Objective:To explore the efficient construction of HSF1 gene knockout mouse model using CRISPR/Cas9 gene editing technology, and to establish the early basis for the mouse model of primary osteosarcoma.Methods:According to exon 9 of HSF1 gene structure, the corresponding GRNA (guideRNA) was selected and screened. Then the transcription template of sgRNA (small guide RNA) was amplified by PCR, and four up stream primers were obtained. Subsequently, sgRNA was transcribed in vitro and screened by Tube Screen platform to screen the sgRNA with effective cutting, and the sgRNA with the highest cutting efficiency was selected from the screening results for subsequent experiments. The transcription template of SPCas9mRNA was amplified by PCR, and then Cas9mRNA was transcribed in vitro. The sgRNA transcribed in vitro and Cas9mRNA were injected into the fertilized eggs of healthy C57BL/6 mice, and the tissue was extracted from the tail of the born mice and identified by PCR sequencing. Heterozygous female mice of F0 generation were selected to mate with wild-type male mice too btain F1 generation off spring. The mutation of gene bases of F1 generation mice was detected by AGAR gel electrophoresis and gene sequencing. The heterozygous male mice of the F1 generation and female mice of the F0 generation were back crossed to obtain the F2 generation daughter mice. The tail tissues were cut and sequenced to obtain the F2 generation homozygous knockout mice. PCR was used to observe the cutting efficiency of sgRNA and the sequencing of rat tail tissue, and SNAPGene software was used for gene sequence alignment to determine the deletion of base fragments.Results:The up stream primers sgRNA-1 Primer-f, sgRNA-2 Primer-f, sgRNA-3 Primer-f, sgRNA-4 Primer-f and down stream primers sgRNA-4 Primer -r were obtained by PCR amplification. After in vitro tran scription and screening of sgrRNA, sgrRNA-1, sgrRNA-2 and sgrRNA-4 had high cleavage efficiency and were selected for subsequent experiments. T7 promoter was added to the 5 'end of Cas9 mRNA, and Cas9 mRNA was obtained by PCR and in vitro transcription kit. Mixed Cas9-sgRNA solution was injected into the fertilized eggs of mice and cultured. The cultured two-cell fertilized eggs were injected into the ampulla of the pseudo pregnant female mice, and the F0 generation mice were obtained successfully. A total of 8 heterozygous mice of F0 generation were obtained by Agar gel electrophoresis. Three heterozygous knockout mice of F1 generation were obtained by breeding the female heterozygous mice of F0 generation with healthy wild-type male mice and PCR and sequencing. Three heterozygous male mice of F1 generation were back crossed with female mice of F0 generation 3 to obtain F2 generation mice. Through the observation of electrophoresis and sequencing results of F2 generation mice, it was confirmed that 7 mice were missing HSF1 base sequence, and the electrophoresis results showed mutant bands and no wild-type bands, which were identified as homozygous. The F2 generation homozygous mice were able to breed stably. As eries of results proved that the HSF1 gene knockout mouse model was successfully established in this experiment.Conclusion:CRISPR/Cas9 technology was successfully used to construct HSF1 gene knockout mouse model, with strong stability and high reproducibility, which laida foundation for further study of HSF1 gene expression products and establishment of mouse model of primary osteosarcoma.

Objective To investigate the positive practice environments ( PPE ) of nurses and influencing factors at public hospitals , for reference of building a better PPE .Methods A national cross-sectional survey was performed at 77 public hospitals across seven provinces/metropolises, involving 5374 nurses.PPE included organizational management (internal) and nurses-patient relationship (external). Results The scoring of positive practice environment was 18.51 ±4.69 (total score of 40).The scoring of organizational management and nurses-patient relationship was 9.87 ±3.11 and 8.64 ±2.51 respectively. The scoring of PPE of nurses of general hospital ( GH) was higher than that of traditional Chinese medicine hospital(TCMH) (18.68 ±4.68 versus 18.08 ±4.67, P<0.01).Multivariable analysis showed that , compared with nurses who had not very much pressure about performance assessment , the scores of those who had were declined (βGH =-1.15, 95％CI: -1.55 to -0.76;βTCMH =-1.29, 95％CI: -1.92 to-0.66 ) );compared with nurses who paid less efforts in communicating with their patients , the scoring of those with greater efforts was higher (βGH =2.43, 95％CI:2.00 to 2.86;βTCMH =2.84, 95％CI:2.19 to 3.49).Conclusions PPE of nurses is poor in general in China , and the externally stressful nurse-patient relationship deserves greater attention and efforts than inefficient organization management internally .To improve PPE of nurses , hospitals need to moderate nurses′performance assessment and the nurses need to pay more attention to patient communication .

With the further increase of the global cancer burden, various cancers are increasingly challenging human health status and quality of life. Thus, early screening of cancer is crucial. Urinary volatile organic compounds (VOCs) detection techniques have the advantages of easy access to samples, high acceptance of patients, non-invasive, and so on, which have been favored and concerned by researchers. In this paper, existing techniques and methods for cancer diagnosis based on urine VOCs were described, relevant studies on the use of urine VOCs for cancer diagnosis were reviewed, and the barriers and future perspectives of the technique were discussed. This paper can be a reference for researchers working in the direction of urinary VOCs detection, a multidisciplinary field that spans medicine and materials science.

Objective: To investigate the role of JAZF1 gene rearrangement in the diagnosis and differential diagnosis of endometrial stromal sarcomas by fluorescence in situ hybridization (FISH). Methods: JAZF1 gene rearrangement was analyzed by FISH in 129 cases of ESS diagnosed from January 2008 to December 2016 including 105 cases of low-grade endometrial stromal sarcoma (LG-ESS), 21 cases of high-grade endometrial stromal sarcoma (HG-ESS) and 3 cases of undifferentiated uterine sarcoma (UUS). Sixteen cases of the related tumours in uterus were also collected as control group. The results were compared with our previous studies of JAZF1/JJAZ1 fusion gene in ESS by RT-PCR. Results: Detection of JAZF1 gene rearrangement by FISH was successfully analyzed in 144 cases. JAZF1 gene alteration was detected in 63 cases, all of which were LG-ESS, with an overall positivity of 60.6% (63/104), while no JAZF1 gene rearrangement was found in all other cases. JAZF1 gene rearrangement was present in LG-ESS with classic histology (69.3%, 52/75), smooth muscle differentiation (2/10), sex cord-like differentiation (4/5), fibromyxoid change (1/5), clear cell change (0/1), skeletal muscle differentiation (0/1), and schwannoma-like palisading pattern (0/1). The different components in all the cases of LG-ESS with variant histology had the clonal origin, with or without JAZF1 gene alteration. Compared to the results of JAZF1/JJAZ1 fusion gene by RT-PCR, the positive rate of JAZF1 gene rearrangement in LG-ESS by FISH (61.9%, 26/42) was significantly higher than that of RT-PCR (30.0%, 12/40; P<0.01). Conclusions JAZF1 gene rearrangement is present only in LG-ESS, but not in HG-ESS, UUS or other related tumours in uterus. The frequency of JAZF1 gene rearrangement varies between classic LG-ESS and different morphologic variants. It is frequently, but not consistently, present in classic LG-ESS and less often positive in variant cases. The results of JAZF1 gene alterations in LG-ESS with different morphologic variants support the contention that the endometrial stromal and their variant morphologic components have the same clonal origin. Detection of JAZF1 gene rearrangement by FISH is very useful for the diagnosis and differential diagnosis of ESS.

Objective: To investigate craniofacial differences in individuals with hypodontia and explore the relationship between craniofacial features and the number of congenitally missing teeth. Methods: A cross-sectional study was conducted among 261 Chinese patients (males, 124; females, 137; age, 7–24 years), divided into four groups (without hypodontia: no teeth missing, mild: one or two missing teeth, moderate: three to five missing teeth, severe: six or more missing teeth) according to the number of congenitally missing teeth. Differences in cephalometric measurements among the groups were analyzed. Further, multivariate linear regression and smooth curve fitting were performed to evaluate the relationship between the number of congenitally missing teeth and the cephalometric measurements. Results: In patients with hypodontia, SNA, NA-AP, FH-NA, ANB, Wits, ANS-Me/N-Me, GoGn-SN, UL-EP, and LL-EP significantly decreased, while Pog-NB, AB-NP, N-ANS, and S-Go/N-Me significantly increased. In multivariate linear regression analysis, SNB, Pog-NB, and S-Go/N-Me were positively related to the number of congenitally missing teeth. In contrast, NA-AP, FH-NA, ANB, Wits, N-Me, ANS-Me, ANS-Me/N-Me, GoGn-SN, SGn-FH (Y-axis), UL-EP, and LL-EP were negatively related, with absolute values of regression coefficients ranging from 0.147 to 0.357. Further, NA-AP, Pog-NB, S-Go/N-Me, and GoGn-SN showed the same tendency in both sexes, whereas UL-EP and LL-EP were different. Conclusions Compared with controls, patients with hypodontia tend toward a Class III skeletal relationship, reduced lower anterior face height, flatter mandibular plane, and more retrusive lips. The number of congenitally missing teeth had a greater effect on certain characteristics of craniofacial morphology in males than in females.

Objective:To retrospectively identify the risk factors for postoperative nausea and vomiting (PONV)in the obese patients undergoing laparoscopic sleeve gastrectomy(LSG).Methods:The medical records from the obese patients who underwent elective laparoscopic sleeve gastrectomy from January 2018 to July 2022 were retrospectively collected. PONV was defined according to the use of remedial antiemetics in the nursing record sheet, and the patients were divided into PONV group and non-PONV group according to the occurrence of PONV that required treatment. The logistic regression analysis was used to identify the risk factors for PONV after LSG.Results:A total of 1 264 obese patients were included in this study, and there were 263 patients in PONV group, and the incidence of PONV was 20.81%. According to the results of multifactorial logistic regression analysis, female( OR=1.533, 95% CI 1.007-2.334, P=0.046), higher level of serum alanine aminotransferase concentrations ( OR=1.006, 95% CI 1.002-1.009, P=0.001), higher level of C-reactive protein ( OR=1.013, 95% CI 1.005-1.022, P=0.001), general anesthesia combined with nerve block (general anesthesia combined with TAPB: OR=2.737, 95% CI 1.817-4.121, P<0.001; general anesthesia combined with other nerve block: OR=1.899, 95% CI 1.249-2.889, P=0.003) and intraoperative use of sufentanil ( OR=2.114, 95% CI 1.308-3.415, P=0.002) were independent risk factors for PONV( P<0.05). However, the higher level of serum follicle-stimulating hormone concentrations ( OR=0.941, 95% CI 0.895-0.988, P=0.015), intraoperative use of dexmedetomidine ( OR=0.640, 95% CI 0.417-0.982, P=0.041), and administration of prophylactic antiemetic medication (antiemetic drugs during operation OR=0.669, 95% CI 0.469-0.955, P=0.027; antiemetic drugs after operation OR=0.303, 95% CI 0.182-0.503, P<0.001; antiemetic drugs during and after operation OR=0.215, 95% CI 0.107-0.434, P<0.001) were protective factors for PONV. Conclusions:Female, higher levels of serum alanine aminotransferase and C-reactive protein, general anesthesia combined with nerve block and intraoperative use of sufentanil are independent risk factors for PONV, while higher levels of follicle-stimulating hormone, intraoperative use of dexmedetomidine and administration of prophylactic antiemetic medication are protective factors for PONV among obese patients undergoing LSG.

The latest epidemiological data suggests that the situation of adult diabetes in China is severe, and metabolic diseases have become significant chronic illnesses that have a serious impact on public health and social development. After more than six years of practice, the National Metabolic Management Center(MMC) has developed distinctive approaches to manage metabolic patients and has achieved a series of positive outcomes, continuously advancing the standardized diagnosis and treatment model. In order to further improve the efficiency, based on the first edition, the second edition guideline was composed by incorporating experience of the past six years in conjunction with the latest international and domestic guidelines.

Multi-modal therapeutics are emerging for simultaneous diagnosis and treatment of cancer. Polymeric carriers are often employed for loading multiple drugs due to their versatility and controlled release of these drugs in response to a tumor specific microenvironment. A theranostic nanomedicine was designed and prepared by complexing a small gadolinium chelate, conjugating a chemotherapeutic drug PTX through a cathepsin B-responsive linker and covalently bonding a fluorescent probe pheophorbide a (Ppa) with a branched glycopolymer. The branched prodrug-based nanosystem was degradable in the tumor microenvironment with overexpressed cathepsin B, and PTX was simultaneously released to exert its therapeutic effect. The theranostic nanomedicine, branched glycopolymer-PTX-DOTA-Gd, had an extended circulation time, enhanced accumulation in tumors, and excellent biocompatibility with significantly reduced gadolinium ion (Gd