BACKGROUND: Transplantation of fragments of ovarian cortex is performed without vascular reanastomosis, therefore, to increase the tolerance of ovarian tissue to the freezing-thawing and ischemic injuries is critical for follicular survival and functional longevity of the graft.OBJECTIVE: To investigate the effect of follicle stimulating hormone (FSH) on the morphological and function of sheep ovary tissue in the process of cryopreservation, so that to provide new freezing method for human ovary tissues. METHODS: Healthy BALB/c strain female nude mice were equally and randomly divided into 3 groups and subjected to heterotopic transplantation of fragments of sheep ovarian cortex. (1) Control group: transplantation following sampling; (2) experimental group: transplantation following freezing-thawing, and the solution was free of FSH; (3) FSH group: the freezing and thawing and culture fluid contained FSH. The estrous cycle recovering rate, estrous cycle recovering time, the number of follicle were observed following transplantation. The histological changes, and the level of E2 in blood serum were observed at 4 weeks after transplantation.RESULTS AND CONCLUSION: There were no significant differences in estrous cycle recovering rate and number of follicle/PHF between FSH and control groups (P> 0.05), but the number of follicle was greater than the experimental group (P< 0.05). Moreover, the time of estrous cycle recovering in FSH group was similar to control group, but shorter than experimental group (P < 0.05). There were more developing follicular in FSH group, but few in experimental group at 4 weeks. No significant difference was detected in E2 level in blood serum between FSH and control group (P > 0.05), but significantly greater than the experimental group (P< 0.05). Results show that FSH addition in vitrification fluid can improve ovarian follicle survival following cryopreserved sheep ovary tissue transplantation.

The emergence of atypical El Tor strains from V .cholerae in South Asia and Africa has been attributed to several outbreaks in recent decades ,however ,backgrounds of such strains in China remain exclusive .In this study ,PCR am-plification of both El Tor and classical alleles for ctxB ,tcpA ,rstR and hlyA genes was attempted in sixty-nine El Tor isolates from Fujian between 1962 to 2005 ,in addition ,some amplicons were sequence-analyzed .Thus ,the time point of atypical EVC strains in Fujian was determined ,genetic diversities of such strains were investigated .It was revealed that ctxB-Cl ,tcpA-Cl and hlyA genes were detected in O1 serogroup EVC isolates from Fujian since 1962 .Although rstR-Cl gene was solely detected in isolates between 1994 to 2000 .It was indicated by sequence analysis that atypical EVC strains from Fujian possessed a novel T→G mutation at residual 204 of the ctxB gene .Remarkably ,two novel ctxB genotypes (ctxB-10 and ctxB-11) were identified in one strain .The residual 115-C of ctxB in ctxB-11 showed characteristics of ctxB-Cl ,however ,its residual 203-T demonstra-ted characteristics of ctxB-El .This observation implied that it was common in O1 serogroup EVC strains from Fujian hybrid-ized with classical alleles since 1962 ,which would be the earliest time-point for the emergence of atypical El Tor strains hitherto in literature .Emergence of atypical El Tor strains harboring rstR-Cl in Fujian occurred since 1994 .Meanwhile ,novel mutation sites and ctxB genotypes were observed in Fujian isolates ,including diverse combination of ctxB genotypes in one strain and combination of biotype-specific sites in ctxB sequences .In summary ,molecular characterization of O 1 serogroup EVC strains from Fujian was unique and geography-associated .

Objective To study the effect of Chinese medicinal compound recipe ( modified Fuzheng Yiliu Decoction, MFYD) on the proliferation of colon cancer HT-29 cells and on the expression of cellular apoptosis gene, Bcl-2, and tumor-inhibiting gene, P53. Methods Colon cancer HT-29 cells were divided into serum containing MFYD groups ( treated with different concentrations of serum containing MFYD) and blank serum group. Methyl thiazolyl tetrazolium ( MTT) , real time cellular analysis ( RTCA) and reverse transcription poly merase chain reaction ( RT-PCR) were used to detect the cell proliferation and mRNA expression of Bcl-2 and P53 separately. Results The proliferation of colon cancer HT-29 cell line was inhibited by MFYD ( P<0.05) in concentration-and time-dependent manner. The Bcl-2 mRNA expression was decreased and p53 mRNA was increased markedly in HT-29 cells after co-culturing with 12% or 15% volume fraction of serum containing MFYD for 48 hours, the differences being significant compared with the blank serum group ( P<0.05). Conclusion MFYD has obvious inhibitory effect on the proliferation of colon cancer HT-29 cells, and its mechanism may be related to down-regulating the expression of Bcl-2 gene and up-regulating the expression of P53 gene.

Objective To evaluate the feasibility of individual template assisting permanent interstitial brachytherapy for skull base region.Methods From August 2010 to June 2012,20 patients with recurrent malignancies of skull base region received 125I seed permanent implantation.Before implanting,all patients underwent CT scan,the data were stored in Dicom format and imported into brachytherapy treatment planning system (BTPS).The preplan were finished by BTPS.Then,the digital model based on virtual needle and skin CT slices was reconstructed by Mimics and Geomagic.According to the digital model,the individual template was made from medical light-cured resin using rapid forming machine.During implanting,needles were inserted under the individual template assistance and 125I seeds were implanted according to preplan.After implanting,implantation quality was evaluated by 2D and 3D CT images and BTPS.The needles and seed distribution were observed.The actual dose distribution of target volumes D90,V100 and V150 was calculated.The complications were recorded within 72 hours.Results Interstitial implantation of 20 patients were performed successfully and efficiently under the individual template assistance without serious complications.Because of containing the virtual needle positioning and face surface feature information at the same time,the individual template significantly improved precision of needle location and orientation.The actuarial median number of 125I seeds implanted was 70 (range,20-172),and actuarial median D90,V100 and V150 was 181.6 Gy (127.4-279.6 Gy),98.2％ (94.6％-100％),43.2％ (24.3％-52.2％),respectively.The seed distribution and dosimetric quality were well controlled.Conclusions The method of individual template assisting permanent interstitial brachytherapy for skull base region is feasible,which can improve the accuracy of needle position placement.

In order to investigate EV71 antibody levels among general population from Fujian Province after the 2008‐2009 HFMD epidemics ,390 sera‐specimens were collected from 390 participants in 2010 .EV71‐specific antibody was detected by neutralization test ,indicating 186 (47 .69% ) sera of 390 were EV71‐seropositive .Although the difference by gender was not statistically significant on positive rates and antibody titers ,significant differences were observed in positive rates and antibody titers among age groups .The positive rate was increasing with age ,while the 0 age‐group yielded the lowest positive rate of 16 .67% .Subsequently ,significant difference was detected among positive rates and antibody titers between age groups of 0 to 4 years‐old and 5‐years‐old ,with the positive rate of 25 .33% and 61 .67% ,respectively .Therefore ,the EV71 antibody levels among general population from Fujian Province after the 2008‐2009 HFMD epidemics was still in the low level ,especially the age groups of 0 to 4 years‐old .The epidemic of HFMD mainly caused by EV71 will still occur in the future ,and children under 5 years old are major susceptible population ,continuously intensive surveillance ,prevention and control are required .

Objective To establish a Chinese hamster model of babesia infection, to find the changing pattern of organs and biochemical parameters in Chinese hamster infected with Babesia, and to promote the detection and treatment of babesiosis.Methods Healthy 5-week old Chinese hamsters were infected by intraperitoneal injection of blood containing Babesia.Blood samples were collected at 0, 2, 4, 6, 8, 10, 12, 14, 16, 23, 30, and 37 days after infection from 5 hamsters at each time point.Blood smears were prepared to detect the parasites using Giemsa staining.ELISA assay was employed to test the IL-2 concentration.The blood biochemical indexes were detected using an automatic biochemical analyzer.DNA was extracted from the whole blood and REAL-TIME RCR was performed to determine the reproduction of Babesia.Aftert the animals were sacrificed, the heart, lung, spleen, liver, and kidney were taken to analyze the changes of organ coefficients.Results The highest level of Babesia in the hamsters occurred on day 4 after the Babesia injection, and then showing a decreasing tendency.However, there was a transient increase on the 12th day after infection.The liver and spleen displayed most extensive response to the infection showing hepatomegaly and splenomegaly, but the variation of heart and kidneys coefficients was within the norm.There were prominent changes of blood cells, especially leucocytes, with two peaks at day 10 and 23 after the Babesia infection.The peak changes of blood biochemical indexes occurred at day 12 after infection.The concentration of serum IL-2 reached a peak on the 10th day after infection.Conclusions The Chinese hamsters display typical characteristics of tick-borne diseases such as hepatomegaly and splenomegaly.The immunological system is activated along with the infection and reaches a highest stage in the second week.Afterwards the Babesia can live in the hamster body for a long period of time.The results of this study provide useful information supporting further studies on the detection, treatment and prevention of Babesiosis.

ObjectiveTo investigate emergency diagnosis and treatment of pelvic fracture complicated with traumatic rupture of urethra and bladder,and to improve the success rate of treatment on pelvic fracture.MethodsClinical data of 52 cases of pelvic fracture complicated with traumatic rupture of urethra and bladder in department of emergency and urology from 2000 to 2010 was retrospectively analyzed.Results Among the 52 patients,there was 41 cases of pelvic fracture complicated with posterior urethral disruption,15 cases complicated with rupture of bladder and 4 cases complicated withtraumatic rupture of urethra and bladder at the same time.In 41 cases with posterior urethral rupture,6 individual's condition were relatively so severe that they onlyunderwent bladder puncture nephrostomy,and 29 cases underwent traction urethral realignment,the other 6 cases didn't undergo surgery; In 15 cases of patients with bladder rupture,2 patients were performed urethral realignment and bladder repair,11 patients underwent the bladder repair only and the other 2 patients were not performed surgery.There were 8 patients died and the mortality rate was 15.4％.Six died cases failed to conduct emergency surgery because of uncontrollable bleeding and another 2 cases died due to multiple organ failure.ConclusionPelvic fractures is a disease with more complications,it should be diagnosed as early as possible.Patients invalid for conventional anti-shock should be performed pelvic external fixation and emergency embolization to stop bleeding in the emergency department,and undergo associated processing after they are in stable condition.

Objective:To investigate the relationship between cognitive function and brain event-related potential in patients with lacunar cerebral infarction.Methods:A total of 464 patients with lacunar cerebral infarction admitted to the Department of Neurology, Kailuan General Hospital from 2014 to 2019 were prospectively selected as observation subjects (case group). According to mini-mental state examination (MMSE) score, the patients in the case group were divided into 352 cases of lacunar cerebral infarction with normal cognition and 112 cases of mild cognitive impairment. At the same time, 100 healthy volunteers were selected as the control group. All subjects were assessed by simple intelligent mental state, Zung self-rating anxiety scale, Zung self-rating depression scale and brain event-related potential P3a and P3b. The measurement data of normal distribution adopts one-way ANOVA, the measurement data of non normal distribution adopts Kruskal Wallis H test, and the counting data adopts χ2. Multivariate statistical analysis was performed by unconditional Logistics (stepwise method). Results:The proportions of smokers in control group, lacunar cerebral infarction cognitive normal group and lacunar cerebral infarction mild cognitive impairment group were 20.00% (20/100), 38.07% (134/352) and 46.42% (52/112), respectively. The proportions of drinkers were 18.00% (18/100), 33.24% (117/352), 33.93% (38/112), respectively. The proportions of hypertension were 38.00% (38/100), 58.24% (205/352), 59.82% (67/112), respectively. The proportions of hyperhomocysteinemia were 19.00% (19/100), 34.00% (120/352) and 68.75% (77/112), respectively, and the differences among the three groups were statistically significant ( χ2 values were 15.66, 7.91, 11.86 and 54.57, respectively; P<0.001, 0.019, 0.003, <0.001). The peak latency CZ leads of visual P3b wave group N2 were (271.48±40.65), (285.67±44.08) and (290.57±68.41) ms, respectively. PZ leads were (276.70±50.92), (287.86±43.28) and (312.16±62.75) ms. P3b peak latency FZ leads were (392.67±42.50), (405.82±52.43) and (410.34±64.27) ms. CZ leads were (395.04±42.44), (412.51±55.86) and (433.28±66.32) ms. PZ leads were (398.24±40.93), (411.17±49.48) and (435.78±67.69) ms. N2 amplitude CZ leads were (-3.99±2.81), (-3.60±3.00) and (-2.70±2.37) μV, PZ leads were (-3.18±2.69), (-2.91±2.62) and (-1.87±2.89) μV, respectively. Leads P3b amplitude of FZ were 5.27 (3.27, 7.40), 4.21 (2.31, 6.49) and 3.12 (1.61, 5.08) μV. CZ leads were 4.81 (2.78, 6.71), 4.15 (2.76, 6.16) and 3.51 (1.75, 5.15) μV. PZ leads were 5.17 (3.03, 6.97), 4.40 (2.89, 6.12) and 3.43 (1.52, 5.34) μV. There were statistically significant differences among the 3 groups ( F=3.29, 14.49, 3.95, 11.73, 14.06, 5.66 and 3.57, H=18.23, 10.33,18.25; P=0.027, <0.001, 0.025, <0.001, <0.001, 0.004, 0.042, <0.001, 0.006, <0.001). The peak latency FZ leads of visual P3a wave group N2 were 265.00 (256.00, 286.00), 277.00(260.00,300.00), 282.00(270.00,304.00) ms, respectively. CZ leads weres 274.00(255.00,305.00), 285.00(262.00,329.00), 293.50(270.00,346.00) ms. P3a peak latency FZ leads were (413.83±49.58), (429.83±55.38) and (449.04±54.79) ms, CZ leads were (441.53±61.78), (457.12±69.29) and (460.23±72.24) ms. PZ leads were (430.14±54.53), (462.31±69.2) and (470.02±74.92) ms. N2 amplitude FZ leads were (-6.34±3.13), (-5.72±2.96) and (-4.92±2.05) μV, respectively. Leads P3a amplitude of FZ were 4.00 (2.28, 5.55), 3.15 (2.14, 4.91) and 2.80 (2.19, 4.19) μV. CZ lead were 3.37 (1.98, 4.66), 2.73 (1.70, 3.97) and 2.41 (1.64, 3.45) μV. There were statistically significant differences among the three groups ( H=13.92, 8.65, 9.17, 10.02, F=8.18, 6.33, 10.73, 4.62, P =0.001,0.013,0.010,0.007, <0.001,0.002, <0.001,0.010). Logistic regression analysis showed that alcohol consumption, P3b peak latency and wave amplitude PZ lead, N1 wave amplitude of visual P3a group FZ lead were the influencing factors of MMSE ( OR=0.04, 1.01, 0.76, 1.51, 95% Cl were 0.00-0.30, 1.00-1.03, 0.59-0.97, 1.08-2.10, P=0.002,0.007,0.029,0.016). Conclusion:The peak latency and amplitude of endogenous psychological cognitive potentials N2, P3b and P3a of event-related potentials P3b and P3a in patients with lacunar cerebral infarction were prolonged and decreased. At the same time, with the occurrence of clinical cognitive impairment, the peak latency and amplitude of these cognitive potentials were further prolonged and decreased more significantly. Alcohol consumption, P3b peak latency and PZ lead of visual P3b wave group, and FZ lead of N1 wave of visual P3a wave group were the influencing factors of simple intelligent mental state.

Objective:To screen and analyze ferroptosis-related genes (FRG) impacting the prognosis of colorectal adenocarcinoma patients based on bioinformatics.Methods:RNA sequencing data including the clinical information of 545 colorectal adenocarcinoma patients and 602 data sets were downloaded from the Cancer Genome Atlas (TCGA) database. FRG gene sets were downloaded from FerrDb database. FRG expression dataset could be obtained after taking the intersection between FRG gene sets and TCGA database gene sets. Differentially expressed FRG and prognosis-related genes between colorectal adenocarcinoma tissues and the adjacent tissues were screened by using R software, and finally FRG influencing the prognosis of colorectal adenocarcinoma were obtained. According to protein-protein interaction networks, the interaction and the expression association of proteins were analyzed. LASSO regression analysis was used to build a risk model for patients' 5-year overall survival rate. The risk value was calculated for 509 colorectal adenocarcinoma samples in the TCGA database, and then the median risk value was taken as the cut-off value. All patients were divided into the high-risk group (≥ median risk value) and the low-risk group (< median risk value), and the survival curves of the two groups were drawn. The receiver operating characteristic (ROC) curve was drawn for predicting the 5-year overall survival rate of colorectal adenocarcinoma patients in a time-dependent way in TCGA database according to the risk value of FRG prognosis model. Cox proportional risk model was used to make univariate and multivariate survival analysis in order to screen factors affecting the prognosis. The pathway enrichment analysis of prognosis-related FRG of colorectal adenocarcinoma was performed based on gene ontology (GO) database and Kyoto Encyclopedia of Genes and Genomes (KEGG) database.Results:The clinical information of 545 patients and 602 datasets were extracted from the database. A total of differential expressed 199 FRG in colorectal adenocarcinoma and 28 prognosis-related FRG were identified. After taking the intersection, 21 FRG affecting the prognosis of colorectal adenocarcinoma patients were identified. DUOX2, NOX4, NOX1, DDIT3, JDP2, ATP6V1G2, ULK1, ATG3 were probably associated with WIPI1; expressions of NOX4, NOX5, PLIN4 were positively correlated with ATP6V1G2, while the expression of ULK1 was negatively correlated with MAPK1, MYB, FANCD2, ATG3 and ATP5MC3. LASSO regression analysis showed that 15 FRG were finally screened out (ATP5MC3, NOX4, NOX5, ALOX12B, ATG3, WIPI1, MAPK1, MYB, AKR1C1, DDIT3, JDP2, ATP6V1G2, DRD4, SLC2A3, PLIN4), and the risk model was constructed by calculating the risk value, and the risk value = NOX4×0.139-ATP5M3×0.108+NOX5×1.486+ALOX12B×0.475-ATG3×0.030-WIPI1×0.170-MAPK1×0.271-MYB×0.063+AKR1C1×0.021+DDIT3×0.186+JDP2×0.292+ATP6V1G2×0.777+DRD4×0.294+SLC2A3×0.059+PLIN4×0.113. The overall survival of patients in the high-risk group was worse than that in the low-risk group ( P < 0.001). The 5-year overall survival rate was 48.2% in the high-risk group and 76.8% in the low-risk group. Multivariate survival showed that the age and risk value were independent affecting factors of the prognosis. ROC curves revealed that the risk model constructed by using prognosis-related FRG could well predict the 5-year overall survival rate of patients (the area under the curve was 0.728). The differential expressed genes of both groups may be associated with genetic pathways such as extracellular matrix composition, extracellular structure composition and focal adhesion. Conclusions:The prognostic risk model constructed by the screened FRG can better evaluate the prognosis of colorectal adenocarcinoma patients. These FRG are expected to become new candidate biomarkers related to the prognosis of colorectal adenocarcinoma.

OBJECTIVEUsing heteroduplex mobility assay (HMA) to subtype human immunodeficiency virus type 1 (HIV-1) for the purpose of understanding HIV-1 subtype epidemic in Fujian province.

METHODSDNA fragments of HIV-1 env gene were amplified from peripheral blood mononuclear cell (PBMC) cocultures of HIV-1 infected individuals by nested polymerase chain reaction (PCR). Heteroduplexs were formed through hybridizing PCR products from the samples and reference plasmid. According to the mobility of heteroduplexs in polyacrylamide gel electrophoresis, HIV-1 subtype from that sample was characterized and further confirmed by nucleotide sequencing analysis.

RESULTSThirteen of 15 (86.67%) samples were successfully subtyped by HMA, except 2 failures. Subtype E and B took up 80% (12/15) and 6.67% (1/15) respectively. Results indicated a high concordance between HMA and nucleotide sequencing analysis and concordance rate was 86.67% (13/15).

CONCLUSIONSSubtype E appeared to be the major epidemic strain of HIV-1 in Fujian. HMA showed the characteristics of fastness, easiness, economic and with high specificity, and can be used in the surviellance for the epidemic strain of HIV-1.