β-thalassemia is one of the most common monogenic genetic disorders in the world,and represents a major public health problem among humans.This review descrihes the latest technical development of molecular diagnosis of β-thalassemia from the perspective of clinical settings.Related techniques including reverse dot blot,minisequencing,real-time PCR,and non-invasive prenatal diagnosis,etc.are discussed regarding their principles,features and trends.

【Objective】To study the correlation of human leucocyte antigen(HLA) class Ⅱ allele with spleen-stomach damp-heat(SSDH) syndrome.【Methods】From those people with native place of Guangdong,16 healthy volunteers served as the normal control and 46 patients with chronic superficial gastritis or digestive ulceration were enrolled into the observation.Of 46 patients,26 were classified into SSDH and 20 into spleen deficiency(SD) syndrome.HLA-DRB1,HLA-DQA1,HLA-DQB1,and HLADPB1 alleles of HLA class Ⅱ from whole blood sample were analyzed by the method of polymerase chain reaction-sequence specific primers(PCR-SSP).【Results】The gene frequency of DQA1*0103 in SSDH group was remarkably higher than that in the normal control group(P

AIM: To construct E. coli expression plasmid of recombinant human NDPK-A with a 6?His tag, optimize the expression condition and identify the activity of the product. METHODS: nm23-H1 was subcloned from plasmid pBVNMH1 to pQE40 which contain 6?His purification tag. The expression condition was modulated in grades to get the optimal expression. We purified protein with the Ni+-NTA affinity chromatography column, identified the immunogenicity of the product with Western blot, and measured the kinases activity with HPLC. In addition, angiogenesis inhibition activity of rhNDPK was identified by CAM. RESULTS: The sequence of nm23-H1 subclone in pQE40 was exactly correct. The expression rate of rhNDPK-A was 49 6%. Purified rhNDPK-A specially recognized the antiserum of NDPK-A. It also inhibited angiogenesis. CONCLUSION: PQE-nm23H1 containing 6?His can express target protein at high level. This purification method is simple than other methods, and the product has the same activity as natural human NDPK-A.

Objective To investigate the expression of TLR4 mRNA and NF-κB p65 in colorectal carcinomas and adjacent normal colon tissue, and evaluate their roles in the pathogenesis and development in colorectal carcinoma. Methods Sixty-three colorectal carcinoma samples and respective adjacent normal colon tissue samples ( well differentiated : 23 cases; moderately differentiated: 17 cases; poorly differentiated:20 cases; other differentiated type: 3 cases; lymph node metastasis: 27 cases; no lymph node metastasis:36 cases; Dukes A: 18cases;Dukes B: 14 cases Dukes C: 22 cases; Dukes D: 9 cases) were collected. The expression of TLR4 mRNA in colorectal carcinomas and adjacent tissue were detected by RT-PCR. The expression of NF-κB p65 was detected by WB. Results The expression of TLR4 mRNA in colorectal carcinomas and adjacent tissue were 86.42 ± 15.16 and 32.74 ± 9.44. It was significantly higher in carcinoma tissue than that in adjacent tissue ( t = 22.354, P ＜ 0.01 ). The expression of TLR4 mRNA in well, moderately and poorly differentiated coiorectal carcinomas were 69.58 ± 11.27, 64.57 ± 13.91 and 97.12 ± 15.44 respectively. TLR4 mRNA in poorly differentiated colorectal carcinomas was significantly higher than that in well, moderately differentiated ones ( t = 11.304 and 12.223, P ＜ 0.01 ). There was no difference between lymph node metastatic carcinomas ( 89.91 ± 13.33 ) and carcinomas without metastasis (81.16±13.59,t =0.959,P＞0.05). The expression of TLR4 mRNA in the Dukes A stage tumors (59.05±11.66) was lower than that in Dukes B(90.34 ±0.08),C(91.41 ± 15.21), D(101.46 ±17.43), respectively ( t = 8.708,9.664,9.525, P ＜ 0.05 ). The expression of NF-κB p65 in colorectal carcinoma(0.63 ±0.11) was significant higher than that in adjacent tissue(0.34 ±0.08,t = 18.266,P ＜0.01 ). The expression of NF-κB p65 in well, moderately and poorly differentiated colorectal carcinomas were 0.46 ± 0.09, 0.72 ± 0.11 and 0.77 ± 0.14, respectively. The experssion of NF-κB p65 in well differentiated colorectal carcinomas was obviously lower than the woderately and poorly differentiated carcinomas (t = 11.223 and 10.875, P ＜0.01 ). There was significant difference between the expression of p65 in lymph node metastatic carcinomas(0.82 ± 0.17) and non-metastatic carcinomas(0.57 ± 0.12, t =18.269,P＜0.05). The expression of NF-κB p65 in Dukes A colorectal carcinomas (0.39 ± 0.06) was lower compared with the Dukes B(0.72 ±0.12), C(0.69 ±0.14) and D carcinomas(0.76 ±0.13,t =10.442, 9.889 and 9.721, P ＜ 0.01 ). Conclusions The enhanced expression of TLR4/NF-κB p65 are closely associate with clinical stage and pathologic grade. NF-κB p65 may be an molecular marker of lymph node metastatic. The increased expression of TLR4/NF-κB p65 promote the pathogenesis and development of colorectal carcinoma.

As injury to distal tibiofibular syndesmosis is commonly associated with ankle fractures,it calls for timely and correct diagnosis and treatment because it might result in chronic pain and ankle instability.Its severe cases can lead to traumatic arthritis.Stable injury to distal tibiofibular syndesmosis can be treated conservatively,but internal fixation is mostly recommended for severe cases of the injury.Internal fixation with metal screws usually requires a secondary surgery for removal of the implants but application of bioabsorbable screws overcomes this trouble and avoids a secondary surgery and its related complications.This article reviews the advantages of bio-absorbable screws,choice of bioabsorbable materials and screws,therapeutic outcomes and adverse reactions in the treatment of distal tibiofibular syndesmotic injury.

Objective To establish a mouse model of systemic C. albicans infection by oral inoculation of the pathogen and observe the proliferation and distribution of C. albicans in vivo tissues. Methods Male ICR mice(n=46) were used as the experiment group(n=40) and blank group (n=6). Cotton swabs with C. albicans were used to infect the mice (7 × 106 CFU/mL), and the blank group with saline. The mice of the experiment group were randomly divided into two groups:model group A for clinical assessment (n=20) and model group B for tissue fungal burden detection (n=20). Clinical score, survival and autopsy were carried out among the model group A. Five mice were randomly killed from the model group B at 3 d, 5 d and7 d after infection, respectively ( blank group killed 2 mice each time) . Microbial load tablet method was used to detect the tissue fungal burdens in different tissues, meanwhile samples of tongue, esophagus, stomach, liver, kidney, lung of infected mice were taken for pathological examination. Results White spot appeared on the surface of tongue since 3 d postinfection and increased with time and finally caused death. The mortality reached over 50% at 5 d. C. albicans was not only detected from the tongue (87?5%), stomach (87?5%), liver (54?5%), kidney (50?5%), lung (20%) and heart (4%), but also was microscopically seen mycelia proliferation in the tongue, stomach, liver, and kidney , yet not seen in the control group, showing that C. albicans caused disseminated systemic infection through mucosal infection in mice. Conclusions C. albicans can induce opportunistic systemic infection by breakthrough the mucosal immune barrier, so as to increase the infection to death.

Objective To explore the influencing factors of late diagnosis for newly diagnosed HIV/AIDS positive patients in Guangxi in 2015.Methods The CD4 + T lymphocytes count which was first detection for newly diagnosed HIV/AIDS positive patients in Guangxi during 2015 was collected.Data were statistically analyzed.Results We collected 8 586 newly diagnosed HIV/AIDS whose median CD4+ T lymphocytes counts was 237.5 cells/μl,and 43.12％ of them had less than 200 cells/μl.Gender,age,occupation,marriage,nation,education,route of transmission,types of testing and region had effects on late HIV diagnosis(all P ＜ 0.05).Logistic analysis found that risk factors associated with the late diagnosis of HIV were male(OR =1.851,95％ CI:1.673-2.048),migrant worker (OR =1.387,95％ CI:1.242-1.549),education below middle and secondary school(OR =1.619,95％ CI:1.400-1.873),currently married(OR =1.207,95％ CI:1.075-1.354),divorced or widowed(OR =1.508,95％ CI:1.309-1.738).Voluntary testing was a protective factor.Conclusions The prevalence the late diagnosis of HIV was high in Guangxi in 2015,it is crucial for related departments to enhance the testing and screening effort for HIV/AIDS.

Objective To investigate the relationship between carotid atherosclerotic plaque and multiple risk factors of angiocardiopathy,and to evaluate the injuries caused by different risk factors to subclinical target organ to control the general risk factors of angiocardiopathy.Methods Four hundred and twenty six outpatients and impatients,treated in our hospital from May 2007 to May 2009 with the results of color ultrasonic examination,were divided into carotid atherosclerotic plaque group(284 cases) and no carotid atherosclerotic plaque group( 142 cases).The clinical information including their age,body mass index,smoking condition,past medical history such as hypertension,diabetes mellitus and hyperlipoidemia were recorded,and the levels of total cholesterol(T C),high density lipoprotein cholesterol( HDL-C),low density lipoprotein cholesterol(LDL-C),triglyceride (TG),lipoprotein ( a ) ( LP (a) ),apolipoprotein A - 1 ( Apo A 1 ),apolipoprotein B ( Apo B ),highsensitivity C-reactive protein( hs-CRP),homocysteine ( HCY),microalbuminuria( MAU ) and uricacid(UA) were determined by lab tests.The independent variable and univariable data were processed and analyzed statistically to find out the risk factors of carotid atherosclerotic plaque.Results Age and drinking were significantly correlated with the carotid intima-media wall thickening(IMT) (P ＜ 0.001 ).Overweight,diabetes mellitus,increased LP (a),hyperlipoidemia,age,increased MAU and HCY could independently predict carotid atherosclerosis and plaque formation ( x2 =71.35,38.45,t =3.26,x2 =37.23,t =118.51,6.723 and 3.17respectively,Ps ＜ 0.05 ).The aggregated number of the risk factors was correlated to IMT and carotid atherosclerotic plaque ( P =0.0001 ).Conclusion Age,drinking,overweight,diabetes mellitus,increased LP (a),hyperlipoidemia,MAU and HCY are risk factors of carotid atherosclerosis and plaque formation,and the contribution of each factor can multiply and overlap,more risk factors means greater risk.

ObjectiveTo investigate the application value of negative pressure sucking technique in the elderly after nasal septal reconstruction. MethodsForty elderly patients after nasal septal reconstruction were divided into two groups randomly, with twenty patients in each. The cases in observing group received negative pressure sucking technique while those in controlling group received nasal packing with vaseline gauze following the surgery. The amount of bleeding of the first 24 hours after operation, the comfort degrees evaluated by using visual analogue scale (VAS),hematoma of nasal septum, edema in nasal cavity and duration of hospitalization after operation were compared between the two groups. ResultsThe amount of bleeding of the first 24 hours afteroperation was more in observing group than in controlling group[(14.2±8.1)ml vs. (7.2±4.1)ml, P<0.05]. The VAS score and the number of cases who felt discomfortable moderately or severely were significantly less in observing group than in controlling group at 6 h, 12 h and 24 h after operation(all P<0.05). The nasal cavity gap were significantly wider in observing group at the 3rd day after operation(P=0.001), and the duration of hospitalization after operation was significantly shorter in observing group [(4.5±0.9) d] than in controlling group [(5.42±0.9) d](P=0.042)Conclusions Negative pressure sucking technique applied in the elderly after nasal septal reconstruction can significantly relieve the distress of patients, reduce the edema in nasal cavity and shorten the duration of hospitalization, but does not increase the risk of hemorrhage.

OBJECTIVETo evaluate the clinical value of multicolor melting curve analysis(MMCA) for detecting genetic mutations in G6PD deficiency.

METHODSA total of 402 peripheral blood samples(256 males and 146 females) were collected from suspected patients or their relatives at the Prenatal Diagnosis Center of Liuzhou Maternal and Child Health Hospital between March 2012 and May 2012. The samples were screened by G6PD/6PGD quantitative ratio testing. The reliability of the assay was evaluated by multiplex probe melting curve assay(which can detect 16 G6PD mutations) and DNA sequencing through a double blind study.

RESULTSOne hundred seventy cases with G6PD/6PGD ratio < 1.0 and 232 cases with G6PD/6PGD ratio ≥ 1.0 were detected by the enzymological method. DNA sequencing has identified 182 wild type samples, 151 hemizygous mutation samples, 5 female homozygous mutation samples, 54 female heterozygous mutation samples and 10 female double heterozygous mutation samples. Multicolor melting curve analysis has detected 185 wild type samples, 148 hemizygous mutation samples, 5 female homozygous mutation samples, 55 female heterozygous mutation samples and 9 female double heterozygous mutation samples. The specificity and sensitivity of G6PD gene mutation detection by multicolor melting curve analysis were 100%(182/182) and 98.6%(217/220), respectively. The positive predictive value and negative predictive value were 99.5%(216/217) and 98.4%(182/185), respectively, and the Youden's index was 0.986. The concordance rate of the sample detection between the melting curve assay and DNA sequencing was 99.0%(398/402). Twenty-one different genotypes were detected by the multicolor melting curve analysis and 24 different genotypes were detected by DNA sequencing. Four samples containing mutations(c.196T>A or c.406C>T) were not detected by multicolor melting curve analysis, which can be attributed to different technical settings of the two methods.

CONCLUSIONMulticolor melting curve analysis for G6PD gene mutation detection is a simple, rapid, sensitive and specific method, which can be used for clinical diagnosis of G6PD deficiency.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Humans ; Infant ; Infant, Newborn ; Male ; Mutation ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA