[Analysis on mortality and premature death rates of 4 major chronic diseases in Ji'nan, 2015-2020].

Objective: To understand the characteristics and trend of the premature death rate of 4 major chronic diseases in Ji'nan from 2015 to 2020. Methods: The death cause surveillance data and population data during 2015-2020 in Ji'nan were collected, and abbreviated life table, Joinpoint regression analysis and other methods were used to analyze the characteristics and change trends of the premature death rates of 4 major chronic diseases. Results: The crude mortality rate and age standardized mortality rate changes for the 4 major chronic diseases from 2015 to 2020 range from 568.65/100 000 to 604.06/100 000 and 366.77/100 000 to 432.48/100 000, respectively. The annual premature death rate of 4 major chronic diseases declined by 3.33% averagely from 2015 to 2020 (95%CI: -6.25%--0.32%), which might be explained by the declines of the premature death rates of cardiovascular and cerebrovascular diseases [average annual percentage change (AAPC)=-3.23%, 95%CI: -6.32%--0.05%] and cancer (AAPC=-3.58%,95%CI:-6.83%--0.21%). The average decline rate in women (AAPC=-4.19%,95%CI:-7.56%- -0.70%) was higher than that in men (AAPC=-2.92%,95%CI: -5.65%--0.11%). Conclusions: The premature death rate of 4 major chronic diseases showed a downward trend in Ji'nan from 2015 to 2020. Men should be considered as a key population in the prevention and control of 4 major chronic diseases, and attention should also be paid to the non-significant declines in the premature death rates of chronic respiratory diseases and diabetes.

Primary intrasellar schwannoma with intratumoral hemorrhage mimicking pituitary apoplexy: A case report.

Tumours growing in the sella turcica are mostly pituitary adenomas. We describe a rare case of primary intrasellar schwannoma with intratumoral bleeding. A 38-year-old man presented with headache in association with bilateral supratemporal quadrantopsia. MRI showed an intrasellar mixed signal mass lesion with suprasellar extension. The majority of the pituitary hormones were normal. He was diagnosed as non-functional pituitary adenoma with pituitary apoplexy. Subtotal resection was achieved eventually via an endoscopic transnasal trans-sphenoidal approach. The histopathologic diagnosis was schwannoma. It is the first intrasellar schwannoma with intratumoral hemorrhage in literature to date. It implied that the primary intrasellar schwannoma has potential risk of intratumoral bleeding, which should be considered in the differential diagnoses of sellar lesions.

Effective Condition for Whole Testis Cryopreservation of Endangered Miho Spine Loach (Cobitis choii) Through the Optimization of Mud Loach (Misgurnus mizolepis) Whole Testis Cryopreservation Condition.

　　BACKGROUND: Miho spine loach (Cobitis choii) is an endangered Korean endemic fish. Whole testis cryopreservation is a good way for species preservation, but needs to the sacrifice of a large number of fish to optimize the freezing condition. Considering this limitation, a surrogate fish species was used for the protocol development. OBJECTIVE: This study was to establish the effective condition for Miho spine loach whole testis cryopreservation by optimizing the conditions for whole testis cryopreservation in an allied species, mud loach (Misgurnus mizolepis). MATERIALS AND METHODS: The condition for whole testis cryopreservation was optimized in mud loach first, and then the optimal condition was applied to Miho spine loach testes. RESULTS: The optimal condition for mud loach testis cryopreservation consists of the freezing medium containing 1.3 M dimethyl sulfoxide, 6% fetal bovine serum and 0.3 M trehalose, -1 C/min cooling rate and 26 degree C thawing temperature, which also permits effective cryopreservation of Miho spine loach testes. CONCLUSION: An effective cryopreservation condition for whole testis of the endangered Miho spine loach has been established by using mud loach as a surrogate fish.

Derivation of the clonal-cell lines from Siberian sturgeon (Acipenser baerii) head-kidney cell lines and its applicability to foreign gene expression and virus culture.

This study was conducted to establish and characterize the clonal-cell lines from Siberian sturgeon Acipenser baerii head-kidney tissues and to evaluate its applicability as a research tool. From the culture of A. baerii head-kidney derived cells, 10 cell lines were established first and then eight clonal-cell lines were derived from clonal growth and colony expansion of two cell lines that showed significant high colony-forming ability. All eight clonal-cell lines were morphologically similar and grew stably under monolayer culture but their growth rates were significantly different. They possessed diploid DNA contents, expressed epithelial cell-related genes and showed strong anchorage dependency to substrates. When a clonal-cell line was transfected separately with three plasmid vectors including fluorescent reporter genes driven by cytomegalovirus, marine medaka Oryzias dancena ß-actin or A. baerii ß-actin promoter, the cell lines expressed fluorescent signals regardless of promoter types. The cells harbouring foreign genes could be expanded to stable cell lines under drug selection and then they additionally could form the extensively proliferating colonies at low-density culture. Finally, the clonal-cell lines showed the susceptibility to viral haemorrhagic septicaemia virus (VHSV). Collectively, the clonal-cell lines from A. baerii head kidney were established and these cell lines will be able to provide an excellent in vitro system for various biological studies in this fish species.

Spirometra erinaceieuropaei severely infect frogs and snakes from food markets in Guangdong, China: implications a highly risk for zoonotic sparganosis.

Sparganosis is a parasitic disease caused by plerocercoid larvae of the genus Spirometra. In China, the main source of sparganosis is from Guangdong, 16.1% of the country's human sparganosis cases occur in this province. Frequent international trade of amphibians and reptiles in Guangdong may introduce new species of Spirometra into the local market. In this study, a large-scale, high-intensity sampling survey was conducted to find out the causative species and epidemic situation of Sparganosis in Guangdong. The prevalence of sparganum infection in five species of frogs (Boulengerana guentheri, Fejervarya multistriata, Hoplobatrachus chinensis, Pelophylax nigromaculatus and Quasipaa spinosa) and nine species of snakes (Elaphe carinata, Lycodon rufozonatum, Hypsiscopus plumbea, Ptyas dhumnades, P. korros, P. mucosa, Naja atra, Sinonatrix annularis and Xenochrophis piscator) was investigated in Guangdong, Southern China from May 2014 to August 2015. The results showed that 9.8% (50/511) of the frogs and 40.8% (141/ 346) of snakes were found to be infected by plerocercoids (spargana). To identify the species of the collected spargana, a partial sequence of the mitochondrial cytochrome c oxidase subunit1 gene (cox1) was amplified and sequenced. Phylogenetic analysis identified all the spargana specimens as Spirometra erinaceieuropaei. Our study indicated that S. erinaceieuropaei, a highly pathogenic parasite, is the only causative agent of sparganosis in Guangdong, China. This study suggests that the large numbers of frogs and snakes in food markets in Guangdong may impact public health in China by transmitting S. erinaceieuropaei sparganum. Additional steps should be considered by the governments and public health agencies to prevent the risk of food-associated Spirometra infections in humans in China.

Coexistence of intracranial aneurysm and hemangioblastoma: A case report and literature review.

The association of intracranial aneurysm and hemangioblastoma is extremely rare. This report regards a patient affected by Von Hippel-Lindau syndrome with multiple hemangioblastoma and two intracranial aneurysms, of which one was on a hemangioblastoma feeder vessel and the other on an unrelated vessel. Review of the literature revealed 13 other previously reported cases. Possible mechanisms to explain the association are discussed.

[Impact of heat waves on non-accidental deaths in Jinan, China].

OBJECTIVE: To assess the impact of heat waves on non-accidental deaths, and to investigate the influencing factors for deaths caused by heat waves in Jinan, China. METHODS: Daily death data and meteorological data for summer days with or without heat waves in Jinan from 2012 to 2014 were collected, and a cross-over analysis was conducted to evaluate the influence of heat waves on non-accidental deaths and deaths caused by other reasons. The univariate and multivariate logistic regression models were used to investigate the influencing factors for deaths caused by heat waves. RESULTS: The risks of non-accidental deaths and deaths caused by circulation system diseases during the days with heat waves were 1.82 times(95% CI: 1.47~2.36) and 1.53 times(95% CI: 1.14~2.07) those during the days without heat waves. The multivariate logistic regression analysis showed that old age(≥75 years)(OR=1.184, 95% CI: 1.068~1.313), low educational level(OR=1.187, 95% CI: 1.064~1.324), and deaths outside hospital(OR=1.105, 95% CI: 1.009~1.210) were associated with the high risk of deaths during the days with heat waves. CONCLUSION: Heat waves significantly increase the risk of non-accidental deaths and deaths caused by circulation system diseases in Jinan, and the deaths during the days with heat waves are related to age, educational level, and place of death.

[Under hypoxia condition contactin-1 regulates migration of MKN45 cells through RhoA pathway].

Recent studies have suggested that contactin-1 has a key role in cancer cell proliferation and migration, however the detailed mechanism of this process is still unclear. Here, human gastric cancer cell line MKN45 was employed. It was found that under hypoxia conditions contactin-1 mRNA and protein levels were both up-regulated by HIF-1alpha expression. Furthermore, although hypoxia increased the migration rate of MKN45 cells, contactin-1 (CNTN1) shRNA reversed this process. Meanwhile, RhoA V14 and RhoA V14N19 mutation constructs were employed, and it was found that constitutively active form of RhoA reversed the cell migration suppression induced by contactin-1 knockdown, while dominant-negative form of RhoA blocked hypoxia induced hypermigration. Apart from this, contactin-1 displayed the ability to phosphorylate the RhoA activator p115 RhoGEF. Thus, under hypoxia conditions, elevated HIF-1alpha seems to up-regulate contactin-1 expression and by this activate RhoA and facilitate migration of cancer cells.

Determination of feeder cell-based cellular niches supporting the colonization and maintenance of spermatogonial stem cells from prepubertal domestic cat testes.

Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long-term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC-specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome-shaped morphology, AP activity and expression of SSC-specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.

Establishment of a basic method for manipulating preantral follicles: effects of retrieval method on in vitro growth of preantral follicles and intrafollicular oocytes.

The aim of this study was to establish a basic manipulation protocol of preantral follicles for deriving developmentally competent oocytes. Primary, early and late secondary follicles retrieved from the ovaries of 14-day-old F1 (C57BL/6 x DBA2) female mice mechanically or enzymatically were cultured singly and in vitro growth of the follicles and maturation of intrafollicular oocytes were subsequently monitored. A mechanical method retrieved more (p < 0.0001) follicles (339 +/- 48 vs. 202 +/- 28) than an enzymatic method. However, the enzymatic method collected more singly isolated follicles that could be provided for subsequent culture (102 +/- 26 vs. 202 +/- 28). When an enzymatic method was employed, early and late secondary follicles required 9 and 6 days for reaching the maximal incidence of the pseudoantral stage. However, primary follicles were not possible to develop into the pseudoantral stage. The optimal duration of oocyte maturation from the onset of follicle culture was 7 days and 5-7 days for early and late secondary follicles, respectively. A general decrease in oocyte diameter (65.2-65.53 microm vs. 75 microm) and zona thickness (5.41-5.74 microm vs. 7.76 microm) was detected in in vitro-derived compared with in vivo-derived matured oocytes. Pronuclear formation was detected in 86-94% of mature oocytes after parthenogenetic activation and no significant difference was detected among groups. These results showed that preantral follicles retrieved by an enzymatic method underwent step-by-step growth in vitro, which could yield mature oocytes.