Objective To evaluate the methods and efficacy of microsurgical treatment for the premalignant and malignant lesions of the conjunctiva. Methods Forty-seven patients witn premalignant and malignant lesions of the conjuncitva were managed by microsurgical, these patients include 12 melanoma, 26 squamous cell carcinoma, 6 Bowen’ diseases, 3 primary acquired melanosis. The surgical method differs with limbal tumors, extralimbal tumors, and primary acquired melanosis. Results In all 47 patients, the tumor was completely removed in in one procedure. After follow-up for 4~60 months(mean 17 months), these were no tumor recurrence. Conclusion It is effective methods that premalignant and malignant lesions of the conjunctiva are managed by microsurgical resection, alcohol application, and supplemental cryotherapy. Careful propeirativa clinical evaluation of patient with a conjunctiva neoplasm is important in making the correct diagnosis and planning the surgical approach.

The accuracy and repeatability of computer aided cervical vertebra landmarking (CACVL) were investigated in cephalogram. 120 adolescents (60 boys, 60 girls) aged from 9.1 to 17.2 years old were randomly selected. Twenty-seven landmarks from the second to fifth cervical vertebrae on the lateral cephalogram were identified. In this study, the system of CACVL was developed and used to identify and calculate the landmarks by fast marching method and parabolic curve fitting. The accuracy and repeatability in CACVL group were compared with those in two manual landmarking groups [orthodontic experts (OE) group and orthodontic novices (ON) group]. The results showed that, as for the accuracy, there was no significant difference between CACVL group and OE group no matter in x-axis or y-axis (P>0.05), but there was significant difference between CACVL group and ON group, as well as OE group and ON group in both axes (P<0.05). As for the repeatability, CACVL group was more reliable than OE group and ON group in both axes. It is concluded that CACVL has the same or higher accuracy, better repeatability and less workload than manual landmarking methods. It's reliable for cervical parameters identification on the lateral cephalogram and cervical vertebral maturation prediction in orthodontic practice and research.

Objective To sequence and analyze the envelope (E) gene of type Ⅰ dengue virus isolated from Guangzhou in 2009 for tracing the infection source. Methods The serum samples were collected from patients diagnosed with dengue fever in Guangzhou area during 2009. Dengue virus was isolated and cultured in C6/36 cells.The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and then sequenced. The phylogenetic tree was drawn by neighbor-joining method. The bioinformatics analysis was performed by combining the phylogenetic information and the epidemiology data. Results Four strains of type Ⅰ dengue virus were isolated from 19 samples. E gene of these strains was amplified and sequenced. The phylogenetic analysis showed that 09/GZ/9104 strain and 09/GZ/9236 strain had identical nucleotide sequence and fell within the American/African group, 09/GZ/11534 stain and 09/GZ/11562 strain had similar sequence homology and fell within the Asian group. Conclusion The typeⅠdengue viruses in Guangzhou area in 2009 are imported, which belong to two genotypes and may come from two independent origins respectively.

AIM:To investigate the influence of continuous subculturing of human umbilical cord mesenchymal stem cells (hUC-MSCs) on the mRNA expression of all 23 family members of NOD-like receptors (NLRs), and to search for the way of improving the subculture quality of hUC-MSCs and increasing the quantity and safety in the experimental and clinical application .METHODS:Neonatal umbilical cord was collected to isolate and purify the hUC-MSCs with the colla-genase II digestion and adherence screening methods .These cells were continuously subcultured .The hUC-MSCs at pas-sage 3 and passage 28 were identified by flow cytometry and induced differentiation .The mRNA expression of NLRs in the passage 3 and passage 28 hUC-MSCs was detected by RT-qPCR.RESULTS: The cell phenotypes of both passage 3 and passage 28 hUC-MSCs were CD29 +/CD44 +/CD105 +/CD31 -/CD34 -/CD40 -/CD45 -/CD106 -/HLA-DR-, and both of the cells were induced into osteoblasts and adipocytes , which were conformed to the criteria of International Society for Cellular Therapy to define MSCs .All the NLR family members were expressed in passage 3 hUC-MSCs.NOD1, NLRC4, NLRC5, NLRP1, NLRP3, NLRP10, NAIP, NLRX1 and APAF1 at mRNA levels were highly expressed , and the rest were lowly expressed.When hUC-MSCs were subcultured to passage 28, NLRP10 mRNA was increased, NLRC5 mRNA and NLRX1 mRNA were hardly changed , and all of the rest members were decreased .The difference of NLRP1 mRNA expres-sion between passage 3 and passage 28 hUC-MSCs was observed with statistical significance (P<0.05).CONCLU-SION:The effects of subculturing on the expression of NLR family in hUC-MSCs are pleiotropic .It requires further investi-gation to confirm whether these effects are related to the proliferation , differentiation and immunomodulation of MSCs .

Objective To investigate the changes of the immunological factors in subcutaneous exudate and blood components of the rats receiving cutaneous scraping method,and to compare the changes of skin histopathological features before and after cutaneous scraping under microscope.Methods SD rats were randomly divided into two groups,cutaneous-scraping group and non-cutaneous-scraping group.And then each group was divided into three subgroups.The observation indexes included the levels of interleukin (IL)-1β,IL-6 and interferon gamma (IFN-γ) in the blood and the skin,routine blood examination,and skin histopathological features.Results In cutaneous-scraping group,the number of white blood cells in the blood and the levels of IL-1 β and IFN-γ in skin tissues were increased (P ＜ 0.05),the hemolysis rate was increased (P ＜ 0.05).However,the levels of IL-1β,IL-6 and IFN-γin the blood showed no obvious changes.Under the microscope,severe skin edema,vascular congestion and dilatation,and infiltration of inflammatory cells were found in the skin after cutaneous scraping.Conclusion The cutaneous scraping method can activate the immune response rapidly,and the immunological components of the subcutaneous exudate after cutaneous scraping are helpful to the disease treatment.

AIM:Toinvestigatetheeffectofnicotinicacidamide(NAA)ontheinfusiondamageofhuman umbilical cord mesenchymal stem cells ( hUC-MSCs) under the condition of instant blood-mediated inflammatory reaction ( IBMIR) .METHODS:Normal peripheral blood without anticoagulant at volume of 2.7 mL was mixed with 0.3 mL phys-iological saline (as blank group), CFSE labeled hUC-MSCs (1 ×106 cells in 0.3 mL as MSC group) and CFSE labeled hUC-MSCs (1 ×106 cells in 0.3 mL) preprocessed with NAA at concentration of 10 mmol/L for 24 h ( as MSC+NAA group) , respectively.The mixture was immediately injected into the improved Chandler Loop model, placed in 37℃water bath, and then started the peristaltic pump at the speed of 20 mL/min for 1 h.The number of CFSE labeled hUC-MSCs, platelets, white blood cells were counted and the concentration of complement C3a was measured before and after cycling, respectively.RESULTS: After 1 h circulation, the platelet dissipation rate were ( 29.96 ±10.88 )% in blank group, (77.76 ±19.29)% in MSC group all and (50.13 ±18.10)% in MSC +NAA group; and the leukocyte counts were (37.82 ±13.81)%in blank group, (64.57 ±17.08)% in MSC group and (41.52 ±17.26)% in MSC+NAA group. Compared with blank group, the differences of the dissipation rates in MSC group and MSC+NAA group all had statistical significance.The hUC-MSCs relative survival rate in MSC+NAA group was higher than that in MSC group.C3a concentra-tions in blank group, MSC group and MSC+NAA group were (206.27 ±58.10), (230.47 ±39.61) and (208.37 ± 40.66) μg/L, respectively.CONCLUSION:Co-circulating the mixture of hUC-MSCs with normal peripheral blood with-out anticoagulant in the improved Chandler Loop for 1 h depletes a large number of hUC-MSCs and blood components, and increases C3a, suggesting that this model can induce IBMIR.NAA has a protective effect on the hUC-MSCs in the infusion damage by inhibiting IBMIR, reducing the wastage of the blood components and enhancing the survival rate of the hUC-MSCs.

Objective:To investigate the learning curve of laparoscopic transanal total mesorectal excision (taTME) for rectal cancer operated by one or two surgery teams.Methods:The retrospective cross-sectional study was conducted. Based on the concept of real-world research, the clinical data of 1 458 patients undergoing laparoscopic rectal cancer taTME from 44 medical centers who were registered in the Chinese taTME registry collaborative (CTRC) database from May 2010 to May 2020 were collected. The 1 458 patients were divided into cohorts with one surgery team or two surgery teams according to the operation method. Patients with one surgery team underwent taTME by transabdominal operation and then by transanal operation. Patients with two surgery teams underwent taTME by transabdominal and transanal operation simultaneously with duration of the simutaneous operation time ≥30 minutes. The entire surgical process of patients with two surgery teams is not required to be performed by two surgery teams simutaneously. The clinical data were collected from the medical centers with similar operation amount according to the operation time sequence to analyze the difference between different operation stages and explore the learning curve. The operation time was taken as the parameter to carry out cumulative sum analysis and draw the learning curve of laparoscopic rectal cancer taTME in each medical center. The clinicopathological characteristics of patients from two medical centers with the largest difference in learning curves were analyzed. Observation indicators: (1) screening results of clinical data; (2) clinical data collection of patients with one surgery team; (3) surgical situations of laparoscopic rectal cancer taTME from the one surgery team in different operation stages; (4) learning curve of the one surgery team; (5) clinical data collection of patients with two surgery teams; (6) surgical situations of laparoscopic rectal cancer taTME from the two surgery teams; (7) learning curve of the two surgery teams. The cumulative sum was calculated by the CUSUM=∑i=1nXi-U, where Xi represented the operation time of each taTME, U represented the average operation time of all cases, and n represented the operation number. Fitting process was conducted on scatter plot of learning curves. Taking the apex of learning curve as the boundary, the learning curve was divided into two stages. The abscissa corresponding to the apex of learning curve was the number of operations that needed to be performed to cross the learning curve. Measurement data with normal distribution were represented as Mean±SD. Comparison between two groups was conducted using the t test and comparison between multiple groups was conducted using the ANOVA. Measurement data with skewed distribution were represented as M( P25,P75), and comparison between groups was conducted using the Mann-Whitney U test. Comparison of ordinal data was analyzed using the rank sum test. Count data were analyzed using the chi-square test or Fisher exact probability. Results:(1) Screening results of clinical data:the clinical data of 661 patients from 7 medical centers with one surgery team and two surgery teams were collected. (2) Clinical data collection of patients with one surgery team: the clinical data of 312 patients undergoing laparoscopic rectal cancer taTME from 5 medical centers were collected including 42 cases in the number 2 medical center, 97 cases in the number 20 medical center, 82 cases in the number 33 medical center, 35 cases in the number 37 medical center and 56 cases in the number 39 medical center, respectively. (3) Surgical situations of laparoscopic rectal cancer taTME from the one surgery team in different operation stages: three medical centers including the number 2, number 37 and number 39 medical center with close operation volume provided the clinical data of cases distributed in five operation stages. Among the five operation stages, the proportion of high-quality operation of total mesorectal excision (TME) was ≥17/18, the incidence of postoperative complications was ≤13.3%(4/30) and the incidence of anastomotic leakage was ≤10.0%(3/30). There was no significant difference in the TME quality, postoperative complications or anastomotic leakage among the five operation stages ( P>0.05). There was no significant difference in the operation time among the five operation stages ( χ2=6.950, P>0.05). (4) Learning curve of the one surgery team: the number of operations corresponding to the turning point of learning curve in number 2 and number 20 medical center was 22 and 39, respectively. The number of operations corresponding to the turning points of learning curve in number 33 and number 37 medical center was 15, 66 and 10, 28, respectively. The number of operations corresponding to the turning point of learning curve in number 39 medical center was 20. The overall curve of number 20 medical center was in line with the trend of learning curve and 39 cases of operations was the minimum number needed to cross the learning curve. The biggest difference in learning curve was shown between the number 20 and number 33 medical center. Cases with the gender of male or female, age, body mass index, cases classified as stage 1, stage 2, stage 3 or stage 4 of the American Society of Anesthesiologists (ASA) Classification, cases with neoadjuvant therapy, duration of postoperative hospital stay of the number 20 medical center were 77, 20, (60±10)years, 24 kg/m 2(22 kg/m 2, 26 kg/m 2), 1, 88, 8, 0, 8, 8, 11 days (9 days, 13 days), respectively, versus 51, 31, (64±11)years, 23 kg/m 2(21 kg/m 2, 26 kg/m 2), 0, 35, 43, 1, 31, 16 days (13 day, 21 day) of number 33 medical center, showing significant differences in the above indicators between the two medical centers ( χ2 =6.442, t=-2.265, Z=-2.032, -6.870, χ2 =22.120, Z=-8.408, P<0.05). (5) Clinical data collection of the two surgery teams: the clinical data of 259 patients undergoing laparoscopic rectal cancer taTME from 5 medical centers were collected, including 46 cases in the number 2 medical center, 47 cases in the number 8 medical center, 78 cases in the number 18 medical center, 43 cases in the number 33 medical center and 45 cases in the number 44 medical center, respectively. (6) Surgical situations of laparoscopic rectal cancer taTME from the two surgery teams: four medical centers including the number 2, number 8, number 33 and number 44 medical center with close operation volume provided the clinical data of cases distributed in four operation stages. Among the four operation stages, the proportion of high-quality operation of TME was ≥50.0%(13/26), the incidence of postoperative complications was ≤35.0%(14/40) and the incidence of anastomotic leakage was ≤22.5%(9/40). There was no significant difference in the TME quality, postoperative complications or operation time among the four operation stages ( χ2 =3.252, 4.733, 8.848, P>0.05). There was a significant difference in the incidence of anastomotic leakage among the four operation stages ( P<0.05). (7) Learning curve of the two surgery teams: the number of operations corresponding to the turning point of learning curve in number 2 and number 8 medical center was 28 and 16, respectively. The number of operations corresponding to the turning points of learning curve in number 18, number 33 and number 44 medical center was 12 and 58, 10 and 36, 14 and 36, respectively. The overall curve of number 2 medical center was in line with the trend of learning curve and 28 cases of operations was the minimum number needed to cross the learning curve. The biggest difference in learning curve was shown between the number 2 and number 33 medical center. The age and cases with tumor in stage T0 and (or) Tis, stage T1, stage T2, stage T3 or stage T4 of the T staging of the number 2 and number 33 medical center were (60±12)years, 3, 1, 9, 11, 20 and (65±10)years, 2, 3, 22, 15, 0, respectively, showing significant differences in the above indicators between the two medical centers ( t=-2.280, Z=-4.033, P<0.05). Conclusion:Thirty-nine cases of operations was the minimum number for the one surgery team to cross the learning curve of laparoscopic rectal cancer taTME and 28 cases of operations was the minimum number for the two surgery teams to cross the learning curve of laparoscopic rectal cancer taTME.

The accuracy and repeatability of computer aided cervical vertebra landmarking (CACVL) were investigated in cephalogram.120 adolescents (60 boys,60 girls) aged from 9.1 to 17.2 years old were randomly selected.Twenty-seven landmarks from the second to fifth cervical vertebrae on the lateral cephalogram.were identified.In this study,the system of CACVL was developed and used to identify and calculate the landmarks by fast marching method and parabolic curve fitting.The accuracy and repeatability in CACVL group were compared with those in two manual landmarking groups [orthodontic experts (OE) group and orthodontic novices (ON) group].The results showed that,as for the accu racy,there was no significant difference between CACVL group and OE group no matter in x-axis or y-axis (P＞0.05),but there was significant difference between CACVL group and ON group,as well as OE group and ON group in both axes (P＜0.05).As for the repeatability,CACVL group was more reliable than OE group and ON group in both axes.It is concluded that CACVL has the same or higher accuracy,better repeatability and less workload than manual landmarking methods.It's reliable for cervical parameters identification on the lateral cephalogram and cervical vertebral maturation prediction in orthodontic practice and research.

Objectives: The purpose of this study was to compare differences in facial soft tissue thickness in three-dimensional (3D) images before and after orthognathic surgery in patients with skeletal Class III malocclusion and to obtain a better understanding of the relationship between hard and soft tissue changes after surgery.Materials and method: The present retrospective study included 31 patients with skeletal Class III malocclusion with mandibular chin deviation greater than 4 mm who had undergone cone-beam computed tomography before and 6 months after surgery. Seven bilateral points were established. Measurements were taken from software-generated multiplanar reconstructions. The predictor variables were timing (pre- and postoperatively) and side (deviated vs.nondedicated). A regression model and correlation analysis were conducted for statistical analysis. Results: The difference of bilateral facial soft tissue thickness was statistically significantly different between deviated and nondeviated sides (P < 0.05), with lower values observed on the deviated side. The soft tissue thickness has become nearly symmetric at local regions of the lower thirds of the face after orthognathic surgery. However, most measurements showed a negative correlation between changes in soft tissue thickness and changes in bone tissues. Conclusions Skeletal Class III malocclusion with facial asymmetry is accompanied by differences in soft tissue thickness when comparing Dev and N-Dev sides of the posterior region of the mandible, where soft tissues are thinner on the Dev side. Soft tissue thickness can compensate for or camouflage the underlying asymmetric mandible. In addition, the asymmetric soft tissue thickness on the lower third of the face can be partially improved by orthognathic surgery, but the amount of soft tissue thickness change is not consistent with that of hard tissue positional change.

Objective:To investigate the incidence, clinical characteristics and risk factors for hip fractures in patients within two years after stroke onset.Methods:A total of 332 persons with first-onset stroke from the neurology department of our hospital between 1 June 2013 and 31 December 2014 were recruited and were divided into the hip fracture group and the non-hip fracture group.Clinical characteristics were recorded.Vision was tested as normal or impaired.Patients were accessed by the National Institutes of Health Stroke Scale(NIHSS), Behavioral Inattention Test, Baking Tray Task, Mini-Mental State Examination(MMSE), Birgitta Lindmark(BL)motor assessment scale, Berg Balance Scale(BBS), Timed Up & Go(TUG)Scale, and Stops Walking When Talking(SWWT)Scale.The clinic characteristics and risk factors for hip fractures were compared between the two groups after a 2-year follow-up.The accuracy of risk factors for fracture prediction was assessed by the sensitivity, specificity, and positive and negative predictive values.Results:Of 332 patients with stroke, 16 cases fractured their hips within two years after stroke onset, which corresponded to an incidence of 33‰/year(95% CI: 15‰/year-50‰/year). The 2-year mortality rate was 44%(95% CI: 25%-60%)and 48%(95% CI: 42%-54%)in patients with and without hip fractures respectively( χ2=0.036, P=0.724). The mean survival time for patients with and without hip fracture was 2.72 years(95% CI: 1.45-2.79)and 2.21 years(95% CI: 1.48-2.34)respectively.The proportions of patients with previous fractures history( χ2=16.780, P=0.041)and impaired vision( χ2=11.210, P=0.027), MMSE scale score( U=14.220, P=0.031), TUG ≥ 15 s( χ2=18.560, P=0.000)were higher, and SWWT( χ2=20.340, P=0.000)was lower in the hip fracture group than in the non-hip fracture group.The negative predictive values of previous fractures history, impaired vision, TUG and SWWT were higher than their positive predictive value.The specificities of previous fractures history, impaired vision, and SWWT were higher than their sensitivities.And the sensitivity of TUG was higher than its specificity. Conclusions:Hip fractures after stroke are common in elderly patients.Fractures often occur during daytime at home in daily activities.The previous fractures history, visual and cognitive dysfunction and impaired functional mobility are risk factors for hip fractures.We should take measures to prevent falls according to the relevant factors.Among the test scales, the timed up & go(TUG)scale could much more accurately identify patients at high risk for hip fractures.