Objective To explore the effects of cyclophosphamide administered by different routes or in different doses on the embryo-fetal development in pregnant rabbits, and to determine the optimal mode of cyclophosphamide administration to induce fetal malformation.Methods Pregnant rabbits were divided into control group C (saline), group Y1 (intravenous injection of 15 mg/kg cyclophosphamide,), group Y2 (subcutaneous injection of low dose cyclophosphamide, 20 mg/kg), and group Y3 (subcutaneous injection of high dose cyclophosphamide, 30 mg/kg).Each rat was administrated according to the corresponding mode once daily on GD10~13.The day of conception was designated as GD0.The pregnant rabbits were sacrificed and dissected on GD28.Then, the number of corpora lutea and implantation, the weight of uterus with contained fetus, and fetal resorption rate were detected, the fetuses were removed and the fetal sex, body length, tail length, the number of live births and stillbirths were recorded, and the appearance of deformities, visceral deformities and skeletal malformations were detected.Results Pregnant rabbit fetuses in the cyclophosphamide intravenous injection group and subcutaneous injection of low dose cyclophosphamide group showed deformities.The appearance malformation rates in the two groups were 30.77% and 95.65%, the skeletal deformity rates were 7.69% and 73.91%, and the visceral abnormality rates were 20.51% and 47.83%, respectively.The fetal resorption rate in the high dose cyclophosphamide subcutaneous injection group was 100%.Conclusions Subcutaneous injection of 20 mg/kg cyclophosphamide to pregnant rabbits at GD10~13 can be used as a positive administrationmethod for rabbit embryo-fetal developmental toxicity test.Thismethod has the advantages of short administration period, easy operation, few fetus resorption, and high rate of fetal malformation, thus, providing the evidence for selection of appropriate model of rabbit embryo-fetal developmental toxicity.

OBJECTIVE To investigate the reasons and preventive measures of recurrent laryngeal nerve injury during operation in patients with benign thyroid neoplasm. METHODS The clinical data of 2243 cases with benign thyroid neoplasm were reviewed. The relations among different neoplasm regions, different procedures and injury of recurrent laryngeal nerve were analyzed. RESULTS Injury incidence of recurrent laryngeal nerve in this study was 3.0 %. In cases with nodular goiter on the dorsum of thyroid and recurrent laryngeal nerve exposed during operation, injury incidence of recurrent laryngeal nerve was 1.0 %. And there was no recurrent laryngeal nerve injury in cases with nodular goiter on other region of the thyroid. In cases used locality protection during operation, injury incidence of recurrent laryngeal nerve was 7.6 % when benign lesions on the dorsum of thyroid and was 0.17 % when benign lesions on other region of the thyroid. CONCLUSION In cases with nodular goiter or adenoma on the dorsum of thyroid, exposing recurrent laryngeal nerve is necessary during operation and in cases with benign lesions on other region of thyroid, locality protection is useful. There are three major routes for recurrent laryngeal nerve exposure.

Objective To investigate the plasma vitamin D3 level and its association with B cell subsets of patients with primary Sj(o)gren's syndrome (pSS).The role of vitamin D3 levels in the pathogenesis of pSS was explored.Methods The expression of plasma vitamin D3 levels of 55 patients with pSS and 32 controls were analyzed by ELISA.Frequencies of peripheral blood CD19+CD27-na(i)ve B cells,CD19+CD27+ memory B cells and CD19+CD27high plasma cells were analyzed by flow cytometry in 34 pSS patients without therapy and 22 controls.The relationship between the vitamin D3 levels and B cell subsets,SSDAI,tear flow rate,saliva flow rate,rheumatoid factor,immunoglobulin was analyzed in pSS patients.Non-parametric test,t test,one-way ANOVA,x2test,Pearman's and Spearman's correlation analysis were used for statistical analysis.Results ① There was significant difference in the levels of plasma 1,25 (OH)2D3 between the pSS patients group and normal control group,1,25 (OH)2D3 was significantly lower in pSS patients than that in the normal control group [24.17(22.20,28.41) pg/ml and 41.25(23.38,62.18) pg/ml,P＜0.05],and that was also obviously lower in the active group [22.64(20.74,24.90) pg/ml] than that in the normal control group (P＜0.05),and that was also obviously lower in the active disease group than that in the inactive disease group [25.39 (23.16,33.09) pg/ml,P＜0.05],but there was no difference between the inactive group and the normal control group (P＞0.05).② The percentage of peripheral blood of CD19+CD27high plasma cells and CD19+CD27+ memory B cells in CD19+ cells was reduced in patients in the pSS group compared with the control group [(0.89±0.30)％ and (1.72±0.43)％,(24±8)％ and (34±5)％; P＜0.05],and that was also significantly lower in the active group [(1.03±0.59) ％ ; (26± 10)％] and inactive group [(1.00±0.16)％,(26± 3)％] than that in the normal controls (P＜0.05).However,there was no difference between the active group and the inactive group (P＞0.05),but the frequency of peripheral blood of CD19+ CD27-naive B cells in CD19+ B cells was increased in patients with pSS compared with normal control group [(75.4±7.5)％ and (63.9±5.2)％,P＜0.05],and that was also significantly higher in the active group [(73.4±9.7)％] and inactive group [(73.3±2.9)％] than that in the normal control (P＜0.05),there was no difference between the active group and the inactive group.③ Significant negative correlation was observed between 1,25 (OH)2D3 and the percentage of peripheral blood CD19+CD27+ memory B cells in CD19+ cells as well as immunoglobulin G(r=-0.627,P=0.039; r=-0.657,P＜0.01) level.Conclusions These results demonstrate that abnormality of vitamin D levels may play an important role in the pathogenesis of pSS.

Objective: To evaluate the effect and mechanism of down-regulating the expression of REV7 on the radiosensitivity of human colon cancer cell HCT116. Methods: HCT116 cells were cultured and the expression of REV7 was down-regulated by RNA interference technique. HCT116 cells were divided into the blank group, negative control transfected with negative RNA oligo group and REV7 expression down-regulation transfected with REV7 RNA oligo group, respectively. The cell proliferation was determined by colony formation assay. The expression levels of the proteins of relevant genes were detected by Western blot. The level of cell apoptosis and non-homologous end joining was evaluated. Results: The colony formation rate was significantly reduced in THE REV7 siRNA group after 6Gy irradiation (P<0.05). The down-regulating efficiency rate of REV7 gene was>60% in the REV7 siRNA group. The expression levels of γH₂AX and Caspase9 were significantly up-regulated, whereas those of KU80 and XRCC4 were remarkably down-regulated in the REV7 siRNA group (all P<0.05). Conclusions The radiosensitivity of human colon cancer cell HCT116 can be increased by down-regulating the expression of REV7. The underlying mechanism may be related to the lower incidence rate of non-homologous end joining.

Objective To investigate the efficacy of CT scanning for 3D reconstruction and 3D printing in the internal fixation for in situ femoral condyle fractures.Methods A retrospective study was conducted of the 111 patients with in situ femoral condyle fracture who had been treated by internal fixation at Department of Traumatic Orthopaedics,The First People's Hospital of Hefei from June 2013 to January 2018.They were 59 males and 52 females,aged from 20 to 59 years(average,47 years).In 74 of them (observation group),the internal fixation was assisted by 3D printing using 3D images.There were 38 males and 36 females with an age of 44.5 ±4.7 years;there were 20 cases of type C1,40 cases of type C2 and 14 cases of type C3 according to the AO classification.In the other 37 patients (control group),the operation was assisted only by 3 D scanning.There were 21 males and 16 females with an age of 43.1 ±4.6 year;there were 8 cases of type C1,19 cases of type C2 and 10 cases of type C3 according to the AO classification.The 2 groups were compared in terms of operation time,clinical fracture healing time,intraoperative blood loss,hospital stay,number of fluoroscopy,knee joint score of Hospital for Special Surgery (HSS),fixation modes and postoperative complications.Results There were no significant differences in the preoperative general data between the 2 groups,showing comparability (P ＞ 0.05).All the patients were followed up for 8 to 12 months (average,10.6 months).The observation group had significantly shorter operation time (62.3 ± 4.7 minutes),fracture healing time (21.2 ± 3.4 weeks) and hospital stay (14.9 ± 3.3 days) than the control group (89.5 ± 6.0 minutes,25.1 ± 3.5 weeks,23.5 ± 3.5 days) (P ＜ 0.05).In the observation group,the intraoperative blood loss (84.1 ± 11.1 mL),fluoroscopy number (2.1 ±0.3 times) and HSS score (94.1 ± 4.2 points) were significantly less than those in the control group (129.6 ± 14.7 mL,4.7 ± 0.4 times and 86.5 ± 4.6 points) (P ＜ 0.05).There were no significant differences between the 2 groups in the use of L type iliac plate,dynamic iliac screws,retrograde interlocking intramedullary nail or minimally invasive internal fixation system (P ＞ 0.05).There were no significant differences in abnormal healing,joint adhesion,fixation fracture or incision infection between the 2 groups either (P ＞ 0.05).Conclusions CT scanning for 3D reconstruction can be used for multi-angle observation of the bone fragments in patients with femoral condyle fracture and 3D printing based on 3D reconstruction for manufacture of physical model of individualized fracture to assist the internal fixation.Combination of the two can help choose an appropriate fixation mode to facilitate fracture reduction.

Objective:To explore the effects of the γ-aminobutyric acid(GABA) neurons and melanin-concentrating hormone (MCH) neurons of the nucleus accumbens (NAc)-lateral hypothalamic area (LHA) neural pathway on the rewarding feeding(palatable food sweat condensed milk) in the obesity rats.Methods:Total 142 male Wistar rats of SPF grade were divided into normal diet (ND) group ( n=68) and high-fat diet induced obesity (DIO) group ( n=74) according to the principle of body mass matching. The rats in the two groups were given normal diet and high-fat diet for 8 weeks. Eight weeks later, 6 DIO rats were randomly selected to observe the nerve projection from GABA neurons in NAc to MCH neurons in LHA by fluorogold retrograde tracing combined fluorescence immunohistochemistry. And the expressions of c-Fos and MCH in LHA after ingestion of sweet condensed milk(rewarding feeding) were observed by fluorescence immunohistochemistry (6 rats in each group). GABA receptor agonist Musimol or GABA receptor antagonist Bicuculine was microinjected into the nucleus of LHA to observe the effect of GABA on rewarding food intake in ND and DIO rats ( n=8 in each group), and the changes of rewarding food intake after blocking MCH signal ( n=8 in each group). SPSS 17.0 was used for statistical analysis, two-way ANOVA and post hoc Bonferroni test were used for comparison among multiple groups, and t-test was used for comparison between two groups. Results:After 8 weeks of high-fat diet modeling, the intake of delicious food in DIO rats was significantly higher than that in ND rats((12.52±2.29) mL, (7.45±1.23) mL, t=4.778, P<0.01) after satiety.The results of fluorogold retrograde tracing combined with fluorescence immunohistochemistry showed that GABA neurons in NAc projected nerve fibers to neurons in LHA, and GABA A receptors in some neurons in LHA coexisted with MCH.The results of NAc-LHA pathway on delicious food intake showed that the interaction between rat group and drug intervention was significant( F=9.869, P<0.01). Simple effect analysis showed that the intake of delicious food after microinjection of Musimol into LHA nucleus of ND rats was significantly lower than that of microinjection normal saline ((4.25±1.38) mL, (7.29±1.49) mL, P<0.01), while the intake of delicious food after injection of Bicuculine was significantly higher than that of microinjection normal saline((10.72±2.11) mL, (7.29±1.49) mL, P<0.05). The intake of delicious food after microinjection of Musimol into LHA nucleus in DIO group was significantly lower than that of microinjection normal saline((3.51±1.77)mL, (13.68±2.95) mL, P<0.01), but there was no significant difference between microinjection Bicuculine and microinjection normal saline ((14.83±3.44) mL, (13.68±2.95) mL, P>0.05). The results of blocking MCH signal on delicious food intake showed that the interaction effect between SNAP-94847 and Bicuculine intervention was not significant ( F=1.468, P>0.05). The main effect of SNAP-94847 intervention was significant ( F=15.880, P<0.01)and the main effect of Bicuculine intervention was significant ( F=6.930, P<0.05). After intracerebroventricular injection of MCH receptor blocker SNAP-94847, the delicious food intake of ND rats was significantly less than that of injection normal saline((4.78±1.72) mL, (7.63±2.77) mL, P<0.05), and it was not affected by pre injection of Bicuculine in LHA ((6.24±2.18) mL, (4.78±1.72) mL, P>0.05). In the DIO rats, the interaction effect between SNAP-94847 and Bicuculine intervention was not significant( F=0.006, P>0.05). The main effect of SNAP-94847 intervention was significant ( F=18.46, P<0.01) and the main effect of Bicuculine intervention was not significant ( F=2.059, P>0.05). After intracerebroventricular injection of MCH receptor blocker SNAP-94847, the delicious food intake of DIO rats was significantly lower than that of injection normal saline((6.89±2.11) mL, (12.19±4.36) mL, P<0.05), and it was not affected by pre injection of Bicuculine in LHA ((8.72±2.26) mL, (6.89±2.11) mL, P>0.05). Conclusion:GABAergic signal in NAc can regulate the expression of MCH in neurons of LHA. In the DIO rats, the sensitivity of MCH neurons in LHA to satiety signal decreases and the hedonic feeding increases.

Objective To explore the effect of silencing Rev1 gene on the proliferation and apoptosis induction of human colon cancer cell line THC8307 after X-ray irradiation.Methods Rev1 siRNA was transferred into THC8307 cells and its transferring efficiency was tested by RT-PCR and Western blot assay.The cells were divided into blank control group,negative control group and radiation group.After 6 Gy irradiation,cell proliferation were detected by MTI assay,cell apoptosis were detected by flow cytometry (FCM),and the expressions of PCNA,γ-H2AX,P53,Bax,and Bcl-2 proteins in each group were detected by Western blot.Results Compared with the blank control group,after 6 Gy irradiation,the proliferation rate of the silenced Rev1 was reduced (t =7.53,P ＜ 0.05) and apoptosis rate was increased (t =6.23,P ＜ 0.05).The expression of PCNA and Bcl-2 decreased (t =4.39,6.13 P ＜0.05),and the expressions of γ-H2AX,P53 and Bax increased (t =5.48,5.09,3.32,P ＜ 0.05).Conclusions Silencing Rev1 gene inhibits the proliferation of colon cancer cells,promotes apoptosis,and increases cell radiosensitivity to X-rays.

Objective:To investigate the expression levels of serum calcitonin (CT) and osteoponin (OPN) in peripheral blood of patients with atrial fibrillation (AF) and their relationship with AF.Methods:A case control study was conducted, 160 AF patients treated in Shaanxi Provincial People's Hospital from June 2020 to December 2021 were selected as case group, and 160 healthy people in the same period were selected as control group. The expression levels of serum CT and OPN in the two groups were analyzed retrospectively, and the correlation between them and AF was analyzed. The value of CT and OPN levels in predicting the occurrence of AF was analyzed by receiver operating characteristic (ROC). Kappa consistency test was used to analyze the efficacy of the combination of them in predicting the occurrence of AF.Results:The serum calcitonin level ((13.5±3.2) ng/L) in the case group was lower than that in the control group ((17.3±3.1) ng/L), and the osteopontin level ((53.8±8.2) μg/L) was higher than that of the control group ((44.1±6.8) μg/L), the difference between the two groups was statistically significant ( t=10.79, P<0.001; t=11.50, P<0.001). The results of binary logistic regression analysis showed that serum calcitonin was the protective factor of atrial fibrillation ( OR=0.723, 95% CI: 0.661-0.790, P<0.001), and osteopontin was the risk factor ( OR=1.183, 95% CI: 1.131-1.237, P<0.001). ROC analysis showed that the area under curve (AUC) of CT, OPN and their combination in predicting AF were 0.794, 0.824, and 0.892, respectively. The predictive critical value for serum CT was <15.0 ng/L and >48.5 μg/L for OPN. The sensitivity, specificity of the combination in AF prediction were 67.50% and 93.75% respectively, and Kappa value was 0.613. Conclusion:The expression of serum CT and OPN was abnormal in patients with atrial fibrillation. The prediction of AF by combined examination of the two had a high degree of consistency with the actual results, which could provide reference for early diagnosis and treatment of AF. However, the rate of missed diagnosis was relatively high, which needs attention in clinical application.

Objective: To explore the relationship between processed food consumption and blood pressure level of students in a university in Yunnan Province, so as to provide the reference for preventing hypertension in university students. Methods: In October 2021, a cluster sampling method was used to select 4 781 freshmen from a university in Kunming, Yunnan Province. The frequency of processed food consumption of university students was assessed by using the dietary frequency questionnaire, and height, weight and blood pressure were measured. Mann-Whitney test and Kruskal-Wallis test were used to compare the differences in blood pressure level of university students with different demographic variables, and the association between processed food consumption and blood pressure level was analyzed with a generalized linear model. Results: Among the students of a university in Yunnan Province, the detection rates of systolic prehypertension and hypertension were 33.86% and 1.23%, and the detection rates of diastolic prehypertension were 32.13% and hypertension 7.22%. The results of generalized linear model analysis showed that after controlling for demographic variables and other variables that might affect the blood pressure level of university students, the consumption of processed food (bread and cake: β =0.15, 95% CI =0.01-0.29) and ultra processed food (coffee beverage: β =-0.29, 95% CI =-0.54--0.03) were associated with systolic blood pressure level( P <0.05). The consumption of processed food (salted duck egg: β =0.21, 95% CI =0.01-0.41) was correlated with the diastolic blood pressure of college students ( P <0.05). Conclusions Processed food consumption in university students may increase the risk of high blood pressure.The education of healthy eating among college students should be strengthened to reduce the consumption of processed foods.

The emergence of SARS-CoV-2 variants of concern and repeated outbreaks of coronavirus epidemics in the past two decades emphasize the need for next-generation pan-coronaviral therapeutics. Drugging the multi-functional papain-like protease (PLpro) domain of the viral nsp3 holds promise. However, none of the known coronavirus PLpro inhibitors has been shown to be in vivo active. Herein, we screened a structurally diverse library of 50,080 compounds for potential coronavirus PLpro inhibitors and identified a noncovalent lead inhibitor F0213 that has broad-spectrum anti-coronaviral activity, including against the Sarbecoviruses (SARS-CoV-1 and SARS-CoV-2), Merbecovirus (MERS-CoV), as well as the Alphacoronavirus (hCoV-229E and hCoV-OC43). Importantly, F0213 confers protection in both SARS-CoV-2-infected hamsters and MERS-CoV-infected human DPP4-knockin mice. F0213 possesses a dual therapeutic functionality that suppresses coronavirus replication via blocking viral polyprotein cleavage, as well as promoting antiviral immunity by antagonizing the PLpro deubiquitinase activity. Despite the significant difference of substrate recognition, mode of inhibition studies suggest that F0213 is a competitive inhibitor against SARS2-PLpro via binding with the 157K amino acid residue, whereas an allosteric inhibitor of MERS-PLpro interacting with its 271E position. Our proof-of-concept findings demonstrated that PLpro is a valid target for the development of broad-spectrum anti-coronavirus agents. The orally administered F0213 may serve as a promising lead compound for combating the ongoing COVID-19 pandemic and future coronavirus outbreaks.
Animals ; Coronavirus Papain-Like Proteases/antagonists & inhibitors* ; Cricetinae ; Humans ; Mice ; Pandemics ; SARS-CoV-2/enzymology* ; COVID-19 Drug Treatment