Objective To investigate the curative effect of pidotimod combined with Yanhuning for children with refractory mycoplasma pneumoniae pneumonia(RMPP).Methods 150 patients with RMPP in our hospital were divided into 3 groups,including Yanhuning group(A group),azithromycin combined with Yanhuning group(B group) and pidotimod combined with Yanhuning group(C group).And the total effective rate,the clinical symptoms,immune function parameters were analyzed.Results Compared to A group,the effective rate of B group and C group were sig-nificantly higher(all P <0.05),but there was no statistically significant difference between B group and C group. Compared to A group,the defervescence time,cough disappeared time and rales disappeared time were significantly shorter(all P <0.05),and the effect in C group was better than that in B group (all P <0.05).Compared to normal control group,CD +3 ,CD +4 ,CD +4 /CD +8 and CD +16 CD +56 in A group,B group and C group before the treatment were sig-nificantly lower(P <0.05),but IgG,IgM,CD +8 and CD +19 were significantly higher(all P <0.05).And CD +3 ,CD +4 , CD +4 /CD +8 and CD +16C D +56 in C group were significantly higher(all P <0.05)after the treatment.During follow -up, compared to A group,the recurrence rate and time in B group and C group were significant reduction(all P <0.05), and the recurrence rate and time in C group were less than that in B group (all P <0.05).So the regulation of immune function in C group was more significant than that in A group and B group,the prognosis was better,and the repeated times and continuous time were decreased.Conclusion Pidotimod combined with Yanhuning treatment of RMPP can regulate the immune function of children,improve clinical symptoms,which should be promoted.

ObjectiveFor finding out the early stage growth circumstance of high risk neonate, we studied continuously recently years through regular checkups and early stage education directing. Methods1040 high risk neonates were followed up and checked once 1-3 months.ResultsThey are 2 cases cerebral palsy and 15 cases sports growth backward with posture abnormal. Through early stage restored training and treating, all of them are improved on the sports growth and posture. ConclusionsEarly intervention, regular checkups and early stage education, is a useful way for high risk neonate.

BACKGROUND:Because of the special and complicated biological characteristics of oocytes, the suitable cyropreservation technology for oocytes faces more various chalenges. However, the uneven survival rate and fertilization, damages of developmental potential of the thawed oocyte stil exist. OBJECTIVE:To introduce the progress in basic and application researches of oocyte cryopreservation technology, and to iluminate the technical defects and thoughts and possible research points to overcome these problems. METHODS: A computer-based search of CNKI and PubMed was performed for articles concerning oocyte cryopreservation from January 2004 to October 2014. The search terms were oocyte, cryopreservation, vitrification in Chinese and English, respectively. The old and repeated articles were excluded. Finaly, 41 articles were included in result analysis. RESULTS AND CONCLUSION:The cryopreservation of oocytes by slow freezing and vitrification has been used in clinic. At the same time, the uneven survival and fertilization rate, damaged developmental potential of the thawed oocyte stil puzzle the clinicians. The key point to breakthrough or improvement of oocyte cryopreservation technology is the systematization of the protocol for oocylte cryopreservation, for example, the choice of cryoprotective agents, the development of carriers for oocytes, and the determination of oocyte stage. Furthermore, other related technologies should also be considered, including thein vitro mature technology of oocytes, cryopreservation and transplantation of ovary tissues.

BACKGROUND:The key of vertebroplasty and percutaneous kyphoplasty to success is whether the puncture needle can accurately reach the vertebral body through pedicle. Therefore, it is important to identify the correct point and direction of needling in the X-ray fluoroscopy. Among many methods published in present reports, the puncture point and the puncture angle are not fixed. Few reports concerned whether the puncture needle perforated pedicle medial wal . OBJECTIVE:To seek safe, effective puncture point and the puncture angle of percutaneous pedicle from the perspective of anatomy and radiography. METHODS:The best entry point during percutaneous vertebroplasty in the X-ray fluoroscopy:dissection was performed on thoracic, lumbar skeletal samples (T 6-L 5 ) to find the position of pedicle axis leading to the rear of the vertebral body, and this position is the best entry point of percutaneous vertebroplasty. It was fixed with mini-screw. The relationship of the best entry point and pedicle developing position in the X-ray fluoroscopy was analyzed to find the best entry point in the X-ray fluoroscopy. The best entry angle during percutaneous vertebroplasty:The average included angle of pedicle axis and vertebral sagittal line was measured using autopsy and CT scanning on adult thoracic and lumbar skeletal samples (T 6-L 5 ). The best entry angle during percutaneous vertebroplasty was found. RESULTS AND CONCLUSION:During percutaneous vertebroplasty, the best entry point in the X-ray fluoroscopy was the left pedicle projection 9 area and right pedicle projection 3 area. The optimal needle angle during percutaneous vertebroplasty:5°-10° in lumbar vertebra L1-L4;20° in L5, not more than 25°;about 5° in thoracic vertebra T6-T12 .

Objective To investigate antimicrobial prophylaxis in clean operation,and provide evidence for formu-lating antimicrobial management measures.Methods 12 types of clean operation cases who discharged from a tertia-ry first-class hospital in May of 2011 -2014 were selected,120 cases were selected each year,patients were divided into group I and group II according to the types of operation,survey forms were designed,assessment criteria for the rationality of antimicrobial use was formulated, antimicrobial use in different years was compared. Results Prophylactic use of antimicrobial agents for clean operation decreased from 93.33% in 2011 to 35.00% in 2013 and 2014;rates of correct medication time increased from 43.75% in 2011 to 97.62% in 2013 and 92.86% in 2014 respectively;rates of medication according to indications,rational choice of antimicrobial agents,and rational treatment course increased from 42.86%,60.71 %,and 21 .43% in 2011 to 100.00% in 2014 (all P < 0.05 ). Conclusion Special rectification activities of antimicrobial use effectively promoted the standard application of anti-microbial agents for clean operation.

Objective Up to now, there has been no sure cure for genital herpes (GH), and vaccine seems a most promis-ing approach to the prevention and treatment of herpes simplex virus Ⅱ(HSV-2) infection.In this study, we investigated the feasibili-ty of preparing a dendritic cell ( DC) vaccine modified by the adenovirus-mediated HSV-2 gD gene. Methods We subcloned the HSV-2 gD gene into the vector Shuttle-2 and constructed the recombinant adenovirus pAdeno-HSV-2 gD following identification by en-zyme digestion and DNA sequence analysis .We isolated DCs from the mouse bone marrow , analyzed their phenotypes by flow cytome-try after transfection with the recombinant adenovirus pAdeno-HSV-2 gD, and determined the expression of HSV-2 gD by immunohisto-chemistry, RT-PCR, SDS-PAGE, and Western blot. Results Based on HSV-2 DNA, the corresponding target fragments were am-plified with the gD gene primers.Agarose gel electrophoresis showed the correct size of the PCR product (1182 bp) as predicted.The recombinant adenovirus pAdeno-HSV-2 gD was obtained by transfecting the 293 cells with pAdeno-gD DNA, which had an activity of 4 ×1010 IU/mL.The contents of CD40, CD80, and CD86 were (74.2 ±3.9), (73.9 ±4.1), and (76.1 ±5.5) % in the mature DCs and (81.3 ±3.1), (80.4 ±2.9), and (83.7 ±3.9) % in the pAdeno-HSV-2 gD DCs, significantly increased as compared with those in the immature DCs ([9.7 ±0.5], [7.5 ±1.2], and [5.2 ±1.1] %) (P<0.01).No statistically significant differ-ences were found between the expression of the surface molecules in the pAdeno -HSV-2 gD DCs and that in the cytokine stimulation-in-duced mature DCs (P>0.05).RT-PCR and immunohistochemistry confirmed the expression of HSV-2 gD in DCs.SDS-PAGE and Western blot of the expressed protein showed a new band with an apparent molecular mass corresponding to the predicted size (43000). Conclusion The results of our study have paved the ground for the successful preparation and identification of a dendritic cell vaccine modified by the adenovirus-mediated HSV-2 gD gene.

Objective To evaluate the specific immune response induced by a dendritic cell-based adenovirus-mediated vaccine carrying the herpes simplex virus type 2 glycoprotein D gene (pAdeno-HSV-2 gD-DC) in BALB/c mice.Methods Forty BALB/c mice were equally divided into four groups:blank control group receiving no treatment,pAdeno-DC group immunized with pAdeno-DC,pAdeno-HSV-2 gD-DC group immunized with the previously constructed vaccine pAdeno-HSV-2 gD-DC,DC group immunized with DCs only.Totally,three rounds of vaccination were conducted at a 7-day interval.Ten days after the last vaccination,serum samples were collected and spleen cells were isolated from these mice.Enzyme-linked immunosorbent assay (ELISA) was performed to measure the level of IgG antibody against HSV-2 gD in the serum samples.Some spleen cells were stimulated with HSV-2 gD protein (10 mg/L) for 72 hours; then,ELISA was carried out to determine the levels of interferon (IFN)-γand interleukin (IL)-4 in the supernatant,and 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay to estimate the proliferative activity of these cells.The cytotoxicity of spleen cells was also evaluated based on the measurement of lactate dehydrogenase (LDH) release.Results The serum level of IgG antibody against HSV-2 gD (given in the absorbance value at 450 nm) was 0.313 ± 0.034 in the pAdeno-HSV-2 gD-DC group,significantly higher than that in the pAdeno-DC group,DC group and blank control group (0.034 ± 0.009,0.028 ± 0.009 and 0.026 ± 0.010 respectively,all P ＜ 0.05).Increased proliferative activity and cytotoxicity were observed in spleen cells from the pAdeno-HSV-2 gD-DC group compared with those from the pAdeno-DC group,DC group and blank control group (cell stimulation index:1.600 ± 0.215 vs.1.063 ± 0.070,1.056 ± 0.063 and 1.020 ± 0.051,all P ＜ 0.05; percentage of cytotoxicity:37.1％ vs.16.0％,14.9％ and 15.7％,all P ＜ 0.05).The levels of IFN-γ and IL-4 (both given in the absorbance value at 450 nm) were 0.568 ± 0.031 and 0.544-± 0.043 respectively in the supernatant of spleen cells from the pAdeno-HSV-2 gD-DC group,compared to 0.266 ± 0.021 and 0.278 ± 0.037 respectively in the pAdeno-DC group (bothP＜ 0.05),0.271 ± 0.023 and 0.275 ± 0.044 respectively in the DC group (bothP＜ 0.05),and 0.252 ± 0.012 and 0.245 ± 0.051 respectively in the blank control group (both P＜ 0.05).Conclusion The vaccine pAdenoHSV-2 gD-DC could induce a specific and strong immune response in BALB/c mice.

Objective To evaluate the efficacy and safety of linezolid in empirical treatment of methicillin-resistant Staphylococcus (MRS) pneumonia. Methods One hundred and thirty-five hospitalized patients with MSR pneumonia receiving linezolid from April 2009 to October 2010 were enrolled in this retrospective cohort study, and all subjects were assigned to two groups: 75 cases with empirical treatment (linezolid 0. 6 g by infusion q12h at admission) , and 60 cases with objective treatment (linezolid after the sputum culture). The severity score, clinical effect and adverse effect were observed, and the therapeutic effects in patients with high risk factors were especially evaluated. SPSS13.0 software was used for statistical analysis. Results The scores were decreased significantly after finishing therapeutic causes for 3 and 7 days in both groups (tempirical =12.29 and 16.53, tobjective =9.36 and 11.49, P<0. 05). There were significant differences in severity scores after 3 and 7 days between empirical and objective treatment groups (t =2. 64 and 3. 08, P < 0. 01). There were significant differences in absorption time, length of ICU and total hospital stay between two groups (t =6. 61 , 4. 39 and 10. 25, P <0. 05). In empirical and objective group, the effective rates were 88.0% (66/75) and 83.3% (50/60) (x2 = 0.60, P > 0.05 ). In the patients with high risk factors, the effective rates of two groups were 86. 8% (33/38) and 63. 6% (14/22) , and the difference was significant (x2 =4.42, P<0.05). The rate of adverse effects were 6.7% (5/75) in empirical group and 5.0% (3/60) in objective group, and the difference was not significant (x2 =0. 17, P > 0. 05). Conclusion Linezolid can be used as empirical treatment for MRS pneumonia with rapid symptoms relieve and high efficacy, especially for patients with high risk.