Objective To diagnose the infection of Pseudomonas aeruginosa early. Methods Polymerase chain reaction (PCR) was used to amplify the fragment of Pseudomonas aeruginosa OprI gene. The fragment was determined by HaeⅢ and PvuⅡ digestion, and sequencing analyses. Results It showed that 96 of 223 specimens were cultured to be positive with Pseudomonas aeruginosa, 96 of which had expectant streaks. Otherwise the other specimens had no positive streaks. The procedure needed only 4 hours. The PCR products were determined by ribonuclease HaeⅢ and PvuⅡ , and resulted in two small fragments with 49bp and 112bp separately. By automatic sequencing analysis, the coincidence rate with the gene bank was 100%. Conclusions The results indicates that the OprI gene detection by PCR is a specific, sensitive and quick technique for the diagnosis of Pseudomonas aeruginosa infection.

ObjectiveTo investigate the changes of hepatitis B sarfaceantigen (HBsAg) titer in hepatitis B e antigen (HBeAg) positive chronic hepatitis B (CHB) patients treated with nucleos(t)ide analogs and pegylated interferon alfa-2a (PEG IFNα-2a) sequential therapy.Methods Among 6 HBeAg positive CHB patients,3 patients were treated with nucleos(t)ide analogs followed by PEG IFNα-2a for 48 weeks,3 patients were treated with nucleos(t)ide analog monotherapy.The serum HBsAg,anti-HBs,HBeAg,anti-HBe andanti-HBcweredetectedusingthetime-resolved immunofluorometric assay and serum hepatitis B virus (HBV) DNA levels were determined by Taqman polymerase chain reaction (PCR) every 12 weeks.Results HBsAg loss were achieved in three patients after 48-week nucleos(t)ide analogs and PEG IFNα-2a sequential therapy.However,the HBsAg titers of another 3 patients varied from 100 IU/mL to 320 IU/mL.ConclusionIn HBeAg positive CHB patients who obtain virologic response accompanied with HBsAg titer decreasing dramatically by nucleos(t) ide analog treatment,PEG IFNα-2a sequential treatment can increase HBsAg clearance rate.

Objective To explore the influence of treatment duration on the prognosis of Cryptococcus neoformans meningitis (CNM). Methods The clinical data of 30 patients with CNM were retrospectively analyzed. The time of Cryptococcus neoformans culture in cerebrospinal fluid turned to be negative were compared between patients with and without amphotericin B (AmB)endoneurilemma injection to determine whether endoneurilemma injection would make any difference on prognosis of CNM. Statistic analysis was performed by t test. Results The time of Cryptococcus neoformans culture in cerebrospinal fluid turned to be negative was (22. 73 ± 10. 00) days in 11patients with AmB endoneurilemma injection compared to (18. 92±7. 26) days in 13 patients without endoneurilemma injection. The difference was not statistically significant (t = 0. 172,P>0. 05). The effective rate was 45. 8% after 2-week anti Cryptococcus neoformans treatment, which increased to 100.0% after 6-week treatment. However, the curative rate at week 6 of treatment was 0. All ten patients who finished the treatment (three to four months' treatment) were acquired effective.Conclusion Treatment with standard dosage and adequate duration is crucial in improving the prognosis of Cryptococcus neoformans meningitis.

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Objective To provide the guidance for the control and treatment of blood stream infec-tion caused by Serratia marcescens strains through analyzing the homology and drug resistant genes of the iso-lates collected from the Intensive Care Unit ( ICU) of Shaoxing County Central Hospital.Methods Serratia marcescens strains were isolated from ICU patients with blood stream infection and also from the hands of health care providers in the ICU from June 1st to September 30th, 2013.The antibiotic susceptibilities of the Serratia marcescens isolates were tested.PCR was performed to amplify the common drug resistant genes. Pulse-field gel electrophoresis ( PFGE) was carried out for analyzing the homology of all isolates.The com-plete clinical data of the patients were collected and statistically analyzed with Spearman′s rank correlation coefficient.Results Seventeen strains were isolated in this study.All of the 17 strains were resistant to the first and second generation Cephalosporin, Gentamicin and Ciprofloxacin, and sensitive to Amikacin and Ceftazidime.The drug resistant rates to Carbapenems ranged from 11.76%to 35.29%.One of the isolates (5.88%) carried the TEM gene.The results of PFGE showed that the phenotypes of all isolates were identi-cal.Conclusion Serratia marcescens strains were critical hospital infectious pathogens.They were able to spread in the hospital and were resistant to multiple antibiotics.Clinical physicians should properly use anti-biotics for the patients based on the result of drug susceptibility test.A control regulation for Serratia marces-cens infection within hospital should be enforced to avoid the cross infection and the outbreak of resistant strains.

Objective To investigate the efficacy and safety of adefovir dipivoxil (ADV) in treatment of chronic hepatitis B (CHB) patients with lamivudine (LAM) resistance. Methods There were treatment group (32 CHB patients with LAM resistance) and historical control group (24 CHB patients with LAM resistance) in this study. The treatment group received ADV 10 mg/d and LAM 100 mg/d for 48 weeks; the historical control group continued to use LAM monotherapy. During the treatment causes, serum HBV DNA levels, liver function and HBV serology were monitored regularly, and safety assessments were also conducted. Results In treatment group, mean HBV DNA levels decreased by 2.56 log10 eopies/ml and 2.93 log10 copies/ml, virus response rates were 50. 0% and 75.0%, ALT normalization rates were 53.1% and 68.8% after 24 and 48 weeks of treatment, respectively. The histological improvement rate was 65.6% after 48 weeks. Comparing with those in control group, the differences were statistically significant ( P <0. 05), while there was no significant statistical differences in HBeAg loss rate and HBeAg seroconversion rate between two groups. There was no severe adverse event during the treatment. Conclusion ADV is effective and safe in treatment of lamivudine-resistant CHB.

Objective To study the dynamics changes of cerebrospinal fluid (CSF) bacterial load within 48 h after infection in a rabbit meningitis model, and provide information for diagnosis,treatment and prognosis of this disease. Methods Taking New Zealand white rabbit as the study object, meningitis model was established via cerebellar cistern puncture with different concentrations of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) to explore the relationship between the mortality of animals and the subarachnoid inoculation dosage. The dynamics study of CSF bacterial load was conducted with proper inoculation bacterial dosage. Forty-eight rabbits were separated into four groups (12 each group): E. coli meningitis model group, E. coli meningitis + ceftriaxone treated group, S. aureus meningitis model group and S. aureus meningitis + vancomycin treated group. At 0,12, 24, 36 and 48 h of inoculation, CSF and blood samples were obtained for CSF bacterial quantitative culture, CSF leukocyte count and peripheral blood leukocyte count. Finally, the relationships between the early mortality of animals, the efficacy of antibiotics, CSF leukocyte counts and the dynamics changes of CSF bacterial load were analyzed in the bacterial meningitis rabbit model.The CSF bacterial load and the white blood cell count curve were compared by analysis of covariance (ANOVA). Correlation test was done using correlate partial analysis. Results The relationship between subarachnoid inoculation dosage and the mortality of rabbits presented S-curve correlation.The bacterial load in subarachnoid space peaked in 12-24 h after infection and then gradually decreased. Effective antibiotic therapy could significantly speed up the decline of this process. There were significantly different between E. coli meningitis model group and E. coli meningitis+ceftriaxone treated group (F= 27. 10, P<0. 01), between S. aureus meningitis model group and S. aureus meningitis + vancomycin treated group (F=5. 97, P = 0. 016). There was a positive correlation between CSF bacterial load and CSF leukocyte count in E. coli and S. aureus meningitis model groups (r=0. 89, 0.84, respectively; P = 0.046, 0.049, respectively). Conclusions In the treatment of bacterial meningitis, effective and sufficient antibiotics should be used as soon as possible to control the CSF bacterial load and reduce the mortality. The CSF leukocyte count can be used as indicator of CSF bacterial load and guide the antibiotic treatment in clinical bacterial meningitis.

BACKGROUND: Carbonated hydroxyapatite cement (CHC) s a new kind of biomaterial for bone defect, which is made of powder and fluid, and can be mixed to be pasty to repair various bone defects.OBJECTIVE: To observe the improvement of vertebrae height and pain in patients with osteoporosis vertebral compression fracture (VCF) after vertebroplasty by using a new kind of bone graft biomaterial, taking CHC as the filling material to reinforce the vertebral body.DESIGN: A contrast observation trial taking patients as subjects.SETTING: Department of Orthopaedics, General Hospital of Chinese PLA.PARTICIPANTS: Totally 34 patients with thoracic or lumbar osteoporosis VCF who received the treatment in the Department of Orthopaedics, General Hospital of Chinese PLA between October 2000 and August 2003. Inclusive criteria: ①Definite diagnosis by CT; ② Informed consents were obtained from the patients. Exclusive criteria: The patients with osteoporosis vertebral compression fractures who suffered vertebral posterior wall fracture. There were 6 males and 28 females, and they were aged (72±13)years; Among the patients, 27 were diagnosed as postmenopausal osteoporosis, 1 as cortical hormone-induced osteoporosis and 6 male patients weresenile osteoporosis.METHODS: ①All the patients were randomly divided into two groups: Experimental group (n =23) and control group (n=11). All the patients were performed percutaneous operation with local anesthenia. All cases were performed percutaneous operation under local anesthesia. Under the C-arm monitored, one side pedicle puncture was performed to enter the anterior column of the involved VCF. Patients of the experimental group were filled with CHC. Patients of control group were filled with polymethyl Methacrylate (PMMA) with the same way. ② Referred to McGill-Melzack scoring. Among the scale 0-100 mm (0 was no pain, 100 was acute pain), the value indicated the painful intensity and mental assault degree. ＜ 30 scores indicated good, 30-40 basically satisfied and ≥ 50 poor .③ Referred to the method from Lee et al, the preoperative height (A1) and postoperative height (A2) of compression fracture position of VCF were measured according to the lateral X-ray film. At the same time, the upper vertebral height (A3) and the inferior vertebral height (A4) were measured at the same position. The original height (A) of the involved vertebra was calculated as (A)= (A3+A4)/2,and the preoperative vertebral compression rate =(A-A1 )/A, the postoperative vertebral compression rate =(A-A2)/A, the restoring rate = (the preoperative vertebral compression rate-the postoperative vertebral compression rate)/the preoperative vertebral compression rate. ④ The wounds of the patients were observed after operation. The levels of blood routine, serum calcium and serum phosphorus were detected before, one day and one week after operation. MAIN OUTCOME MEASURES: ① Preoperative and postoperative VAS scoring. ② The vertebral compression rate and restoring rate. ③ Wounds were observed after operation. The blood routine, the serum calcium and serum phosphorus were detected before, one day and one week after operation.RESULTS: Totally 34 patients were involved in the result analysis. ①The preoperative visual analogue scale (VAS) score of experimental group were (91.5±21.7) points, and the postoperative ones were (44.5±27.2) points. The difference of VAS score reduced gradually along with the postoperative time. There was no difference of VAS score between experimental group and the control group 4 weeks after operation. ② The biocompatibility of CHC in the vertebral body was fine. The vertebral compression rate of experimental group was recovered from (43.1±21.4)％ preoperatively to (27.3± 18.5)％ postoperatively. The rate of restored heights was (27.3±18.5)％. ③ All patients obtained Ⅰ stage wound healing, and none of them had infection, inflammatory secretion and nervous symptom. There were no differences in blood routine test, serum calcium, serum phosphorus between patients in two groups. One case filled by PMMA and two cases filled by CHC presented leakage, and none had nervous symptom.CONCLUSION: As the filling materials for vertebropalsty, CHC can restore the vertebral heights and relieve pain safely and effectively, however, its efficacy to relieve pain is not significant as PMMA in the short term.

Objective To establish normally immortalized porcine hepatocyte lines by ectopic expression of simian virus 40 large T (SV40LT) antigen and the human telomerase reverse transcriptase(hTERT). Methods Primary porcine Hepatoeyte cells were transfeeted with recombinant retrovirus containing SV40LT or hTERT respectively. Subsequently drug resistant cell clones were screened and expanded for further studies. Immortalized porcine hepatocyte was confirmed by examination. Results The morphological phenotype of the transfected cells was similar to the primary porcine hepatocyte. One clone, HepLP, has been maintained in cultue for half year, and expanded by more than 60 passages. SV40 LT and hTERT could be detected in transfected porcine hepatocyte. Pig albumin mRNA was also detected by RT-PCR. No tumor formation occurred when HepLP cells were injected into Balb/c nude mice. Conclusions The immortalized, nontumorigenic, porcine hepatoeytes maintained the properties of porcine primary hepatocytes such as the albumin secretion. This generation of immortalized porcine hepatocyte may be helpful for bioartifical liver support system, hepatocytes transplantation, drug/toxicological studies, and liver biologic studies.