[Effects of gelatin methacrylate anhydride hydrogel loaded with small extracellular vesicles derived from human umbilical cord mesenchymal stem cells in the treatment of full-thickness skin defect wounds in mice].

Objective: To investigate the effects of gelatin methacrylate anhydride (GelMA) hydrogel loaded with small extracellular vesicles derived from human umbilical cord mesenchymal stem cells (hUCMSCs-sEVs) in the treatment of full-thickness skin defect wounds in mice. Methods: This study was an experimental study. hUCMSCs-sEVs were extracted by ultracentrifugation, their morphology was observed through transmission electron microscope, and the expression of CD9, CD63, tumor susceptibility gene 101 (TSG101), and calnexin was detected by Western blotting. The human umbilical vein endothelial cells (HUVECs), the 3rd and 4th passages of human epidermal keratinocytes (HEKs) and human dermal fibroblasts (HDFs) were all divided into blank control group (routinely cultured) and hUCMSC-sEV group (cultured with the cell supernatant containing hUCMSCs-sEVs). The cell scratch test was performed and the cell migration rates at 6, 12, and 24 h after scratching were calculated, the cell Transwell assay was performed and the number of migration cells at 12 h after culture was calculated, and the proportion of proliferating cells was detected by 5-acetylidene-2'-deoxyuridine and Hoechst staining at 24 h after culture, with sample numbers being all 3. The simple GelMA hydrogel and the GelMA hydrogel loaded with hUCMSCs-sEVs (hereinafter referred to as hUCMSC-sEV/GelMA hydrogel) were prepared. Then the micromorphology of 2 kinds of hydrogels was observed under scanning electron microscope, the distribution of hUCMSCs-sEVs was observed by laser scanning confocal microscope, and the cumulative release rates of hUCMSCs-sEVs at 0 (immediately), 2, 4, 6, 8, 10, and 12 d after soaking hUCMSC-sEV/GelMA hydrogel in phosphate buffer solution (PBS) were measured and calculated by protein colorimetric quantification (n=3). Twenty-four 6-week-old male C57BL/6J mice were divided into PBS group, hUCMSC-sEV alone group, GelMA hydrogel alone group, and hUCMSC-sEV/GelMA hydrogel group according to the random number table, with 6 mice in each group, and after the full-thickness skin defect wounds on the back of mice in each group were produced, the wounds were performed with PBS injection, hUCMSC-sEV suspenson injection, simple GelMA coverage, and hUCMSC-sEV/GelMA hydrogel coverage, respectively. Wound healing was observed on post injury day (PID) 0 (immediately), 4, 8, and 12, and the wound healing rates on PID 4, 8, and 12 were calculated, and the wound tissue was collected on PID 12 for hematoxylin-eosin staining to observe the structure of new tissue, with sample numbers being both 6. Results: The extracted hUCMSCs-sEVs showed a cup-shaped structure and expressed CD9, CD63, and TSG101, but barely expressed calnexin. At 6, 12, and 24 h after scratching, the migration rates of HEKs (with t values of 25.94, 20.98, and 20.04, respectively), HDFs (with t values of 3.18, 5.68, and 4.28, respectively), and HUVECs (with t values of 4.32, 19.33, and 4.00, respectively) in hUCMSC-sEV group were significantly higher than those in blank control group (P<0.05). At 12 h after culture, the numbers of migrated HEKs, HDFs, and HUVECs in hUCMSC-sEV group were 550±23, 235±9, and 856±35, respectively, which were significantly higher than 188±14, 97±6, and 370±32 in blank control group (with t values of 22.95, 23.13, and 17.84, respectively, P<0.05). At 24 h after culture, the proportions of proliferating cells of HEKs, HDFs, and HUVECs in hUCMSC-sEV group were significantly higher than those in blank control group (with t values of 22.00, 13.82, and 32.32, respectively, P<0.05). The inside of simple GelMA hydrogel showed a loose and porous sponge-like structure, and hUCMSCs-sEVs was not observed in it. The hUCMSC-sEV/GelMA hydrogel had the same sponge-like structure, and hUCMSCs-sEVs were uniformly distributed in clumps. The cumulative release rate curve of hUCMSCs-sEVs from hUCMSC-sEV/GelMA hydrogel tended to plateau at 2 d after soaking, and the cumulative release rate of hUCMSCs-sEVs was (59.2±1.8)% at 12 d after soaking. From PID 0 to 12, the wound areas of mice in the 4 groups gradually decreased. On PID 4, 8, and 12, the wound healing rates of mice in hUCMSC-sEV/GelMA hydrogel group were significantly higher than those in the other 3 groups (P<0.05); the wound healing rates of mice in GelMA hydrogel alone group and hUCMSC-sEV alone group were significantly higher than those in PBS group (P<0.05). On PID 8 and 12, the wound healing rates of mice in hUCMSC-sEV alone group were significantly higher than those in GelMA hydrogel alone group (P<0.05). On PID 12, the wounds of mice in hUCMSC-sEV/GelMA hydrogel group showed the best wound epithelization, loose and orderly arrangement of dermal collagen, and the least number of inflammatory cells, while the dense arrangement of dermal collagen and varying degrees of inflammatory cell infiltration were observed in the wounds of mice in the other 3 groups. Conclusions: hUCMSCs-sEVs can promote the migration and proliferation of HEKs, HDFs, and HUVECs which are related to skin wound healing, and slowly release in GelMA hydrogel. The hUCMSC-sEV/GelMA hydrogel as a wound dressing can significantly improve the healing speed of full-thickness skin defect wounds in mice.

[Reassessment of practice of Chinese surgeons since introduction of the watch and wait strategy after neoadjuvant therapy for rectal cancer].

Objective: To investigate perspectives and changes in treatment selection by Chinese surgeons since introduction of the watch-and-wait approach after neoadjuvant therapy for rectal cancer. Methods: A cross-sectional survey was conducted using a questionnaire distributed through the "Wenjuanxing" online survey platform. The survey focused on the recognition and practices of Chinese surgeons regarding the strategy of watch-and-wait after neoadjuvant therapy for rectal cancer and was disseminated within the China Watch-and-Wait Database (CWWD) WeChat group. This group targets surgeons of deputy chief physician level and above in surgical, radiotherapy, or internal medicine departments of nationally accredited tumor-specialist or comprehensive hospitals (at provincial or municipal levels) who are involved in colorectal cancer diagnosis and treatment. From 13 to 16 December 2023, 321 questionnaires were sent with questionnaire links in the CWWD WeChat group. The questionnaires comprised 32 questions encompassing: (1) basic physician characteristics (including surgical volume); (2) assessment methods and criteria for clinical complete response (cCR); (3) patients eligible for watch-and-wait; (4) neoadjuvant therapies and other measures for achieving cCR; (5) willingness to implement watch-and-wait and factors influencing that willingness; (6) risks and monitoring of watch-and-wait; (7) subsequent treatment and follow-up post watch-and-wait; (8) suggestions for development of the CWWD. Descriptive statistics were employed for data analysis, with intergroup comparisons conducted using the χ2 or Fisher's exact probability tests. Results: The response rate was 31.5%, comprising 101 responses from the 321 individuals in the WeChat group. Respondents comprised 101 physicians from 70 centers across 23 provinces, municipalities, and autonomous regions nationwide, 85.1% (86/101) of whom represented provincial tertiary hospitals. Among the respondents, 87.1% (88/101) had implemented the watch-and-wait strategy. The approval rate (65.6%, 21/32) and proportion of patients often informed (68.8%, 22/32) were both significantly higher for doctors in oncology hospitals than for those in general hospitals (27.7%, 18/65; 32.4%, 22/68) (χ2=12.83, P<0.001; χ2=11.70, P=0.001, respectively). The most used methods for diagnosing cCR were digital rectal examination (90.1%, 91/101), colonoscopy (91.1%, 92/101), and rectal T2-weighted magnetic resonance imaging (86.1%, 87/101). Criteria used to identify cCR comprised absence of a palpable mass on digital rectal examination (87.1%, 88/101), flat white scars or new capillaries on colonoscopy (77.2%, 78/101), absence of evident tumor signals on rectal T2-weighted sequences or T2WI low signals or signals equivalent to the intestinal wall (83.2%, 84/101), and absence of tumor hyperintensity on diffusion-weighted imaging with no corresponding hypointensity on apparent diffusion coefficient maps (66.3%, 67/101). As for selection of neoadjuvant regimen and assessment of cCR, 57.4% (58/101) of physicians preferred a long course of radiotherapy with or without induction and/or consolidation capecitabine + oxaliplatin, whereas 25.7% (26/101) preferred immunotherapy in combination with chemotherapy and concurrent radiotherapy. Most (96.0%, 97/101) physicians believed that the primary lesion should be assessed ≤12 weeks after completion of radiotherapy. Patients were frequently informed about the possibility of achieving cCR after neoadjuvant therapy and the strategy of watch-and-wait by 43.6% (44/101) of the responding physicians and 38.6% (39/101) preferred watch-and-wait for patients who achieved cCR or near cCR after neoadjuvant therapy for rectal cancer. Capability for multiple follow-up evaluations (70.3%, 71/101) was a crucial factor influencing physicians' choice of watch-and-wait after cCR. The proportion who patients who did not achieve cCR and underwent surgical treatment was lower in provincial tertiary hospitals (74.2%, 23/31) than in provincial general hospitals (94.5%, 52/55) and municipal hospitals (12/15); these differences are statistically significant (χ2=7.43, P=0.020). The difference between local recurrence and local regrowth was understood by 88.1% (89/101) of respondents and 87.2% (88/101) agreed with monitoring every 3 months for 5 years. An increase in local excision or puncture rates to reduce organ resections in patients with pCR was proposed by 64.4% (65/101) of respondents. Conclusion: Compared with the results of a previous survey, Chinese surgeons' awareness of the watch-and-wait concept has improved significantly. Oncologists in oncology hospitals are more aware of the concept of watch-and-wait.

Removal of intraglandular calculi in Wharton's duct: clinical outcome and treatment algorithm.

The aim of this study was to propose a treatment strategy for intraglandular submandibular calculi based on calculus site. Seventy-three consecutive patients with impalpable intraglandular submandibular calculi were enrolled retrospectively. The calculi were classified as either post-hilar type, central type, or superficial type. Treatment approaches included transoral duct slitting (TDS), interventional basket retrieval (IBR), intraductal laser lithotripsy (ILL), and transcervical lithotomy (TCL). Complete calculus removal with gland preservation was achieved in 64 patients (87.7%). The success rate for post-hilar, central, and superficial calculi was 86.4% (51/59), 90.9% (10/11), and 100% (3/3), respectively. The treatment approach applied in patients with treatment success was TDS in 32 cases, IBR in 20, ILL in nine, and TCL in three. During follow-up (median 17.3 months), one patient experienced gland atrophy and three had ductal stenosis; the remaining 60 patients (93.8%, 60/64) had good clinical outcomes. In the eight failure cases operated by TDS, the deeply situated calculi could not be detached despite the parenchymal incision in five cases, while the procedure was ceased due to the patient's inability to cooperate in the other three cases. In the remaining failure case, the submandibular gland was sacrificed after calculus extraction via TCL. Application of the proposed treatment algorithm might help preserve gland function in patients with intraglandular submandibular calculi.

Comparison of two transcutaneous approaches for the removal of impacted parotid stones.

The aim of this study was to comparatively evaluate the indications and treatment outcomes of two transcutaneous approaches for the removal of impacted parotid stones. Sixty-eight consecutive patients with impacted parotid stones underwent endoscopy-assisted lithotomy via a direct mini-incision or a peri-auricular flap. Clinical safety and outcomes were evaluated. Complete stone extraction was achieved in all patients. In the mini-incision group (52 patients), the stones were in the middle third of the main duct in 31 patients, at the hilum in 16, and in the intraglandular duct in five. In the flap group (16 patients), they were in the middle third of the main duct in one patient, at the hilum in seven, and in the intraglandular duct in eight. Salivary fistula occurred in five mini-incision group patients (9.6%) and four flap group patients (25%). The clinical outcome in the mini-incision group (47 patients, median 25 months of follow-up) was good in 28 patients, fair in 13, and poor in six (12.8%). The clinical outcome in the flap group (16 patients, median 84 months of follow-up) was good in nine patients, fair in five, and poor in two (12.5%). The direct mini-incision approach was found to be safe and effective for impacted stones in the middle third, hilum, and proximal third of the main duct, while the peri-auricular approach would be best reserved for deeper intraglandular stones.

[Summary of community smoking cessation intervention theory].

This paper analyzes the literatures about community-based smoking cessation interventions in recent decades and introduces the effectiveness of cessation interventions developed under different theoretical frameworks applied in the community. Because of the severe smoking prevalence in China and the shortage of existing smoking cessation services, the application of smoking cessation services in reducing the smoking rate in Chinese is discussed to provide a reference for the theoretical framework and practical application of community smoking cessation intervention research in China.

[Clinicopathological analysis of 105 patients with fibrous dysplasia of cranio-maxillofacial region].

OBJECTIVE: To compare the clinicopathologic features and prognosis of the different types of fibrous dysplasia (FD) of cranio-maxillofacial region, so as to provide a new reference for clinicians to treat these patients and make prognostic judgement. METHODS: Clinical records, radiographic data and pathological information of 105 patients diagnosed with FD or McCune-Albright syndrome (MAS) at the Department of Oral Pathology, Peking University Hospital of Stomatology from January 2013 to December 2020 were collected. The patients were divided into 4 groups: monostotic FDs, polyostotic FDs, MAS and a specific type called craniofacial fibrous dysplasia (CFD) limited in the craniofacial region. The clinicopathological characteristics, treatment and follow-up data of each type were analyzed. RESULTS: Of all the 105 patients, 46 were males and 59 were females, with a male-to-female ratio of 1 ∶1.3. The onset age ranged from 0 to 56 years and the median age was 12 years. On the basis of different involvement conditions, 4 types were divided. The most common type was monostotic FDs (43 cases, 40.95%), including maxilla (29 cases), mandibular (12 cases) and zygoma (2 cases). 32 cases (30.48%) were diagnosed with polyostotic FDs, 7 cases (6.67%) were MAS, and 23 cases (21.90%) were CFDs confirmed by computed tomography (CT) analysis. CFD was clearly distinct from other types of FD, such as the patient gender and the serum alkaline phosphatase level in peripheral blood before operative surgery. The pathologic findings of various types FD were quite similar, whilst the predominant fibrous tissue hyperplasia could be observed in polyostotic FDs and MAS types. CONCLUSION: The clinicopathologic features of FD in the cranio-maxillofacial region are different from the FD lesions in other parts of the body. The clinicopathological features of CFD are significantly different from those of monostotic and polyostotic FDs in the cranio-maxillofacial region. Therefore, the clinicians should pay attention to distinguish CFD in clinic, imaging and pathology aspects, so as to further clarify its features in clinic management and prognosis.

A bimodal burst energy distribution of a repeating fast radio burst source.

The event rate, energy distribution and time-domain behaviour of repeating fast radio bursts (FRBs) contain essential information regarding their physical nature and central engine, which are as yet unknown1,2. As the first precisely localized source, FRB 121102 (refs. 3-5) has been extensively observed and shows non-Poisson clustering of bursts over time and a power-law energy distribution6-8. However, the extent of the energy distribution towards the fainter end was not known. Here we report the detection of 1,652 independent bursts with a peak burst rate of 122 h-1, in 59.5 hours spanning 47 days. A peak in the isotropic equivalent energy distribution is found to be approximately 4.8 × 1037 erg at 1.25 GHz, below which the detection of bursts is suppressed. The burst energy distribution is bimodal, and well characterized by a combination of a log-normal function and a generalized Cauchy function. The large number of bursts in hour-long spans allows sensitive periodicity searches between 1 ms and 1,000 s. The non-detection of any periodicity or quasi-periodicity poses challenges for models involving a single rotating compact object. The high burst rate also implies that FRBs must be generated with a high radiative efficiency, disfavouring emission mechanisms with large energy requirements or contrived triggering conditions.

Comparison of hepatic epithelioid angiomyolipoma and non-hepatitis B, non-hepatitis C hepatocellular carcinoma on contrast-enhanced ultrasound.

PURPOSE: The purpose of this study was to retrospectively compare the imaging features of hepatic epithelioid angiomyolipoma (HEAML) to those of hepatocellular carcinoma negative for hepatitis B surface antigen and hepatitis C antibody (NBNC-HCC) on contrast-enhanced ultrasound (CEUS) with sulphur hexafluoride microbubbles. MATERIAL AND METHODS: Twenty-two patients (4 men, 18 women) with a mean age of 42.6±10.2 (SD) years (range: 22-63 years) with histopathologically confirmed HEMAL were included in the study. Forty-four patients (30 men, 14 women) with a mean age of 57.3±15.9 years (range: 19-85 years) with histopathologically confirmed NBNC-HCC were randomly selected from our institution's database as a control group. The CEUS characteristics of the two groups were compared. RESULTS: On conventional ultrasound, significant differences in tumor diameter were found between HEAML (4.0±2.0 [SD] cm; range: 1.3-8.9cm) and NBNC-HCC (8.4±4.4 [SD] cm; range: 1.6-18cm) (P<0.001) as well as in degrees of enhancement during the portal (P=0.001) and late phases (P=0.003), contrast distribution (P<0.001) and absence of pseudocaspule (P<0.001). On CEUS, hyperenhancement during the arterial phase was observed in 21/22 (95.5%) HEAMLs and in 43/44 (97.7%) NBNC-HCCs (P>0.999). Homogeneous enhancement was more frequent in HEAMLs (20/22; 90.9%) than in NBNC-HCCs (13/44; 29.6%) (P<0.001). Pseudocapsule was observed in 0/22 HEAMLs (0.0%) and in 36/44 NBNC-HCCs (81.8%) (P=0.017). A prolonged enhancement was observed in 5/22 HEAMLs (22.7%) and in 0/44 NBNC-HCCs (0.0%) (P<0.001) during the late phase. CONCLUSION: CEUS with sulphur hexafluoride microbubbles is helpful in discriminating between HEAML and NBNC-HCC. Homogeneous enhancement and lack of pseudocapsule are suggestive features for the diagnosis of HEAML.

[Clinicopathological analysis of 844 cases of odontogenic keratocysts].

OBJECTIVE: To investigate the clinicopathologic features and prognostic factors in odontogenic keratocyst (OKC), and to provide new reference for clinic treatment and management of these patients. METHODS: Clinicopathological data of 844 cases initially diagnosed as or associated with OKC at Department of Oral Pathology, Peking University Hospital of Stomatology from 2000 to 2018 were collected. The cases were divided into 4 groups: sporadic OKCs (intraosseous, cystic lesion irrelevant to nevoid basal cell carcinoma syndrome), syndromic OKCs, solid OKCs and peripheral OKCs. The patients were follow-up for 6 to 216 months and the factors that might relate to recurrence were analyzed. RESULTS: There were 805 cases (95.4%) of sporadic OKCs, 32 cases (3.8%) of syndromic OKCs, 3 cases of solid OKCs and 4 cases of peripheral OKCs. The main age of sporadic OKCs was 36.03 years with the peak at the second and third decades. Ratio of male and female was 1.27:1. The predilection site was the molar and ramus area of mandibular (56.2%). In the study, 428 cases (71.2%) were unilocular in radiography while 28.8% were multilocular. The recurrent rate of enucleation with the follow-up was 20.1% (118/588) while most of them occurred in 1-3 years after surgery. The recurrent rate of multilocular patients (39.0%) was significantly higher than that of the unilocular. Enucleation after marsupialization (43 cases) or enucleation only (545 cases) showed no difference in recurrence (P>0.05). The syndromic OKCs was younger (main 20.97) and preferred to be multiple compared with sporadic OKCs (30/32, 93.7%). The predilection site was also molar and ramus area of mandibular (41.7%). Age and gender distribution of multiple cases had no significant difference with those in sporadic OKCs. More daughter cysts and epithelial islands were seen (56.3% and 17.9%). Furthermore, the recurrent rate was significantly higher than that of the sporadic OKCs (13/29, 44.9%). But there was no evidence of recurrent-related factors. The age of solid and peripheral OKCs, averaged at 45.00 and 65.75 years, were older than others. Four of peripheral OKCs showed no recurrence after enucleation. CONCLUSION: The recurrence rate of sporadic OKCs after enucleation is 20.1%. The multilocular lesions prefer to be recurrent. There is no significant difference of recurrence with enucleation only or enucleation after marsupialization. Compared with sporadic OKCs, the syndromic patients are younger and easier to be multiple. It tends to be recurrent frequently and rapidly. There are no related factors about recurrence of syndromic patients. The clinicians should considerate comprehensively and make an individual management of therapy and follow-up. Solid and peripheral OKCs are rare and older.

[Parotid CT imaging reporting and data system: A preliminary study].

OBJECTIVE: To establish a Parotid Imaging Reporting and Data System (PI-RADS) for CT diagnosis of the parotid gland neoplasms and to investigate the clinical applicable value and feasibility of PI-RADS. METHODS: Patients who had been diagnosed with primary parotid gland neoplasms and had received surgical treatments in Peking University School and Hospital of Stomatology during the period of January 2013 to December 2016 were included in this study. The diagnoses were confirmed by the postoperative pathological examinations in all the patients. The CT imaging data of all patients were retrospectively reviewed and analyzed by two readers in consensus. Imaging characteristics related to the parotid neoplasms were extracted and quantified. Based on comprehensive analysis of the imaging characteristics, the probabilities of the benign and malignant neoplasms were evaluated and classified into six grades, PI-RADS 1-6 (PI-RADS 1: normal parotid gland; PI-RADS 2: confidently benign lesions; PI-RADS 3: probably benign lesions without confirmed evidence of malignancy; PI-RADS 4: suspected malignancy without sufficient evidence of malignancy; PI-RADS 5: confidently malignant lesions; PI-RADS 6: lesions with confirmed pathological evidence of malignancy). RESULTS: A total of 897 patients with 1 003 parotid lesions were included. The lesions included 905 benign and 98 malignant lesions. The proportions of the malignancies in PI-RADS 2, PI-RADS 3, PI-RADS 4 and PI-RADS 5 according to the two readers in consensus were 0.4%, 5.7%, 35.5% and 96.7% respectively. The overall Cohen's Kappa test showed medium consistency between the two independent researchers (κ=0.614, P<0.001, 95%CI: 0.569-0.695). Pearson Chi-square test showed that the proportions of malignancies increased with the diagnostic PI-RADS grades (Cochran-Armitage trend test, Z=-15.579, P<0.001). The results of Pearson Chi-square tests showed significant differences between the grades [PI-RADS 2 and 3 (χ²=12.048, P=0.001); PI-RADS 3 and 4 (χ²=75.231, P<0.001); PI-RADS 4 and 5 (χ²=32.266, P<0.001)]. CONCLUSION: PI-RADS can be used to evaluate the risk of malignancy and will be helpful to improve the imaging diagnosis and clinical treatment of parotid gland neoplasms.

[Application of computer-assisted design for anterolateral thigh flap in oral and maxillofacial reconstruction].

OBJECTIVE: To investigate the feasibility and accuracy of using digital technology to design anterolateral thigh flap (ALTF) in oral and maxillofacial defect reconstruction. METHODS: Ten cases underwent oral and maxillofacial defects reconstruction with ALTFs in Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology from June 2019 to Oct. 2019 were enrolled. There were 7 males and 3 females with the mean age of 47.1 years. Preoperative high frequency color Doppler ultrasound examination was performed to detect the perforators of ALTF. CT data of the thigh was imported in DICOM (digital imaging and communications in medicine) format to the Proplan CMF 3.0 software (Materalise, Belgium), then virtual harvest of ALTF was performed according to the points of perforators detected by high frequency color Doppler ultrasound and the virtual flap volume was calculated by Proplan CMF 3.0 software. ALTF was harvested followed by preoperative virtual design, and the actual flap volume of ALTF was measured by the draining method during the surgery. Finally, the accuracy rate of using high frequency color Doppler ultrasound to detect perforators of ALTFs was calculated, and the differences between the virtual flap volume measured by Propaln CMF 3.0 software and the actual volume of ALTF by the draining method were compared using paired samples T test. RESULTS: Fifteen perforators in the flaps area of 10 patients who underwent oral and maxillofacial defects reconstruction with ALTFs were detected by high frequency color Doppler ultrasound, and 16 perforators were identified during the surgery, with the accuracy rate of 87.5%. The flaps size ranged from 5 cm×7 cm to 8 cm×15 cm, all the 10 flaps survived. The donor sites were primarily closed without skin graft, and no surgery complication was found on the donor site. The mean flap volume measured by Propaln CMF 3.0 software was 71.4 cm³ (range: 36.1-188.4 cm³), and the mean volume measured by the draining method was 70.7 cm³ (range: 38.3-172.5 cm³). There was no significant difference between the virtual flap volume measured by Propaln CMF 3.0 software and the actual volume measured by draining method (t=0.318; P=0.758). CONCLUSION: Preoperative virtual design of ALTF has good feasibility and accuracy and can be used to guide the harvest of ALTF during operation.

MicroRNA-101-5p inhibits the growth and metastasis of cervical cancer cell by inhibiting CXCL6.

OBJECTIVE: The objective of this study is to explore the biological roles of microRNA-101-5p (miR-101-5p) in the growth and metastasis of cervical cancer. PATIENTS AND METHODS: The levels of miR-101-5p and chemokine (C-X-C motif) ligand 6 (CXCL6) in cervical cancer tissues and cells were detected using the quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay. The proliferation, colony formation, migration, and invasion assays were conducted using miR-101-5p transfected cervical cancer cell. The expression of CXCL6 was measured by the immunoblotting assay. Xenograft model was constructed to reveal the precise roles of miR-101-5p in the growth of cervical cancer cell in vivo. RESULTS: MiR-101-5p was down-regulated in cervical cancer tissues when compared to the normal controls. The levels of miR-101-5p were higher in cervical cancer cells (SiHa, Caski, C-4-I, C-33 A) than that in the human cervical surface epithelial cell line, HcerEpic. Over-regulation of miR-101-5p inhibited the aggressiveness phenotypes of a cervical cancer cell in vitro. Furthermore, over-regulation of miR-101-5p reduced the tumor growth of cervical cancer cell in vivo. CXCL6 was the target protein of miR-101-5p in cervical cancer as demonstrated by luciferase reporter assay. The mRNA level of CXCL6 was negatively associated with the miR-101-5p level in cervical cancer tissue. Finally, the rescue experiments suggested that the inhibitory role of miR-101-5p was mediated by regulating the expression of CXCL6 in cervical cancer. CONCLUSIONS: These findings indicated that the over-regulation of miR-101-5p suppressed the progression of cervical cancer by targeting CXCL6 and might function as a potential therapeutic target for cervical cancer.

[Endoscopy-assisted transoral removal of deep hilar and intraparenchymal stones in the Wharton's duct].

Objective: To investigate the different surgical approaches and long-term outcomes of endoscopy-assisted transoral removal of deep hilar and intraparenchymal stones in the Wharton's duct. Methods: From January 2008 to March 2018, 481 consecutive patients with deep hilar and intraparenchymal calculi in the Wharton's duct underwent endoscopy-assisted transoral removal at Deparment of Oral and Maxillofacial Radiology, Peking University School and Hospital of Stomatology. There were 250 males and 231 females. Their ages ranged from 9-86 years. We operated 476 patients under local anesthesia on an outpatient basis, and the remaining 5 were operated under general anesthesia. On the basis of ultrasonography, spiral CT, sialography and endoscopy, the calculi were classified into 4 types: hilum stones (located at the hilum or proximally with a distance <5 mm from the hilum), infra-hilum stones (intra-glandular stones with a distance of 5-10 mm from the hilum), intraparenchymal stones (with a distance ≥10 mm from the hilum), and multiple stones (concomitant hilum and intra-glandular stones). The treatment approaches included: hilum duct slitting, intraparenchymal duct slitting, submandibulotomy and intraductal retrieval. The success rate, immediate safety and effectiveness of different types of stones were evaluated. After surgery, the patients were followed up, and gland function was analyzed on the basis of clinical symptoms and signs. Results: The calculi sizes varied from 3 to 25 mm, with a mean of 7.8 mm. The calculi were located in the right submandibular gland in 259 patients, in the left submandibular gland in 219 patients and in bilateral glands in 3 patients. The calculi were successfully removed in 446 glands, with a success rate of 92.1% (446/484). The success rate varied according to the stone sites: 97.8% (363/371) for hilum stones, 64.4% (29/45) for infra-hilum stones, 4/16 for intraparenchymal stones and 96.2% (50/52) for multiple stones. The main treatment methods applied included hilum duct slitting in 347 glands, intraparenchymal duct slitting in 13, submandibulotomy in 4, intraductal retrieval in 73, and hilum duct slitting accompanied by intraductal retrieval in 9. Ductal breakage occurred in 2 glands. All patients complained of mild to moderate pain with a duration of 3-7 days. Nine had temporal lingual nerve injury. During 3-120 months' follow-up (mean 36 months) of the total 484 glands, 1.6% (7/446) developed ranula, 1.3% (6/446) experienced obturation of the main duct and 2.0% (9/446) had recurrent stones. The remaining 95.1% (424/446) glands were symptom-free with good function. Conclusions: Endoscopy-assisted transoral removal of deep hilar and intraparenchymal submandibular calculi is a safe and effective gland-preserving procedure. According to the depth, size and number of the calculi, variant surgical approaches should be attempted to maximize the success rate and to minimize the side effects.

[Expression and potential clinical significance of cytidine triphosphate: phosphocholine cytidylyltransferase-α in oral squamous cell carcinoma].

Objective: To examine the expression and potential clinical significance of CCT (cytidine triphosphate: phosphocholine cytidylyltransferase)-α in oral squamous cell carcinoma (OSCC). Methods: Fifty-eight OSCC and paired adjacent non-malignant epithelia samples (between May 2016 and July 2016) were obtained from dental center, Second Xiangya Hospital, Central South University. CCT-α expression was examined by immunohistochemistry. The relationship between CCT-α and clinicopathological features of OSCC patients was analyzed. Quantitative real-time PCR and Western blot were performed to measure the expression of CCT-α mRNA and protein level in several OSCC cell line and two normal oral epithelial cell line. Results: Immunohistochemistry showed that CCT-α positive staining was found in cell nuclear of OSCC cells and adjacent epithelial cells. CCT-α was positively expressed in OSCC, which was significantly higher than that adjacent to carcinoma tissues (P=0.000). The expression of CCT-α in oral squamous cell carcinoma was correlated with smoking, alcohol consumption, tumor size, differentiation degree and lymph node metastasis. The expression level of CCT-α protein was significantly increased in patients with a history of smoking and alcohol consumption (P=0.001, P=0.004). With the increase of tumor diameter, the expression of CCT-α protein was significantly increased (P=0.005). According to histopathological grade, the lower the degree of tumor differentiation, the higher the expression level of CCT-α protein (P=0.000). The expression of CCT-α protein was significantly higher in patients with lymph node metastasis compared with no lymph node metastasis (P=0.000). Quantitative real-time PCR results showed the CCT-α mRNA expression level was significantly higher in OSCC cells than that in normal oral epithelial cells (P=0.016). The protein expression level of CCT-α was significantly higher in OSCC cells than that in normal oral epithelial cells. Conclusions: CCT-α may play a critical role in the carcinogenesis and development of OSCC.

[Endoscopy-assisted sialodochoplasty for the treatment of severe sialoduct stenosis].

OBJECTIVE: To evaluate the effects of endoscopy-assisted sialodochoplasty for the treatment of severe sialoduct stenosis with concurrent megaducts. METHODS: From Jul.2010 to Dec. 2016, 8 patients presenting with severe parotid duct stenosis and 3 patients with occlusion of the Wharton's duct underwent endoscopy-assisted sialodochoplasty.All these patients had concurrent severe ductal ectasiaand manifested a painful swelling of the involved salivary glands.The diameter of ectasia and length of stenosis of the sialoducts were measured preoperatively by sialography, computed tomography, or ultrasonography. The megaducts were opened transorally and sutured to the buccal or oral floor mucosa, therefore creating a neo-ostium. All the patients were followed up periodically after operation. The treatment effects were evaluated by clinical signs, sialogram and sialometry. RESULTS: The length of the Stensen's duct stenosis was 5-12 mm, and the diameter of the concurrent ectasia was 8-16 mm. The length of the Wharton's duct stenosis was 10-20 mm, and the diameter of the concurrent ectasia was 6-8 mm.The neo-ostiums healed uneventfully 2 weeks after operation. The duration of the follow-up varied from 6 to 78 months (median: 24 months). Among the 8 patients with Stensen's duct stenosis, two experienced re-obliteration of the neo-ostium, but the buccal bulge and clinical symptoms disappeared; one reported recurrent clinical symptoms after initial alleviation, which could be controlled with self-massaging; the remaining 5 patients had satisfactory clinical results, i.e., disappearance of the obstruction symptoms and buccal bulge, patent ostium,clean saliva and improvement of the ductal ectasia on sialogram. Three patients with Wharton's duct occlusion were asymptomatic with clear saliva and patent ostium;two exhibited approximately normal appearance and one showed improvement of the sialogram.Sialometry was performed in 9 patients with patent neo-ostium of the involved glands,the resting saliva flow rate of the affected glands showed no differences compared with the normal side, and stimulated flow rate showed a significant increase, though less than the control side.The clinical results included good in 5 patients, fair in 4 patients, and poor in 2 patients, with a total effective rate of 82% (9/11). CONCLUSION: Endoscopy-assisted sialodochoplasty appears to be effective and can be a viable option for patients presenting with severe sialoducts tenosis and concurrent ectasia.

[Influence of phosphoglycerate kinase 1 on metastasis and invasion of hepatoma cells and prognosis of liver cancer].

Objective: To investigate the association between expression of phosphoglycerate kinase 1 (PGK1) in liver cancer tissue and prognosis, as well as its influence on metastasis and invasion of hepatocellular carcinoma (HCC) cells. Methods: Overexpression and downregulated expression of PGK1 in HCC cells were mediated by lentivirus to establish hepatoma cell lines with different expression levels of PGK1. The Transwell chamber invasion assay, wound healing assay, and colony-forming assay were used to investigate the influence of PGK1 on metastasis, invasion, and proliferation of HCC cells. Immunohistochemistry was used to measure the expression of PGK1 in liver cancer tissue samples from 116 patients with hepatocellular carcinoma who underwent radical surgery, and the Kaplan-Meier method and the log-rank test were used to determine the association between PGK1 expression and prognosis of patients with liver cancer. Results: HCCLM3 and MHCC97H HCC cells with high metastatic potential had significantly higher expression of PGK1 than Hep3B and Huh7 HCC cells with low metastatic potential. Downregulation of PGK1 expression significantly inhibited the migration (31.2% ± 2.4% vs 12.0% ± 1.3%, t = 21.57, P < 0.01), invasion (58 ± 11 vs 21 ± 8, t = 4.687, P < 0.05), and colony-forming ability (168.6 ± 15.1 vs 118.4 ± 8.1, t = 6.650, P < 0.05) of MHCC97H cells, while overexpression of PGK1 enhanced the migration (62.8% ± 4.4% vs 83.6% ± 6.1%, t = 20.56, P < 0.01), invasion (80 ± 12 vs 121 ± 15, t = 4.603, P < 0.05), and colony-forming ability (52.3 ± 8.6 vs 84.7 ± 9.0, t = 27.18, P < 0.01) of Hep3B cells. The high PGK1 expression group had significantly shorter median disease-free survival time and mean survival time than the low PGK1 expression group (22.00 ± 8.51 vs not reached, P < 0.05; 46.00 ± 16.87 vs not reached, P < 0.01). Conclusions: PGK1 is involved in the regulation of metastasis and invasion of HCC cells and can promote the migration and invasion of HCC cells. Therefore, PGK1 may be an important predictor of prognosis and postoperative recurrence in patients with liver cancer.

[Mononuclear cells of umbilical cord blood differentiation to granulocyte cell in vitro].

Objective: To explore an optimal method for granulocyte cell production from umbilical cord blood mononuclear cells. Methods: Erythrocytes were precipitated by hydroxyethyl starch. Mononuclear cells were isolated through Ficoll density gradient centrifugation. Different media, additives and cultivation model were chosen for granulocyte induction. Cell morphology was observed by microscopy, and cell phenotype was detected by flow cytometry. The CD18 expression of granulocytes was tested by immunofluorescence assay, and phagocytosis test was executed as well. Results: Compared to fetal bovine serum (FBS) treatment group, cell viability, counts and differentiation rate of granulocytes induced by X-VIVO(TM) 15 combined with TPO, SCF, G-CSF but without FBS were superior. And X-VIVO(TM)15 medium was better than SCGM medium at effectiveness and cost. Using two-stage mode of hematopoietic stem cell expansion followed by granulocyte induction with X-VIVO(TM)15 combining TPO, SCF and G-CSF, cell proliferation was nearly 132 times at day 21. Flow cytometry showed that the differentiation was lagged in 2-stage mode than in direct induction mode, CD15 expression was (69.60± 1.06) % vs (97.73±0.39) %; Wright-Giemsa staining demonstrated mature granulocytes; immunofluorescence showed the expression of lysosomal proteins CD18. A strong phagocytic function of mature granulocytes was demonstrated by phagotrophic efficiency of (51.43±0.05) %. And granulocyte had chemotaxis ability under the role of chemotactic factor IL-8. Conclusion: Optimized culture media and cultivation mode are achieved for functional granulocytes induction in vitro.

Differential diagnosis of IgG4-related sialadenitis, primary Sjögren syndrome, and chronic obstructive submandibular sialadenitis.

Our aim was to differentiate IgG4-related sialadenitis, primary Sjögren syndrome, and chronic obstructive submandibular sialadenitis by analysing clinical, radiographic, and pathological features. Fifty-five patients, 50, and 50 were enrolled, respectively and their baseline characteristics and serological, sialographic, and pathological findings compared. The male:female ratio for IgG4-related sialadenitis was 1:1.2 for primary Sjögren syndrome 1:15.7, and for chronic obstructive submandibular sialadenitis1:0.92. Numbers with enlarged salivary glands were 55, 16, and 50; with xerostomia 26, 48, and 0; with a history of allergy 26, 4, and 6, and with coexisting systemic disease 12, 19, and 0 (p=0.14). Mean (SD) serum IgG4 concentrations were 109.1 (97.9), 4.9. (1.9) g/L, and 5.3 (1.6) g/L, p<0.001 in all cases. Sialography showed enlargement of the gland, dilatation of the duct, and slightly decreased secretory function in IgG4-related disease; obvious sialectasia and decreased secretory function in Sjögren syndrome; and dilatation of Wharton's duct and filling defects in obstructive sialadenitis. Histopathological examination showed lymphoplasmacytic infiltration with storiform fibrosis, lymphoplasmacytic inflammation and lymphoepithelial lesions, and dilatation of the duct with epithelial metaplasia in the three groups, respectively. The number of IgG4-positive plasma cells was 123 (45)/HPF, 8 (3)/HPF, and 5 (4)/HPF, while the IgG4-/IgG-positive cell ratio was 71.7 (13.9)%, 4.6 (2.5)%, 18.9 (19.7)%, respectively (p<0.001). The three conditions have different clinical, radiographic, and pathological features that provide important clues to the differential diagnosis. Serological and histological tests are important, and comprehensive consideration is necessary.