In this review, the analytical methods of polyethylene glycol-modified proteins are introduced, including spectroscopy, liquid chromatography, electrophoresis, mass spectrometry, etc. These methods are compared in respect to their individual characteristics.

Objective To observe the effect of moxibustion on the proliferation of neural stem cells after cerebral ischemia- reperfusion in rat models.Method The middle cerebral artery occlusion (MCAO) model was developed by using modified Zea-Longa method. Seventy-two Wistar rats were randomized into group A (sham-operation group), group B (model group) and group C (moxibustion group). Each group was then further divided into 4 sub-groups according to different time points: 3 d, 7 d, 14 d, and 21 d, 6 rats in each sub-group. For each group, Baihui (GV 20) and bilateral Taiyang (EX-HN 5) were selected to study. The proliferated cells were marked by intraperitoneal injection of BrdU, and TTC staining and HE staining were adopted to detect the pathological changes of brain tissues after cerebral ischemia-reperfusion. Immunohistochemical method was used to dynamically detect the positive expressions of BrdU and Nestin cells in DG and SVZ zones at different time points, and microscope image analysis was taken to study the positive expressions.Result The neurological deficit score in group C was significantly lower than that in group B at each time point (P<0.05). The neurological deficit score in group C was significantly different from that in group B at the same time point (P<0.05). Moxibustion can up-regulate the expression of BrdU in SVZ and DG, and the increase started from the 3rd day and reached the peak on the 7th day. The increase of BrdU expression in group C was more significant than that in group B (P<0.05). Moxibustion can up-regulate the expression of Nestin in SVZ and DG, and the increase started from the 3rd day and reached the peak on the 7th day. The increase of Nestin expression in group C was more significant than that in group B (P<0.05).Conclusion Moxibustion can remarkably promote the recovery of nerve function and the proliferation of BrdU and Nestin after cerebral ischemia-reperfusion in rats.

Objective To evaluate the inhibitory effect of phase change doxorubicin loaded carboxymethyl hexanoyl chitosan nanodroplet on ovarian cancer cells,and the effect of its ultrasound image in vitro.Methods The carboxymethyl hexanoyl chitosan synthesized through the acylationreaction with carboxymethyl chitosan and hexanoic anhydride.The drug loaded carboxymethyl hexanoyl chitosan nanodroplets were prepared by ultrasonic emulsification.The surface morphology,particle diameter and electric potential were characterized.Ultrasound imaging of the nanodroplet was evaluated in vitro.The encapsulation efficiency was determined by ultraviolet spectrophotometry.The survival rate of ovarian cancer cell was detected using CCK-8 reagent.The statistical analysis was performed.Results The drug loaded carboxymethyl hexanoyl chitosan nanodroplet was successfully prepared which showed regular morphology in microscope,the mean diameter of (458.33± 43.50)nm.The encapsulation efficiency was (52.06 ± 10.14)％.The nanodroplet could enhance ultrasonic imaging.The survival rate of ultrasound combined with drug loaded nanodroplet group ([62.54± 3.60]％) was lower than those of the free drug group ([75.55±7.21]％) and drug loaded nanodroplet group ([76.18±4.94]％),ultrasound group ([89.90±0.83]％;P＜0.05).Conclusion Ultrasound-mediated drug loaded nanodroplet can inhibit ovarian cancer cells,and has the potential for application in the clinical diagnosis and treatment.

To provide the basis for preparation of diagnostic kits and vaccines in Toxoplasma gondii infection, the gene coding for the qualified recombinant p30 protein (SAG1) of this parasite was amplified by PCR, and the amplified gene was cloned into prokaryotic expression vector pET-30a(+) to construct the recombinant plasmid, and then transformed to E.coli DH5α. The positive recombinant plasmid was screened by PCR and double enzymes digestion, and the nucleotide sequence of p30 gene was determined by automated DNA sequencing. Meanwhile, the identified recombinant plasmid was transformed to E.coli BL21(DE3) with the expression of p30 on bacteria induced by IPTG and the expressed protein was identified by SDS-PAGE. The protein obtained was then further purified and refolded, and its biological activity was checked by Western blotting. It was shown that the size of the amplified gene was 750 bp with molecular weight of 30 ku, and this protein could specifically react with monoclonal antibody against p30 protein.

Objective To discuss the effect of team-based learning(TBL)combined with net-work environment in pathological experiment teaching. Methods Totally 112 clinical medical under-graduates were selected as research object. Digital microscope interactive teaching platform under the network environment was used in experimental group(n=56). In pathology experiment teaching,TBL steps of training,teaching plan demonstrating,grouping,teaching task arranging,asking questions, group learning and communicating were implemented. Traditional method (lecture)was used in con-trol group (n=56). Teaching effect was analyzed through questionnaire survey and score analysis. Stu-dents' evaluation on TBL teaching and network environment resources was expressed as percentage. Results In experiment group,average score was (85.5±3.11)points and average final exam score was (83.8±2.53)points,while in control group,average final exam score was (76.4±11.89)points. There were significance differences between the two groups (t=7.018,P<0.01). Surveys showed that most students accepted TBL teaching combined with network environment. Conclusions TBL teach-ing combined with network environment is feasible and effective in the pathology experimental class. Students' learning enthusiasm is generally improved and effectiveness of TBL teaching is satisfactory.

Objective To discuss the presentations, pathologic features, diagnosis and treatment of primary synovial sarcoma of the kidney. Methods One case of primary synovial sarcoma of the kidney was reported and the relevant literature was reviewed. A 55-year-old man was admitted with complaint of right abdomen and flank pain for 5 h. Computerized tomography revealed a 12.5 cm × 11.0 cm × 9. 0 cm mass located at the middle and lower pole of the right kidney. The patient was taken radical nephrectomy. Results The diagnosis of primary synovial sarcoma of the kidney in the patient was confirmed by postoperative pathology. Under microscope, tumor was typically mitotically active, monomorphic spindle cells growing in intersecting fascicles or in solid sheets with epithelial differentiation. In some areas a haemangiopericytoma-like pattern was found. Immunohistochemical staining showed that the tumor cells were positive for the markers Vimentin, CD99 and Bcl-2, but CK was negative. The patient died of local recurrence and multi-metastasis at 8 months after surgery. Conclusions Primary synovial sarcoma of the kidney is extremely rare with a high grade of malignancy,and its prognosis is poor. The diagnosis depends on pathological features, Immunohistochemical studies and RT-PCR detection. Radical resection combined with chemicaltherapy is considered to be the most reliable treatment so far.

Objective To propose SHA.LIN nephrolithometry scoring system for assessing and predicting the stone-free rate of percutaneous nephrolithotomy ( PCNL) and to investigate the clinical value of SHA.LIN scoring system for nephrolithiasis in patients undergoing PCNL .Methods A literature review from 1976 to 2014 was performed to identify clinically relevant and reproducible variables that could affect the outcomes of PCNL. Six reproducible variables available from preoperative noncontrast-enhanced computed tomography were measured , including stone size ( S) , hydronephrosis ( H) , anatomic distribution (A), length of tract(L), indicator of CT(I), number of involved calices(N) and was named as SHA.LIN nephrolithometry scoring system .A retrospective analysis was conducted of clinical data of 116 patients with nephrolithiasis undergoing PCNL from June 2011 to March 2015. The general conditions , preoperative information , stone characteristics and perioperative variables were collected . The correlation of nephrolithometry scores based on SHA.LIN scoring system with stone-free status, operation time, blood loss, length of hospital stay and postoperative complications were analyzed . Receiver operating characteristic ( ROC) curves was drawn to detect sensitivity and specificity of SHA .LIN score in predicting the stone-free rates of PCNL.Results The SHA.LIN score was 9.13 ±2.24 in this cohort.The stone free rate was 75.9%(88/116).Postoperative complications occurred in 32 (27.6%) cases.In those patients with stone
free, the SHA.LIN score was 8.27 ±1.62, significantly lower than that in those patients with residual stones 11.86 ±1.72 ( t =-10.069, P=0.000) .The SHA.LIN score showed significant correlation with the postoperative stone free status, operation time, estimated blood loss (P<0.01).But, it did not correlate with postoperative complications and length of hospital stay (P>0.05).The area under curve of ROC curves for the SHA.LIN scoring system was 0.923 ( 95%CI 0.870 -0.975 ) . Conclusions The SHA.LIN nephrolithometry scoring system can predict postoperative stone-free status of PCNL and can be used for disease related assessment.Further research is required to evaluate its performance in predicting peri-operative variables and postoperative complications .

This paper reports one case of atypical falciparum malaria imported from Africa,whose blood smear contains many large trophozoites,with punctiform or massive brown pigment granules,the body shape of the plasmodium is similar to that of Plas-modium vivax and Plasmodium ovale. After the gene detection by PCR,the case was diagnosed as falciparum malaria. As large tro-phozoites were rarely seen in the peripheral blood of non-severe falciparum malaria cases,much attention should be paid to the identification of Plasmodium falciparum and other plasmodia in microscopic examinations.

OBJECTIVETo probe the significance of specific IgG4 in sera of patients with cerebral cysticercosis for diagnosis and therapeutic evaluation.

METHODSSpecific IgG4 in sera of patients with cerebral cysticercosis was assessed using colloidal gold-labeled mouse-anti-human IgG4 McAb as probe. The results were compared with the CT image manifestation.

RESULTSThe specific IgG4 positive rate in sera of patients with cerebral cysticercosis was 97.8%, whereas sera from patients with other kinds of parasitosis or central nerve system disease and the control group were all negative, except for a weak cross-reaction of sera from patients with hepatic echinococoosis. The determination of specific IgG4 in sera of patients with cerebral cysticercosis during different times of treatment showed that along with an increase in treatment time and improvement of clinical symptoms, specific IgG4 level gradually decreased. The positive rate and intensity of specific IgG4 in sera from patients with cerebral cysticercosis were consistent with the number of cysticercus parasites in the brain and pathologic changes, such as survival, disintegration, death and calcification. Survival of cysticercus in the brain was objectively evaluated using this technique.

CONCLUSIONSThe determination of specific IgG4 in sera is a practical method for diagnosis and therapeutic evaluation of cerebral cysticercosis.

Animals ; Antibodies, Monoclonal ; immunology ; Antibody Specificity ; Humans ; Immunoglobulin G ; blood ; immunology ; Mice ; Mice, Inbred BALB C ; Neurocysticercosis ; blood ; diagnosis ; therapy ; Predictive Value of Tests ; Tomography, X-Ray Computed

Objective To analyze the genotypes and homology of MSP?1 and CSP gene of Plasmodium vivax in Shandong Province,so as to provide the evidence for case traceability. Methods A total of 12 blood samples were collected from P. vivax?infected cases in Shandong Province in 2011. Parasite genomic DNA was extracted. Primers were designed according to MSP?1 and CSP gene sequences of P. vivax. Then Nested PCR,enzyme digestion,sequencing and sequence alignment,and homolo?gous analysis were performed. Results The MSP?1 gene of all the 12 samples from P. vivax?infected cases were detected with a 470 bp PCR amplification band,and 350 bp and 120 bp enzyme digestion fragments,which were identified as type Sal?1. An analysis of phylogenetic tree of MSP?1 gene showed that the sequences of 9 indigenous case samples in Shandong Province were located in the same branch,one case sample infected from India was located in the same branch with India strains. All the 12 P. vivax?infected samples covered GDRA(D/A)GQPA sequences in CSP gene,which were identified as type PV?Ⅰ. Of the CSP gene among 12 P. vivax?infected samples,10 samples of indigenous case in Shandong Province and one sample of the case in?fected in Guangdong Province were detected with both 560-840 bp and 150-230 bp PCR amplification bands,which were iden?tified as temperate zone family strain of type PV?Ⅰ. However,one sample from the case infected in India was detected only with a 560-840 bp band,which was identified as tropical zone family strain of PV?Ⅰ. An analysis of phylogenetic tree of CSP gene showed that the sequences of 10 samples from the indigenous cases in Shandong Province and one sample from the case infected in Guangdong Province were located in the same branch,one sample from the case infected in India was located in the same branch with India and Indonesia strains. Conclusion Of all the indigenous isolates in Shandong Province,MSP?1 gene is geno?typed type Sal?1,CSP gene is genotyped temperate zone family strain of type PV?Ⅰ,with a high homology found among the in?digenous isolates.