The severe infectious diseases caused by virus are occurring with increasing frequency, which poses a rising global threat to human health and life.There are many kinds of viruses that cause a wide variety of viral diseases.The same kind of virus is able to cause different diseases, meanwhile a disease can be caused by different viruses.All these conditions make the relationship between viral pathogens and infection diseases complicated.At present, no effective laboratory detection methods for most of virus infectious diseases are developed.But the rapid development of molecular diagnostic technologies makes it possible for clinical laboratory to detect viral infection diseases rapidly and simultaneously.This review summarized the present and development perspectives of laboratory tests for the diagnosis of clinical virus infections, attempting to give some advices for laboratory diagnosis procedure of viral infections.

TOF-MS has been more and more widely used in clinical and scientific research due to its advantages such as fast and easily operated.Diabetic nephropathy is the most common complications of diabetes.But clinical existing detection meth-ods to some extent have some hysteresis.The applications and developments of TOF-MS coupled with other proteomics technologies provided new ideas and methods for early diagnosis and prognosis of diabetic nephropathy.In this paper,the lat-est research results in nearest years about the application of TOF-MS to diabetic nephropathy will be reviewed.

Objective:To study the effect of endogenous IL-1? on the expression of matrix metalloproteinases (MMPs) and COX2 in dental pulp cells. Methods:Human dental pulp cells were treated with human recombinant IL-1? at 1 nmol/L in serum-free medium for 18 h. Then the cells were collected and total RNA was isolated, MMPs and COX2 mRNA expression was assessed by semiquantitative RT-PCR. Results:IL-1? at 1 nmol/L induced the expression of COX2 and MMP-1 mRNA in human dental pulp cells. Conclusion:IL-1? may contribute to stimulating expression of MMPs and COX2 in the dental pulp during pulpitis.

Objective To evaluate five detection methods for the laboratory diagnosis of Clostridium difficile infection in the hospitals of USA, and explore a sensitive, specific, accuracy and rapid regimen for the early diagnosis of Clostridium difficile infection. Methods A total of 174 stool specimens submitted to the clinical microbiology laboratory for Clostridium difficile testing were separately tested by five methods including toxigenic culture (TGC), Premier Toxin A&B EIA( A/B-EIA), C. Diff Quick Chek Complete( DEIA), BD G eneOhm Cdiff assay(BD-PCR) and Laboratory-developed PCR(LD-PCR). The gold standard of TGC was used as a reference criterion, and the sensitivity, specificity, positive predictive value ( PPV )and negative predictive value (NPV) of A/B-EIA, D-EIA, BD-PCR and LD-PCR assays were determined. Results Among the 174 specimens studied, 24 were defined as true positives for Clostridium difficile infection by TGC assay, giving a positive rate of 13.8% (24/174). In comparison to the standard,the sensitivity, specificity, PPV and NPV were 62.5%, 99.3%, 93.8% and 94.3% for A/B-EIA;66.7%, 98.7%, 88.9% and 94.9% for D-EIA; 83.3%, 98.7%, 90.9% and 97.4% for BD-PCR;79.2%, 93.3%, 65.5% and 96.6% for LD-PCR. Among all tested specimens, 34 were positive by atleast one of five methods, and of which 15 were concordant by all five methods. The D-EIA results were divided into three groups depending on results of GDH and (or) toxins A/B: 18 were positive for both GDH and toxins A/B, 23 were positive for only GDH, and 133 were negative for both GDH and toxins A/B. Of 18 positive specimens by D-EIA assay, all were concordant with results of BD-PCR assay and 16 were agreement with results of TGC assay. Twenty-two of 24 positive specimens by TGC assay were included in 41 specimens that were positive for GDH. Among eight false negative specimens by D-EIA assay, four were differentiated as positive results by BD-PCR. According to the present study, the sensitivity, specificity,PPV and NPV of a two-step detection algorithm in combination with D-EIA and BD-PCR assays were 83.3%, 98.7%, 90.9% and 97.4%, respectively. Conclusions From the point of technological evaluation, BD-PCR is preferable. A two-step detection algorithm combining D-EIA with BD-PCR is proposed for the laboratory diagnosis of Clostridium difficile infection. This algorithm has demonstrated an excellent sensitivity and specificity, as well as decreased test turnaround time and test cost.

Objective By prokaryotic expression and purifying the human bocavirus recombinant protein VP2, to establish the indirect enzyme-linked immunoassay for detection of virus. Methods We amplified the human bocavirus recombinant protein VP2 gene fragments from WHL-1 template by PCR , and cloned into the expression vector pET28a, then conversed into the BL21 (DE3) and expressed the fusion protein detected by Western Blot detection , the obtained the antibody and detected the human bocavirus in serum in Guanghzhou area in healthy people. Results The Recombinant prokaryotic expression identified correct by double enzyme, and it could occur specific reaction with the virus positive serum. The best optimal antigen coating concentration were serum multiples and blocking BSA was 2 mg/mL , 1 ∶ 200 and 1%. The best working dilution of enzyme-labeled secondary antibody was 1 ∶ 4 000. The best working hours was 1h. This detection method had good specificity and reproducibility. The cut-off of the indirect ELISA method was 0.1 and the sensitivity and specificity of the developed ELISA method were 92% and 98% respectively. The coincidence rate of determination results by the developed kit and control kit was 97%. Conclusion The competitive ELISA established by prokaryotic expressing and purifying the human bocavirus protein VP2 protein , provides a basis in detecting the human bocavirus serum antibody.

Objective To evaluate the value of HbA2 level determined by capillary electrophoresis (Hb-CE)in screening and diagnosis of thalassemia.Methods HbA2 level of 249 thalassaemia carriers and 142 healthy controls confirmed by molecular biological detection were determined by Hb-CE method.The thalassaemia carrier subjects were divided into different groups and subgroups according to their results of gene detection.The sensitivity,specificity,accuracy,positive predictive value and negative predictive value for the diagnosis ofα-thalassemia,β-thalassaemia,α,β-thalassaemia were calculated under different HbA2 cut-off value.Results Mean value of HbA2 in healthy controls was (3.03±0.27)%.Mean values of HbA2 inα-thalassemia group and its subgroups of silentα-thalassemia,standardα-thalassemia and hemoglobin H disease were (2.38± 0.55)%,(2.61±0.46)%,(2.47 ± 0.32)% and (1.07 ± 0.17)%,respectively.Mean values of HbA2 inβ-thalassaemia group and itsβ0 subgroup,β+ subgroup were (5.65±0.47)%,(5.71±0.48)% and (5.56±0.43)%.Mean value of HbA2 in compoundαandβ-thalassaemia group was (5.7±0.82)%.Compared with healthy controls,HbA2 level inα-thalassemia group,silentα-thalassemia subgroup,standardα-thalassemia subgroup and hemoglobin H disease group decreased signifi-cantly (t values of 11.73,5.02,12.91 and 33.46,respectively,P<0.01).HbA2 level in hemaglobin H disease was signifi-cantly lower than silent and standardα-thalassemia subgroups (t values of 15.62 and 21.31,respectively,P<0.01),but there were no differences in HbA2 level between silent and standardα-thalassemia subgroups (t=1.50,P>0.05).HbA2 level inβ-thalassaemia group,β0 subgroup,β+ subgroup and compoundαandβ-thalassaemia group increased significantly (t values of 55.12,44.33,38.94 and 9.10,respectively,P<0.01),but there were no differences in HbA2 level betweenβ0 andβ+ subgroups (t=1.79,P>0.05).Of 249 thalassemia carriers,all 124β-thalassaemia carriers were distinguished with ele-vated HbA2 level (>3.5%)determined by Hb-CE and only 57 were distinguished from 117α-thalassemia carriers by Hb-CE.Under the cut-off value of 2.5%,the sensitivity,specificity,positive predictive value,negative predictive value and accu-racy for the diagnosis ofα-thalassemia were 48.72%,97.18%,93.44%,69.70%,75.29%,respectively.Under the cut-off value of 3.5%,they were 100.00%,98.59%,98.41%,100%,and 99.25% for the diagnosis ofβ-thalassaemia,respectively. The analysis of ROC curve showed that the optimal HbA2 cut-off values for diagnosis ofα,β-thalassaemia by capillary elec-trophoresis were 2.8% and 3.7%,respectively.Conclusion When no abnormal bands,the elevated HbA2 (>3.7% in this study)determined by Hb-CE could be used as a marker forβ-thalassaemia diagnosis,but theβ-thalassaemia co-existingα-thalassemia could not be differentiated fromβ-thalassaemia diagnosis.Decreased HbA2 level (<1.5% in this study)and HbH band could be used for the diagnosis of hemoglobin H disease.Only HbA2 determination by Hb-CE has no clinical sig-nificance for the screen and diagnosis ofα-thalassemia.

Objective To understand the pathogen of viral encephalitis(VE)in children,and establish a rapid and specific method for detecting human rhinovirus(HRV),and investigate the correlation between HRV and viral encephalitis(VE). Methods 169 CSF specimens were collected from children with convulsions and fever,who were admitted to the pediatric intensive care unit (PICU)of Second Affiliated Hospital of Shantou University Medical College between January 2012 and December 2012.Nested RT-PCR was used to detected HRV in CSF specimens,and the positive PCR products were se-quenced,then analyzed and constructed the phylogenetic tree by software.Results 39 (23.1%)out of 169 samples were HRV positive.Among them,148(87.6%)children were below 5 years old.The detection rate of HRV increases from July to September,and reached its highest point in September.Sequence analyzed showed that the 39 HRV positive specimens inclu-ding 18(46.1%,18/39)positive for HRV-A,7(17.9%,7/39)positive for HRV-B,14(35.9%,14/39)positive for HRV-C. There were 8 out of 28 VE cases were detected in HRV,including 3(50%,3/6)positive for HRV-C.Conclusion HRV could be detected in CSF specimens by nested RT-PCR,including three types of HRV,combined with clinical symptoms con-sidered that HRV may be one of the VE pathogen.

Objective To observe any rehabilitative effects of individualized exercise training in the treatment of rotator cuff injury in elite table tennis players.Methods Forty table tennis players from the Chinese National Team were studied.Twenty (the experimental group) had rotator cuff injuries and 20 without the injury formed the control group.An individualized rehabilitation treatment protocol was prepared for each of the players in the experimental group and implemented twice weekly for a total of 8 weeks.There was no intervention for the control group.Before and after the treatment,both groups were assessed using a questionnaire for disabilities of the arm,shoulder and hand (DASH),a motion assessment,and BIODEX isokinetic muscle testing.Results There were significant differences in average DASH scores between the two groups,with the experimental group scoring higher than the controls both before and after the treatment.Before the treatment,at angular velocities of 60°/s,180°/s and 300°/s the peak moments of the ER (external rotation,ER) and IR (internal rotation,IR) muscles and the ER/IR ratio were all lower in the experimental group than among the controls.However,after the treatment the performance of the experimental group had improved in all three tests.Conclusion The ER/IR ratio of the dominant shoulder of table tennis players with rotator cuff injury is lower than that of the players without the injury,but individualized rehabilitation treatment can effectively increase the ratio and help to improve their shoulder function.

Objective To investigate the mid-term effect of SolitaireAB stent-assisted interventional embolization with spring coils for the treatment of intracranial wide-necked aneurysms. Methods During the period from May 2009 to April 2013, a total of 49 patients with intracranial wide-necked aneurysm (49 aneurysms in total) received SolitaireAB stent-assisted interventional embolization treatment at authors’ hospital. In 41 patients, a total of 41 aneurysms were detected, of which ruptured aneurysm with bleeding was confirmed in 26 and un-ruptured aneurysm in 15. These 41 patients were followed up for 12-48 months. Based on modified Rankin scoring and DSA, CTA or MRA manifestations, the clinical results were evaluated. Results After the embolization treatment, re-bleeding of the aneurysm occurred in 2 cases, cerebral infarction in 3 cases, occlusion of the parent artery in one case and death in one case; the occurrence rate of complications was 14.2%. DSA, MRA or CTA performed at 12 months after the embolization treatment, showed that 32 aneurysms (78.0%) were completely obstructed, which was obviously higher than that observed on DSA performed immediately after the embolization procedure (21 aneurysms, 42.9%), the difference was statistically significant (P=0.02);residue of aneurismal neck was seen in 7 cases (17.1%) and partial occlusion in 2 cases (4.9%), which were much better than those observed on DSA that was performed immediately after the embolization procedure. Twenty-four aneurysms (58.5%) remained stable, showing no any change, and recurrence of aneurysm was observed in 4 cases (9.7%). At the last follow-up exam, the modified Rankin scoring showed that 0 point was seen in 18 cases (43.9%), one point in 10 cases (24.4%), 2 points in 5 cases (12.2%), 3 points in 4 cases (9.8%), 4 points in 2 cases (4.85%) and 5 points in 2 cases (4.85%). The self-care rate for daily activities was 80.5%, the prognosis was good. Conclusion For the treatment of intracranial wide-necked aneurysms, SolitaireAB stent-assisted interventional embolization with spring coils is safe and effective. This technique can improve the embolization rate and reduce the procedure-related complications.

Objective To investigate the relationship between Hepatitis B patients with different viral loads and immunoglob-ulin A,G,M and complement C3,C4.Methods Firstly,followed by real-time fluorescence quantitative PCR detection 210 cases of hepatitis B patients with HBV-DNA levels,according to 10n copies/ml different viral load detection results,it was divided into 102 ~108 copies/ml of the experimental groups.Then the experimental groups and control group were simulta-neously detected in immunoglobulin A,G,M and complement C3,C4.Analysed the correlation between HBV loads and im-munoglobulin A,G,M and complement C3,C4.Results When the viral loads of hepatitis B patients were 105 ~108 copies/ml,the testing results of IgA,IgG and IgM were both increasing (U =12.43,10.96,6.42,P <0.01),while C3 and C4 were both decreasing (U =8.37,6.0,P <0.01).When the viral loads of hepatitis B patients were 102 ~ 104 copies/ml,only IgA and IgM were increasing (U =2.36,2.04,P <0.05),the other testing results had no statistical significance.Between the test of 7 experimental groups compared with each other,only 104 group and 105 group had significantly changed (IgA and IgM were increasing,C4 was decreasing,U =2.39,2.46,2.09,P <0.05,IgG was increasing,U = 3.25,P <0.01),but between other low viral loads or high viral loads were not significantly differences.Conclusion The different viral loads of hepatitis B patients could cause the different changes of immunoglobulin A,G,M and complement C3,C4,especially in the 4 groups from 105 to 108 copies/ml.Followed by increasing in viral loads,there were immunoglobulin A,G,M increasing and comple-ment C3,C4 decreasing,and also serious impaction on the immune function of organism.There was a positive correlation be-tween viral loads in vivo and immune damages,correlation coefficient (γ =0.967,P <0.01).When the viral loads from 104 to 105 copies/ml,the testing results had changed significantly.It suggest that should control viral loads under 104 copies/ml in the hepatitis B antiviral treatment,so the effect of immune function damage will be the minimum.