , after adjusted for age, blast ratio of bone marrow and FAB classification. The median time (days) of neutrophil count less than 0.5×10⁹/L in IDA 8 mg/m² group, IDA 10 mg/m² group and IDA 12 mg/m² group were 14 (11-18) , 15 (11-20) and 18 (14-22) , respectively (P=0.012) and of platelet count lower than 20×10⁹/L were 14 (7-17) , 15 (11-20) and 17 (15-21) , respectively (P=0.001) . The incidences of lung infection in the three groups were 9.8%, 13.5% and 25.2%, respectively (P<0.001) . Conclusions For young adult patients (aged 18-60 years) with AML in China, intensifying induction therapy with idarubicin 10 mg/m² is clinically superior to IDA 8 mg/m² and IDA 12 mg/m² in favorable intermediate AML subgroup. However, idarubicin 12 mg/m² is more suitable to adverse AML subgroup.

Laboratory medicine is experiencing rapid development and moving from the edge of medicine stage to the center of medicine.Laboratory medicine has become the component of evidence-based medicine,the pathway to translational medicine and the core of precision medicine.These recent advances have increased the challenges and opportunities for laboratory medicine education.It will be the most urgent task to us how to train innovative personnel, medical laboratory physician, genetic counselor, medical laboratory scientist and personnel needed for personalized medicine.

Objective To establish an inductively coupled plasma‐mass spectrometry (ICP‐MS) method for analysis of elements in peripheral blood of children .Methods A total of 474 healthy children in Hunan area were enrolled in this study ,and six ele‐ments ,including Ca ,Mg ,Fe ,Cu ,Zn and Pb ,in peripheral blood specimens were detected by using ICP‐MS method .Results The levels of elements ,including Ca ,Mg ,Fe ,Cu ,Zn and Pb ,in peripheral blood of healthy children showed skewed distributions ,and no significant differences were found in levels of these elements between male and female children(P>0 .05) .The reference intervals of Ca ,Mg ,Fe ,Cu ,Zn and Pb in peripheral blood of healthy children in this area were 57 .30 -81 .40 mg/L ,30 .40 -44 .80 mg/L , 361 .20-531 .40 mg/L ,848 .10-1 469 .20 μg/L ,2 .68 -6 .54 mg/L and 0 .00 -100 .00 μg/L respectively .Conclusion The ICP‐MS method for simultaneously detecting Ca ,Mg ,Fe ,Cu ,Zn and Pb in peripheral blood of children and reference interval of each ele‐ment are successfully established .

Objective:To improve the immunoblotting of immunoprecipitated proteins and decrease the interference of immunoprecipitation antibody in the interaction of endogenous proteins. Methods:Transient transfect cells with fusion protein expression vector containing the targeted S5b gene and the FLAG tag, the transfected cells or untransfected cells were harvested to study the exogenous or endogenous protein interaction. The total cell lysate was immunoprecipitated by specific antibody. Then the eluted immunocomplex was separated by SDS-PAGE, and the TrueBlot antibody or conventional antibody was used as the secondary antibody for immunoblotting detection of S5b and its partner (Rpt1 and Rpt2). Results:Clear immunoblotting bands for S5b, Rpt1 and Rpt2 were obtained. Conclusion:TrueBlot antibody prefers the immunoblot antibody to immunoprecipitation antibody, and decreases the interruption of immunoprecipitation antibody to display clear protein band.

OBJECTIVE: To develop a rapid detection method for microalbuminuria with size-exclusion high performance liquid chromatography. METHODS: With a new mobile phase, samples were injected onto an Agilent Zorbax GF-250 size exclusion chromatographic column. The method was evaluated and urine albumin of 56 diabetic patients was analyzed. RESULTS: The mobile phase containing 0.1% formic acid and acetonitrile, with 20 μL sample size, 1 mL/min flow rate, 205 nm detection wavelength. The retention time of albumin both in human serum and urine was 1.7 min. The linear range was 5-2000 mg/L. The lower limit of measurement was 2 mg/L. The intra-assay coefficient of variation and the inter-assay coefficient of variation were 3.98% and 4.05% (20 mg/L), 3.55% and 3.60% (200 mg/L), 4.65% and 4.74% (2000 mg/L), respectively. Recovery rates were 95.3%, 98.1%, and 97.2%. Microalbuminuria was detected in 30 samples by high performance liquid chromatography and 15 samples by immunoturbidimetry from 56 patients with diabetic mellitus. CONCLUSION A fast and high sensitivity method, namely size-exclusion high performance liquid chromatography, with mobile phase containing 0.1% formic acid and acetonitrile has been established to analyze microalbuminuria, which can detect more microalbuminuria than other methods and is suitable for clinical routine measurement.
Albuminuria ; diagnosis ; Chromatography, High Pressure Liquid ; Diabetes Mellitus ; urine ; Humans

Objective Analysing the potential relationship of mismatch repair protein MLH3 gene C2531T mutation with oligospermia and azoospermia cases. Methods Using MLH3 C2531T as an example to establish a method of tetra-primer amplification refractory mutation system-PCR (4P-ARMS-PCR); We chose 81 cases of oligospermia, azoospermia patients and 80 normal controls, detecting MLH3 C2531T both using 4P-ARMS-PCR and PCR-RFLP; randomly to select 20% samples to sequence. Results The results suggested higher prevalence of CT + TT genotypes in patients as compared to the controls and association of T allele with disease (P <0. 01); PCR-RFLP results , 4P-ARMS-PCR results and sequence results are fully consistent. Conclusion 4P-ARMS-PCR can be a fast, easy-operating and accurate technology for detection of gene mutation of single nucleotide polymorphism (SNP) ; MLH3 C2531T polymorphism is related to oligospermia and azoospermia.