Recent studies have shown that the physiological homeostasis of oral mucosal cells is regulated by the circadian clock.Dis-ruption or dysfunction of the circadian clock is closely associated with the development of oral squamous cell carcinoma(OSCC).Research based on the circadian clock offers a novel perspective on the pathogenesis and therapeutic strategies for OSCC.However,there is current-ly limited research on this topic,and people generally have insufficient understanding and recognition of the circadian clock.Given the complexity and challenges of circadian clock which is the fourth dimension of medical research,we organize relevant experts based on summarizing the current research results of circadian clock in the pathogenesis and precision diagnosis and treatment of OSCC,combining the scientific principles of the circadian clock's role and their long-term research experience,then summarizes and recommends the con-sensus opinions for the research of circadian clock in the pathogenesis mechanism and precision diagnosis and treatment of human OSCC,with the hope of providing guidance for the basic research and clinical application of circadian clock or circadian rhythm in the pathogene-sis mechanism and precision diagnosis and treatment of oral and maxillofacial squamous cell carcinoma.

Recent studies have shown that the physiological homeostasis of oral mucosal cells is regulated by the circadian clock.Dis-ruption or dysfunction of the circadian clock is closely associated with the development of oral squamous cell carcinoma(OSCC).Research based on the circadian clock offers a novel perspective on the pathogenesis and therapeutic strategies for OSCC.However,there is current-ly limited research on this topic,and people generally have insufficient understanding and recognition of the circadian clock.Given the complexity and challenges of circadian clock which is the fourth dimension of medical research,we organize relevant experts based on summarizing the current research results of circadian clock in the pathogenesis and precision diagnosis and treatment of OSCC,combining the scientific principles of the circadian clock's role and their long-term research experience,then summarizes and recommends the con-sensus opinions for the research of circadian clock in the pathogenesis mechanism and precision diagnosis and treatment of human OSCC,with the hope of providing guidance for the basic research and clinical application of circadian clock or circadian rhythm in the pathogene-sis mechanism and precision diagnosis and treatment of oral and maxillofacial squamous cell carcinoma.

Pseudorabies virus(PRV)is the etiological agent of pseudorabies in pigs,which is char-acterized by dyspnea,reproductive disorders,and neurological diseases,and it spreads widely a-round the world.Since 2011,the newly emerged PRV variants have resulted in poor immunity pro-tection of traditional vaccine strains,and the original method of vaccine strain preparation is time-consuming and labor-intensive.Therefore,it is urgently needed to develop an efficient screening method of the vaccine strain at present.Using CRISPR/Cas9 gene editing technology in this study,two single guide RNAs(sgRNA)were designed targeting the virulence gene TK of PRV variant strain CH/JX/2016,and then the enhanced green fluorescent protein the reporter(EGFP)gene was inserted at the TK locus by a homologous repair plasmid.After multiple rounds of plaque puri-fication,the rPRV-ΔTK/EGFP strain was obtained.The results showed the cleavage efficiency of the two sgRNAs was extremely high.The preparation of rPRV-ΔTK/EGFP strain was succeed af-ter only three rounds of purification,and the EGFP expressed normally.The CRISPR/Cas9 system can edit the PRV gene simply,rapidly,and efficiently,and exhibits great potential in the construction of vaccine candidate strains.Meanwhile,the rescued rPRV-ΔTK/EGFP strain not only could be used as a tracer strain in PRV variant infection progresses,but also for subsequent antivi-ral drug screening.

Porcine deltacoronavirus(PDCoV)is one of the important pathogens associated with swine viral diarrhea,which results in enormous economic losses in the pig industry.Among the known encoding proteins,the nucleocapsid(N)protein is relatively conserved and serves as the dominant antigen of coronaviruses,making it an ideal candidate for the early and accurate diagnosis of infection.In this study,the PDCoV N protein was induced and expressed in a prokaryotic system using IPTG treatment at low temperatures.Six-week-old BALB/c mice were immunized with the recombinant PDCoV N protein purified by nickel column.Isolated spleen cells were harvested and fused with SP2/0 cells once the serum titers reached 1∶10 000,as determined by indirect ELISA.The positive monoclonal hybridoma with the highest titer of secretory antibodies were screened af-ter three rounds of limited dilution.Ultimately,indirect immunofluorescent assay(IFA)and West-ern blot were employed to confirm the characterization of the antibodies,and the subtypes of the heavy and light chains of the antibodies were determined.One strain of hybridoma against the PD-CoV N protein,designated as 2G12,was obtained,with a titer approaching 1∶40 960.The IFA as-say showed PDCoV was specifically bound to 2G12.Furthermore,the Western blot assay further showed PDCoV in the supernatant of infected cells or recombinant PDCoV N protein,reacted well with 2G12.The subtype of MAbs was determined as IgG1 with the light chains being κ.The MAbs generated against the PDCoV N protein in this study provide valuable support for the development of a diagnostic kit for PDCoV and play an important role in the functional research of the PDCoV N protein.

Porcine deltacoronavirus(PDCoV)is one of the important pathogens associated with swine viral diarrhea,which results in enormous economic losses in the pig industry.Among the known encoding proteins,the nucleocapsid(N)protein is relatively conserved and serves as the dominant antigen of coronaviruses,making it an ideal candidate for the early and accurate diagnosis of infection.In this study,the PDCoV N protein was induced and expressed in a prokaryotic system using IPTG treatment at low temperatures.Six-week-old BALB/c mice were immunized with the recombinant PDCoV N protein purified by nickel column.Isolated spleen cells were harvested and fused with SP2/0 cells once the serum titers reached 1∶10 000,as determined by indirect ELISA.The positive monoclonal hybridoma with the highest titer of secretory antibodies were screened af-ter three rounds of limited dilution.Ultimately,indirect immunofluorescent assay(IFA)and West-ern blot were employed to confirm the characterization of the antibodies,and the subtypes of the heavy and light chains of the antibodies were determined.One strain of hybridoma against the PD-CoV N protein,designated as 2G12,was obtained,with a titer approaching 1∶40 960.The IFA as-say showed PDCoV was specifically bound to 2G12.Furthermore,the Western blot assay further showed PDCoV in the supernatant of infected cells or recombinant PDCoV N protein,reacted well with 2G12.The subtype of MAbs was determined as IgG1 with the light chains being κ.The MAbs generated against the PDCoV N protein in this study provide valuable support for the development of a diagnostic kit for PDCoV and play an important role in the functional research of the PDCoV N protein.

Objective:To comparatively evaluate the accuracy and the scan time of three full-arch scan strategies on the head-simulator, to explore a full-arch scan strategy with better clinical operability and high accuracy.Methods:A cross-controlled study design was used. A model with melamine-formaldehyde resin teeth and silica gel gingiva of an upper dental arch which can be fixed on a head simulator was scanned with an optical scanner (ATOS Core) in order to obtain the standard tessellation language (STL) dataset as reference. Intraoral scans were performed on the model fixed on the head simulator with four intraoral scanners (IOS) [A (TRIOS 3), B (CS 3600), C (CEREC Omnicam), D (iTero)]. The STL datasets were obtained from each of the four different IOS systems by using three scan strategies (scan strategies 1, 2 and 3 were composed of 10, 5 and 7 paths respectively) all by one attending doctor with 3 years of intraoral scanning experience. For each scanner and each scan strategy, nine scans were acquired. And the scan time was recorded for each scan. Following the scan strategy, the scan path was completed to obtain a full-arch digital model, and the scan time was recorded as full-arch scan time. Complementary scans were performed to fill the missing image, and this scan time was recorded as complementary scan time. The total scan time was obtained by adding full-arch scan time and complementary scan time. Through the Geomagic Wrap software, the three-dimensional (3D) models were overlaid by best fit alignment function and compared to obtain the root mean square values of the discrepancies by 3D compare function. The intraoral scanning datasets were compared with the reference for trueness. The nine intraoral scanning datasets were cross compared with same scan strategy and same intraoral scanner for precision.Results:There were no significant differences among the three scan strategies for trueness ( P>0.05), while the differences among the three scan strategies for precision were affected by difference IOSs ( P<0.05), and only scan strategy 3 showed the highest precision with all the four IOS. The full-arch scan time of scan strategies 1, 2 and 3 were (130±24), (72±17) and (90±19) s respectively ( P<0.05). For complementary scan time, scan strategy 2 [(50±24) s] took longer time than scan strategy 1 [(26±18) s] and scan strategy [(25±21) s] ( P<0.05), while no significant differences between the latter two ( P>0.05). For total scan time, scan strategy 1 [(156±31) s] took longer time than scan strategy 2 [(122±30) s ] and scan strategy 3 [(115±29) s ] ( P<0.05), while no significant differences between the latter two ( P>0.05). Conclusions:Full-arch scanning on the head-simulator with scan strategy 3 which can obtain scanning datasets with high accuracy, was more convenient to operate and took shorter scan time, and is generally suitable for intraoral scanners commonly used in clinic.

Objective: To compare the efficacy of goal-activity-motor environment (GAME) therapy and neurodevelopmental therapy (NDT) in the early intervention of high-risk infants with cerebral palsy (IHRCP), and to provide scientific evidence-based medical basis for early intervention of IHRCP. Methods: A total of 62 cases of IHRCP were enrolled in the Children′s Neurological Rehabilitation Center of the First Affiliated Hospital of Anhui Medi-cal University from June 2017 to December 2018.They were divided into GAME group (32 cases) and NDT group (30 cases) according to the admission order.Gross Motor Function Scale (GMFM), Fine Motor Function Measure (FMFM) and Gesell Development Scale (GDS) were used for detection and comparison.The differences among the gross motor, the fine motor score and the developmental quotient (DQ) between two groups before treatment, 9 months after treatment and 12 months after treatment, and the normalization rate and the incidence of cerebral palsy between the two groups at 12 months of age were compared. Results: (1) Motor function was as follows: at 9 months[GAME: (32.63±15.83) scores, (30.03±15.88) scores], [NDT: (33.37±15.61) scores, (29.67±12.54) scores] and at 12 months[GAME: (40.56±15.79) scores, (36.31±14.98) scores], [NDT: (40.47±15.50) scores, (36.73±14.58) scores] after treatment, and GMFM and FMFM scores in GAME and NDT groups were significantly higher than those before treatment[GAME: (27.56±14.24) scores, (21.75±11.35) scores], [NDT: (26.93±14.96) scores, (21.30±10.67) scores], and the differences were significant (all P<0.01), but there was no significant difference between the 2 groups (all P> 0.05). (2) DQ had no significant difference in DQ between GAME group(63.59±10.83) and NDT group (61.59±7.96) before treatment (P>0.05). The total DQ at 9 months, 12 months, the total DQ of GAME group (73.67±12.00, 81.59±13.03) was significantly higher than that of NDT group (66.05±9.54, 75.17±1.92) (all P<0.05). Among them, the improvement of GAME in speech (79.84±16.56, 83.19±17.05) at 9 months and 12 months, and adaptive ability(78.63±16.37, 85.78±13.60) were significantly higher than that of NDT group(71.63±13.36, 72.53±12.77), (68.20±14.97, 77.43±12.10), and the differences were significant (all P<0.05). (3) Prognosis was as follows: at 12 months after treatment, 25 cases in GAME group and 23 cases in NDT group developed into normal children, there was no significant difference in the normalization rate between the 2 groups (P>0.05); the incidence of cerebral palsy was present in 6 cases in GAME group and 5 cases in NDT group, and there was no significant difference between the 2 groups (P>0.05). Conclusions GAME therapy and NDT had significant effects on both gross and fine exercise of IHRCP, and the efficacy of the two methods is similar.Both GAME therapy and NDT can equally promote IHRCP development into normal infants and reduce the incidence of cerebral palsy.

Objective To investigate the accuracy of transthoracic echocardiography (TTE) for diagnosing left cardiac valve calcification in elderly patients.Methods In this retrospective study,410 elderly patients died in Beijing Hospital from 1982 to 2015 were collected.They all had full pathological cardiac valve examination data in the autopsy examination and full mortem TTE data.They were classified into three groups according to the year of death:group 1982-1995,group 1996-2005 and group 2006-2015,as well as into three groups according to the age of death:group 60-75,group76-90 and group 91-106.Results The sensitivity,specificity,diagnostic coincidence rate,positive likelihood ratio (PLR),negative likelihood ratio (NLR),positive predictive value (PPV) and negative predictive value(NPV) of TTE versus autopsy pathological findings as golden standard for diagnosing left cardiac valve calcification were 88.6％,28.2％,43.7％,1.234,0.405,0.298 and 0.878.Compared with group 1982-1995,the group 1996-2005 and group 2006-2015 showed the sensitivity and PPV of TTE versus golden standard autopsy findings for diagnosing left cardiac valve calcification in the elderly were increased while the specificity,PLR,NLR and NPV were decreased.Compared with group 60-75,the group 76-90 and group 91-106 showed that the sensitivity and PPV of TTE versus golden standard autopsy findings for diagnosing left cardiac valve calcification were increased,while the specificity and diagnostic coincidence rate were decreased.The sensitivity,specificity,diagnostic coincidence rate,PLR,NLR,PPV,NPV of TTE versus golden standard autopsy findings for the diagnosis of aortic valve calcification and mitral valve calcification in the elderly were 88.2％ and 44.0％,30.8％ and 75.3％,42.7％ and 71.5％,1.275 and 1.780,0.382 and 0.744,0.250 and 0.198,0.909 and 0.906,respectively.Conclusions TTE provides high sensitivity for diagnosing left cardiac valve calcification in the elderly,especially for diagnosing the aortic valve calcification,but the specificity is not satisfactory.

Serum Cholyglycine (CG)is a main component of human bile acid,one of the conjuga-ted bile acids formed by the combination of bile acid and glycine is synthesized in the liver cells.Glycochol-ic acid (CG)as a clinical indicator for detecting hepatobiliary disorders,and the traditional index of liver
function test compared has greater advantages,its detection for intrahepatic cholestasis,liver disease and biliary system diseases diagnosis,treatment and prognosis analysis of pregnancy to provide an important ba-sis.

RNA interference (RNAi) is a powerful endogenous process initiated by short double stranded RNAs, which results in sequence-specific posttranscriptional gene silencing. Because any protein that causes or contributes to a disease is susceptible to RNAi, the RNAi has high potential for therapeutic treatments. In a clinical setting, however, there are many obstacles to targeted delivery of small interfering RNA (siRNA) in vivo, specificity and stability of the RNAi reagents. In this review, we focus on recent progress in the development of efficient siRNA delivery vehicles to help the application of siRNA to in vivo therapy.
Drug Carriers ; Gene Transfer Techniques ; Humans ; Molecular Targeted Therapy ; methods ; trends ; RNA Interference ; RNA, Small Interfering ; administration & dosage ; genetics ; metabolism ; Transfection