According to that Y-chromosomal sequence in characterised by Y-specific repeat DNA family (DYZ1 ), which contain 800-5000 copies, a pair of primers Y3, Y4 is designed to amplify their 446bp long of specific DNA segment by polymerase chain reaction (PCR), so as to detect male fetal cells in materal blood. In this paper male fetal cells in maternal blood can be detected by PCR amplification of unpurified DNA from maternal peripheral blood during various stages of gestation (early, middle, late). Compared with villi, ammotic fluid and deliverd neonate sex their coincident rate are 93%. 100%, 87. 5% respectively among three periods. It is revealed that noninvasive examining fetal cells from peripheral blood of pregnant women for diagnosis of sex-linked inherited diseases is significant valuable.

According to ZFY gene sequence on Y-chromosome, which supposedly codes for the testis determining factor (TDF), a pair of primers F3.2 and F4.2 were designed to specifically amplify 650 bp DNA fragment. The amplified product was electrophoresized on 1.5% agarose gel stained with EB, and then directly visualized under UV. This protocol resulted in constantly producing 650 bp band of the specific product by amplification of 500 pg male genomic DNA and had the same result when mixed with 100 fold amount of female genomic DNA. When this method was used to detect Y-ZFY of chorionic villi and amniotic fluid in 20 samples respectively, its accuracy for determining fetal sex reached 100 percent.

Objective To evaluate the incidence of autoimmune thyroid disease (AITD) in systemic lupus erythematosus (SLE) and examine the correlation between AITD and SLE activity.Methods The study group included 220 SLE patients with the screening of thyroid function (FT3,FT4,TSH) and antithyroid autoantibodies (TgAb,TPOAb) were hospitalized into Affiliated Hospital,Jining Medical College between July 2009 and October 2013.The control group included 160 healthy subjects.We compared the prevalence of AITD between SLE patients and normal controls and also the positive rate of anti-thyroid autoantibodies was observed.We also compared the positive rate of anti-thyroid autoantibodies between AITD in SLE and simple SLE group and also analyzed the correlation between two groups of patients and SLE activity (evaluated by the titer of anti-dsDNA,C3,C4,CH50,SLEDAI score).Results Among them,45 patients suffered from AITD (20.5％).There were hyperthyroidism (n =6,13.3％) and hypothyroidism (including subclinical hypothyroidism) (n =26,57.8％),Hashimoto' s thyroiditis (n =13,28.9％).And 74 SLE cases were positive for anti-thyroid autoantibodies.The prevalence of AITD and the positive rate of anti-thyroid autoantibodies in SLE patients (20.5％,33.6％) were higher than that in normal controls (3.13％,7.50％)(P ＜ 0.05).The positive rate of anti-thyroid autoantibodies of SLE with AITD patients (62.2％) was higher than that in simple SLE (21.5％).No significant differences existed in anti-dsDNA titre,C3,C4,CH50 and SLEDAI score between two groups (P ＞ 0.05).Conclusions The SLE patients have a great prevalence of AITD and a positive rate of anti-thyroid autoantibodies.Those with anti-thyroid autoantibodies have a higher incidence of AITD and it has nothing to do with SLE activity.It is essential to monitor thyroid function and thyroid autoantibodies during the follow-ups.

Objective To investigate the time distribution characteristics and the epidemic trends of imported malaria cases in Yunnan Province. Methods The malaria case records and epidemiological history data of Yunnan Province were collected, and the local infection cases were excluded. The data were statistical analyzed. Results The imported malaria cases had a sig-nificantly seasonal periodicity(Q=26.574,P<0.05)and epidemic trends(Q=35.487,P<0.05). The imported peak was in May,while February was the lowest month of imported cases,and the difference was significant(Z=-2.619,P<0.05). The simple seasonal prediction model was the best model(R2= 0.677,BIC = 4.867)for forecast while the residual sequence was white noise(Q=14.226,P>0.05). By using the model to predict the cases in January,February and March of 2016,the num-ber(95％CI)were 29(7-50),22(0-44)and 31(8-54),and the actual number of imported malaria cases were 29,24 and 38 cases respectively and all cases were included in the 95％CI. Conclusion The imported malaria cases in Yunnan Province had a significantly seasonal periodicity and epidemic trends,and the established model has good prediction on the recent cases.

Objective To understand the status of Toxoplasma gondii infection in the population of Pu er City so as to pro-vide the evidence for formulating the strategy of toxoplasmosis control. Methods The population from Jingdong Jinggu and Menglian counties in Pu er City was surveyed IgG of T. gondii in serum was detected by ELISA. Results Totally 906 resident serum samples were detected and the IgG positive rate was 24.2%. The positive rates were higher in the aged groups of 30-39 years and 60-69 years and the difference among different aged groups was significant χ2=17.77 P 0.01 . There were no si-gnificant differences between different sexualities and among different educational levels and living habits P 0.05 . The posi-tive rates were 26.6% 194/730 15.5% 22/142 and 8.8% 3/34 in farmers students and other occupations respectively and there was a significant difference among them χ2=12.51 P 0.01 . The positive rates were 23.3% 198/849 and 36.8%21/57 in the farmers who had the habit of rearing pigs in pens and the farmers who had the habit of free ranging pigs respec-tively and there was a significant difference between them χ2=5.33 P 0.05 . Conclusion The IgG positive rate of T. go-ndii is very high in Pu er City and therefore the health education for toxoplasmosis control should be strengthened.

Objective To analyze the polymorphism of histidine rich protein 2(HRP II)gene in Plasmodium falciparum (Pfhrp2)from falciparum malaria patients in Yunnan Province,so as to lay the foundation for studying the defection of antigen genes of Plasmodium. Methods The filter paper blood samples and related information of falciparum malaria cases reported were obtained in Yunnan Province from August 2012 to September 2015. Under the guidance of the specific primers,the exon2 regions in Pfhrp2 gene in P. falciparum from DNA samples were amplified by PCR,and the PCR products were sequenced. The sequences of exon2 region in Pfhrp2 gene were blasted by comparing with the reference sequences AY816237,AY816240,and AY816301. Next,the polymorphism of the sequence in exon2 region of Pfhrp2 gene was analyzed by MEGA 5.04 software. The conserved sites and genetic distances between sequences were calculated by using the software as well,and the clustering tree was drawn according to the genetic distances between the amino acid sequences. Results A total of 218 bloods samples from the falciparum malaria cases in 15 prefectures of Yunnan Province were collected,and the sources of infection included Yun?nan,Africa and Myanmar. The PCR results showed that the exon2 regions in Pfhrp2 genes of 155 samples were positive by am?plification and their products were sequenced successfully. The sequence analysis showed that the length range of the amino acid residues of exon2 region in Pfhrp2 gene was from 115 aa to 298 aa,the average length was 239.7 aa. There was no statistically significance among the means of the amino acid residues of the isolates from Africa( 239.9 aa),Myanmar(239.5 aa)and Yun?nan(241.6 aa)(F=0.025,P>0.05). All the 155 amino acid sequences ended with type 12 repeat,98.1%(152/155)of them started with type 1 repeat and 1.9%(3/155)of them started with type 2. Type 2 presented most frequently repeat in all the se?quences and the average repeat times were 12.9. The homologous locus of the DNA sequences in exon2 regions of the 155 Pfhrp2 genes was 894 bp,among which the conservative sites accounted for 20.6%(186/894),and the variable sites for 78.2%(699/894). The genetic distances between the sequences of Africa isolates ranged from 0 to 0.741,and those of the Myanmar and Yun?nan isolates were 0-0.948 and 0-0.750,respectively. The cluster analysis showed that all the 155 sequences clustered into 3 cat?egories on genetic distances between amino acid sequences according to the size of the amino acid sequence length. At the same level,the sequences had approximate lengths and amino acid repeat types. Conclusion The sequence of exon2 region in Pfhrp2 gene of P. falciparum from falciparum malaria cases in Yunnan Province is highly polymorphic,the P. falciparum iso?lates are clustered mainly according to the size of the amino acid sequence of exon2 region in Pfhrp2 gene.

Objective To understand the geographical features of malaria in Yunnan Province,so as to provide the refer?ence for malaria elimination. Methods The data of malaria in Yunnan Province from 2012 to 2015 were collected and ana?lyzed. Results Totally 2 586 malaria cases were reported in Yunnan Province from 2012 to 2015,in which 274(10.60%) were local cases and 2 311(89.37%)were abroad imported,and one(0.03%)was domestic imported. The imported malaria cases and local cases were analyzed according to the sources and locations respectively,and the arithmetic means of the num?bers of imported and local cases were 96.29 and 10.96 respectively,the standard deviations of the numbers of imported and local cases were 421.18 and 19.12 respectively,and the difference of the means was not significant(Z=-0.326,P>0.10). Both the imported and local malaria cases could be clustered into five sections by the number of 5. The Herfendal?Hirshman indexes of the imported and local malaria cases were 8 121 and 1 598 respectively. Conclusions There is no significant difference of the distribution between the imported and local malaria cases,and they should be attaching equal importance. The non?uniform de?gree of imported cases is higher than that of the local cases,while both of them could be divided into five major clusters in the prevention and control work.

Objective:To investigate serum C-reactive protein (CRP) , procalcitonin (PCT) and neutrophil CD64 in predicting early infection after internal fixation of limb fractures.Methods:A total of 2 572 patients with limb fractures undergoing internal fixation in Taishun County People’s Hospital from January 2016 to December 2022 were enrolled. Postoperative infection occurred in 121 cases (infected group) and did not occur in 2 451 cases (uninfected group). Serum levels of PCT, CRP and CD64 were tested at admission and d1, d3, d5 and d7 after operation. Repeated measurement analysis of variance was used to compare the serum levels of PCT, CRP and CD64 at different time points between two groups, the receiver operating characteristic curve (ROC) was used to evaluate the predictive value of each index or in combination for early infection after internal fixation of limbs fractures.Results:The early infection rate after internal fixation was 4.70% (121/2 572). The levels of PCT, CRP and CD64 in the infection group began to rise after operation and decreased on d7; there were significant differences in PCT, CRP and CD64 levels between the two groups on d3, d5 and d7 after operation (PCT: Ftime=678.607, Fintergroup=2 218.323, Finteraction=653.150; CRP: Ftime=392.724, Fintergroup=1 812.502, FInteraction =379.577; CD64: Ftime=373.686, Fintergroup=4 817.438, Finteraction=528.353, all P< 0.001) . The area under the ROC curve of combined detection of PCT, CPR, and CD64 for predicting early infection was 0.856; the sensitivity and specificity of combined detection were 69.2% and 94.7%, respectively. The combined detection of three indicators showed better prediction values than PCT, CRP and CD64 alone ( Z=6.176, 3.838 and 2.431, P<0.01 or <0.05), and also better than combined detection of PCT and CRP ( Z=2.875, P=0.019). Conclusions:The combined detection of CD64, PCT and CRP is of value in prediction of postoperative infection after internal fixation of limb fractures, which is worthy of clinical application.

A series of farnesylthiosalicylamide derivatives(9a-9k)were designed and synthesized via condensating the carboxyl of farnesylthiosalicylic acid(FTS)with different amines, and the in vitro biological activity was evaluated. It was observed that eight of them displayed strong antitumor activity against five human cancer cells in vitro. Notably, compound 9k exhibited the most active with the IC50 values of 6. 05-8. 16 μmol/L range against the tested cancer cells, which were superior to those of FTS and even comparable to that of sorafenib in vitro. In addition, compound 9k down-regulated the protein of Bcl-2 and increased the expression levels of Bax and caspase-3 proteins, indicating that compound 9k could induce tumor cell apoptosis.

Objective:To investigate the application value of dual-energy CT in the differential diagnosis of lung metastases and benign nodules in breast cancer.Methods:The data of 96 patients with pathology-confirmed breast cancer at the Fifth Affiliated Hospital of Wenzhou Medical University from March 2017 to June 2021 were analyzed retrospectively. All patients received dual-energy chest CT scans within 2 weeks before surgery. All 96 patients were female, aged 31-84 (56±12) years. A total of 207 pulmonary nodules from 96 patients were classified into 81 lung metastases and 126 benign nodules according to pathological findings. Conventional CT features [longest diameter, boundary, location and CT value difference between arterial and venous phases (ΔCT) of nodules] and dual-energy CT parameters [standardized iodine concentration (NIC), slope of energy spectrum (λ HU) and normalized effective atomic number (nZ eff) in arterial and venous phases] were analyzed and measured. The χ 2 test, independent samples t test and Kruskal-Wallis rank-sum test were used to analyze the differences of conventional CT features and dual-energy CT parameters between lung metastases and benign nodules. First, the least shrinkage and selection operator (LASSO) regression method was used to screen conventional CT features and dual-energy CT parameters, and then logistic regression analysis was performed to screen out independent risk factors for lung metastases. Receiver operating characteristic (ROC) curves were used to evaluate the efficacy of CT parameters alone and logistic model in differentiating lung metastases from benign lung nodules. Results:There were statistically significant differences between lung metastases and benign nodules in longest diameter, ?CT, NIC, λ HU and nZ eff in arterial and venous phases (all P<0.05). LASSO regression and binary logistic regression analysis showed that the venous phase λ HU (OR=59.413, 95%CI 14.233-248.002, P<0.001) and the venous phase nZ eff (OR=4.508, 95%CI 2.787-7.290, P<0.001) were independent risk factors for predicting lung metastases. Among them, the venous phase λ HU had the highest diagnostic efficiency, with an area under curve (AUC) of 0.794 and an accuracy of 74.88%. The AUC of the logistic model constructed by combining the venous phase λ HU and the venous phase nZ eff could reach 0.958, and the accuracy was improved to 92.27%, which was significantly higher than the efficacy of the two alone ( Z=6.02, 9.54, all P<0.001). Conclusion:Dual-energy CT has great application value in the identification of lung metastases and benign nodules in patients with breast cancer, especially when combined with venous phase λ HU and venous phase nZ eff, the diagnostic efficiency is further improved.