As a typical human pathogenic fungus,

Pluripotent stem cells (PSCs) can be expanded in vitro in different culture conditions, resulting in a spectrum of cell states with distinct properties. Understanding how PSCs transition from one state to another, ultimately leading to lineage-specific differentiation, is important for devel-opmental biology and regenerative medicine. Although there is significant information regarding gene expression changes controlling these transitions, less is known about post-translational modifi-cations of proteins. Protein crotonylation is a newly discovered post-translational modification where lysine residues are modified with a crotonyl group. Here, we employed affinity purification of crotonylated peptides and liquid chromatography–tandem mass spectrometry (LC–MS/MS) to systematically profile protein crotonylation in mouse PSCs in different states includ-ing ground, metastable, and primed states, as well as metastable PSCs undergoing early pluripotency exit. We successfully identified 3628 high-confidence crotonylated sites in 1426 proteins. These crotonylated proteins are enriched for factors involved in functions/processes related to pluripotency such as RNA biogenesis, central carbon metabolism, and proteasome function. Moreover, we found that increasing the cellular levels of crotonyl-coenzyme A (crotonyl-CoA) through crotonic acid treatment promotes proteasome activity in metastable PSCs and delays their differentiation, consis-tent with previous observations showing that enhanced proteasome activity helps to sustain pluripo-tency. Our atlas of protein crotonylation will be valuable for further studies of pluripotency regulation and may also provide insights into the role of metabolism in other cell fate transitions.

As self-subunit swapping chaperones or metallochaperones, the activators assist nitrile hydratases to take up metal ions and they are essential for active expression of nitrile hydratases. Compared with nitrile hydratases, the activators have a low sequence identity. Study of the activation characteristics and the relationships between structures and functions of the activators is of great significance for understanding the maturation mechanism of nitrile hydratase. We co-expressed low-molecular-mass nitrile hydratase (L-NHase) from Rhodococcus rhodochrous J1 with four heterologous activators respectively and determined their activation abilities. Then we made sequence analysis and structure modelling, and studied the functions of the important domains of the activators. Results showed that all four heterologous activators could activate L-NHase, however, the specific activities of L-NHases were different after activation. L-NHase showed the highest specific activity after being activated by activator A, which was 97.79% of that of the original enzyme, but the specific activity of L-NHase after being activated by activator G was only 23.94% of that of the original enzyme. Activator E and activator G had conserved domains (TIGR03889), and deletion of their partial sequences resulted in a substantial loss of activation abilities for both activators. Replacing the N-terminal sequence of activator G with the N-terminal sequence of activator E, and adding the C-terminal sequence of activator E to the C-terminus of activator G could increase the specific activity of L-NHase by 178.40%. The activation by nitrile hydratase activators was universal and specific, and the conserved domains of activators were critical for activation, while the N-terminal domain and C-terminal domain also had important effects on activation.

Protein lysine methylation is a prevalent post-translational modification (PTM) and plays critical roles in all domains of life. However, its extent and function in photosynthetic organisms are still largely unknown. Cyanobacteria are a large group of prokaryotes that carry out oxygenic photosynthesis and are applied extensively in studies of photosynthetic mechanisms and environmental adaptation. Here we integrated propionylation of monomethylated proteins, enrichment of the modified peptides, and mass spectrometry (MS) analysis to identify monomethylated proteins in Synechocystis sp. PCC 6803 (Synechocystis). Overall, we identified 376 monomethylation sites in 270 proteins, with numerous monomethylated proteins participating in photosynthesis and carbon metabolism. We subsequently demonstrated that CpcM, a previously identified asparagine methyltransferase in Synechocystis, could catalyze lysine monomethylation of the potential aspartate aminotransferase Sll0480 both in vivo and in vitro and regulate the enzyme activity of Sll0480. The loss of CpcM led to decreases in the maximum quantum yield in primary photosystem II (PSII) and the efficiency of energy transfer during the photosynthetic reaction in Synechocystis. We report the first lysine monomethylome in a photosynthetic organism and present a critical database for functional analyses of monomethylation in cyanobacteria. The large number of monomethylated proteins and the identification of CpcM as the lysine methyltransferase in cyanobacteria suggest that reversible methylation may influence the metabolic process and photosynthesis in both cyanobacteria and plants.
Bacterial Proteins/metabolism* ; Lysine/metabolism* ; Methyltransferases/metabolism* ; Photosynthesis ; Protein Processing, Post-Translational ; Synechocystis/growth & development*

Objective: To analyze the epidemiological and spatial-temporal distribution of Brucellosis, epidemic encephalitis B and hemorrhagic fever with renal syndrome (HFRS) in Gansu province during 2014-2018 so as to provide evidence for the prevention and control of those diseases. Methods: A database was established in Gansu province from 2014 to 2018, using the geographical information system. A spatial distribution map was drawn, with trend analysis and space-time clustering used to study the 3-dimention of the diseases, by using both ArcGIS 10.5 and SaTScan 9.6 softwares. Results: Results from the trend surface analysis showed that the incidence of Brucellosis decreased gradually from north to south parts while the U type curve could reflect the distribution from the east to the west areas. Incidence of epidemic encephalitis B decreased significantly from south to north areas in the province, with incidence higher in the eastern than in the mid-west region. Difference on the incidence of HFRS was not significantly visible in the eastern and western regions, while the incidence was slightly higher in the southern than the northern parts of the province. Spatial and space-time clustering did exist among the 3 diseases in Gansu from 2014 to 2018. The areas with clusters of Brucellosis appeared in the eastern parts during 2014-2015, including 19 counties. The areas with secondary clusters of Brucellosis were seen in the Hexi district, including 4 counties, during 2017-2018. The areas with high incidence of epidemic encephalitis B were clustered in the middle and southeast areas, including 32 counties, during 2017-2018. Areas with most clusters of HFRS appeared in Min county of Dingxi city in 2018, with the areas of secondary clusters in 8 counties of the eastern areas in 2018. Conclusions The overall incidence rates of the 3 natural focus diseases were in a upward trend and showing obvious characteristics on spatial clustering. According to the distributive characteristics, effective measures should be developed accordingly.

Objective To explore the distribution of genetic structure of Y-SNP and Y-STR genetic markers in different ethnic groups and its application in forensic science. Methods SNaPshot minisequencing was used to detect the polymorphisms of 12 Y-SNP loci in 439 males from 6 ethnic groups, including Guangxi Han, Guangxi Jing, Guangxi Miao, Guangxi Yao, Guangxi Zhuang and Guangxi Dong. DNATyperTM Y26 kit was used to multiplex-amplify 26 Y-STR loci. The PCR products were analyzed by 3130xl genetic analyzer. The network analysis of Y-STR haplotype under the same Y-SNP haplogroup was analyzed by Network 5.0 software. Results Six haplogroups defined by 12 Y-SNP loci were detected in 6 ethnic groups, and 362 haplotypes were detected in 26 Y-STR loci. The haplotype diversity was 0.996 6. In the C haplogroup, the samples from Guangxi Yao, Guangxi Zhuang and Guangxi Dong were clustered on different branches; in the O1 haplogroup, those from Guangxi Zhuang, Guangxi Miao and Guangxi Jing were relatively independent and clustered separately; in the O2 haplogroup, some samples from Guangxi Miao and Guangxi Yao were gathered in a cluster. Conclusion Based on the Y-STR network analysis of samples with identical haplogroup of Y-SNP, some ethnic groups can be preliminarily distinguished, which could be used to infer male suspects' ethnic group through detecting their genetic markers left in the crime scene.
China ; Chromosomes, Human, Y ; Ethnicity ; Genetics, Population ; Haplotypes ; Humans ; Male ; Microsatellite Repeats

To establish the HPLC fingerprint and determine five index components (loganic acid， chlorogenic acid， loganin， sweroside and asperosaponin Ⅵ) of Zishen Yutai pills by high performance liquid chromatography, and provide a scientific basis for its quality control. The fingerprint chromatogram was analysed by the chromatographic fingerprint similarity evaluation system for tradition Chinese medicine (2012), fifteen common peaks were obtained at the wavelength of 254 nm. Different batches of Zishen Yutai pills showed a similarity of above 0.90 in HPLC fingerprint profiles. For the quantitive analysis method, The separation of five components showed good regression (>0.999 2) with linear ranges, and the mean recoveries were in the range of 97.62%-101.9%, with the RSD (=9) less than 3%. The established fingerprint and quantitative analysis methods are highly specific, simple and accurate, which can reflect the quality of Zishen Yutai pills more comprehensively, and can be used for its quality control.

Cell culture is an important tool for biological research.To better understand the pathogenesis and therapeutic methods of the tumors,the three-dimensional cell culture is applied by more and more researchers to create a culture environment that closes to the tumor microoenvironment.Thanks to the advances in the tissue engineering technology,many kinds of models of the three-dimensional cell culture achieve wide accessibility.Compared with the traditional two-dimensional cell culture,the three-dimensional cell culture is better in simulating physiological features of the human histology and cells,including cell proliferation and differentiation,the interaction of cell to cell and cell to matrix.This paper reviews the progress of multi-cellular tumor spheroids (MCTS) culture technique of the three-dimensional cell culture for treatment of bladder cancer.

Objective To understand the situations of departments of intensive care units (ICUs) of different level hospitals in Guizhou Province, and to provide directions and evidences for improving quality control in critical care medicine.Methods A county-level hospital and a provincial-level hospital's comprehensive ICU in Guizhou Province were selected to record and analyze and compare the structural indicators, patient admission and transfer, disease distribution, ventilator associated pneumonia (VAP), intravascular catheter related blood stream infection (CRBSI) and catheter-associated urinary tract infection (CAUTI) of the two hospitals' comprehensive ICU in 2017.Results The ICU of the People's Hospital of Suiyang County (county hospital) was found in 2012, and the ICU of the Affiliated Hospital of Guizhou Medical University (provincial hospital) was found in 1994. Until 2017, there were 10 and 46 beds, 6 (all of them hold bachelor's degree) and 18 physicians (6 of them hold PhD, 5 of them hold master's degree, 7 of them hold bachelor's degree), 17 (4 of them hold bachelor's degree, 13 of them hold college degree or graduated from secondary school) and 69 nurses (2 of them hold master's degree, 53 of them hold bachelor's degree, 14 of them hold college degree or graduated from secondary school) in the two ICUs respectively, there were significant differences in the education background of the physicians and nurses between the two ICUs (bothP < 0.01). During 2017, 471 cases were admitted to the ICU of the county hospital while 1633 cases were admitted to the ICU of the provincial hospital. Compared with the ICU of the provincial hospital, the ratio of the patients with acute physiology and chronic health evaluationⅡ(APACHE Ⅱ) ≥ 15 at admission was lower (74.8％ vs. 85.1％,P < 0.01), the ratio of direct admission was higher (30.8％ vs. 17.4％,P < 0.01), the ratio of the patients admitted to the ICU more than once was lower (0.8％ vs. 5.0％,P < 0.01), the ratio of the patients whose the length of ICU stay less than 24 hours was higher (51.6％ vs. 13.7％,P < 0.01), the ratio of the patients whose the length of ICU stay more than 28 days was lower (1.1％ vs. 2.9％, P < 0.05), the ratio of the patients discharged against-advice (25.5％ vs. 20.5％,P < 0.05) was higher, the ratio of the patients transferred to other hospitals was higher (5.1％ vs. 0.3％,P < 0.05), and the ICU mortality was lower (4.0％ vs. 13.9％,P < 0.01) in the ICU of the county hospital. The top three kinds of diseases treated in the ICU of the county hospital were brain injury (27.4％), trauma (19.1％) and toxication (6.8％); while in the ICU of the provincial hospital were brain injury (18.6％), sepsis (16.2％) and severe acute pancreatitis (4.8％). In addition, the incidences of VAP, CRBSI and CAUTI in the ICU of the county hospital were 10.0/1000 ventilator days, 1.4/1000 catheter days, 0.5/1000 catheter days; while in the ICU of the provincial hospital were 5.8/1000 ventilator days, 2.0/1000 catheter days, 3.7/1000 catheter days, respectively.Conclusions There are short of physicians and nurses in the ICU of the provincial and county hospitals in Guizhou Province, and the educational level of the medical staff in the ICU of the county hospital is relatively low. Moreover, there were significant differences in the admissions and treatments and the outcomes of the critically ill patients between the two ICUs. The characteristics of the ICUs of county hospitals should be fully considered when the quality control of critical care medicine and continuing medical education are done.

OBJECTIVETo investigate the clinical effect of Halo-vest head ring in the treatment of replantation of total scalp avulsion.

METHODSWe treated 11 cases of total scalp avulsion with the anastomosis of arteriovenous vessels and Halo-vest head ring from December 2006 to February 2015.

RESULTSOne patient's replanted scalp got necrosis because of serious contusion which was healed without hair growth after free skin graft and dressing. All the scalp flaps in the other 10 patients survived. After 3-96 months follow-up, the wound completely healed, the scalp and hair grew well with satisfactory appearance.

CONCLUSIONSThe use of Halo-vest head ring for replantation of total scalp avulsion can effectively improve the survival rate and survival area.

Anastomosis, Surgical ; methods ; Bandages ; Graft Survival ; Humans ; Lacerations ; surgery ; Necrosis ; etiology ; Prostheses and Implants ; Replantation ; methods ; Scalp ; injuries ; pathology ; surgery ; Skin Transplantation ; methods ; Surgical Flaps