Objective:To investigate the dynamic characteristics of human papillomavirus (HPV) genomic integration during cervical lesion progression and the clinical value of HPV integration detection in stratify HPV-positive women, and to explore its molecular mechanisms in cervical carcinogenesis.Methods:A prospective cohort study was designed to enroll high-risk HPV (HR-HPV) positive women who underwent cervical cancer screening in Drum Tower Hospital Affiliated to Nanjing University Medical School and Nanjing Maternity and Child Health Care Hospital from July 2022 to July 2024. Cervical exfoliated cells samples were collected, and HPV whole genome targeted capture and high-throughput sequencing technology were used. The HPV integration patterns, host gene functional region distribution and pathway enrichment characteristics of 157 samples with different cervical lesions grades were analyzed, including 31 cases of normal cervix, 40 cases of cervical intraepithelial neoplasia (CIN) Ⅰ, 32 cases of CIN Ⅱ, 42 cases of CIN Ⅲ, and 12 cases of cervical cancer.Results:HR-HPV integration was detected in 80.2% (126/157) of the 157 HR-HPV positive samples. The incidence of HR-HPV integration in cervical cancer patients was 12/12, which was higher than that in normal women (77%, 24/31). The incidence of HPV16 integration was significantly higher in high-grade lesions, and the incidence of HPV16 integration was 43% (18/42) in CIN Ⅲ patients and 8/12 in cervical cancer patients ( P<0.001). A total of 14 438 integration events were detected in 126 samples with HPV integration. The integration sites were mainly distributed in the host intergenic region (51.0%, 7 359/14 438) and intronic region (38.1%, 5 494/14 438), and the integration frequency of viral L1 gene was the highest (28.4%, 4 498/16 781). Functional enrichment analysis showed that HPV integration-related host genes were significantly enriched in transport of small molecules,cyclic guanosine monophosphate (cGMP)-protein kinase G (PKG) signaling pathway, and purine ribonucleotide biosynthetic process, which synergistically drove carcinogenesis through multiple mechanisms. Conclusions:HPV integration events are significantly associated with the progression of cervical lesions. HPV integrated detection based on cervical exfoliated cells is expected to optimize the current screening strategy, reduce excessive intervention of HPV positive women and facilitate their accurate triage management.

Objective:To investigate the dynamic characteristics of human papillomavirus (HPV) genomic integration during cervical lesion progression and the clinical value of HPV integration detection in stratify HPV-positive women, and to explore its molecular mechanisms in cervical carcinogenesis.Methods:A prospective cohort study was designed to enroll high-risk HPV (HR-HPV) positive women who underwent cervical cancer screening in Drum Tower Hospital Affiliated to Nanjing University Medical School and Nanjing Maternity and Child Health Care Hospital from July 2022 to July 2024. Cervical exfoliated cells samples were collected, and HPV whole genome targeted capture and high-throughput sequencing technology were used. The HPV integration patterns, host gene functional region distribution and pathway enrichment characteristics of 157 samples with different cervical lesions grades were analyzed, including 31 cases of normal cervix, 40 cases of cervical intraepithelial neoplasia (CIN) Ⅰ, 32 cases of CIN Ⅱ, 42 cases of CIN Ⅲ, and 12 cases of cervical cancer.Results:HR-HPV integration was detected in 80.2% (126/157) of the 157 HR-HPV positive samples. The incidence of HR-HPV integration in cervical cancer patients was 12/12, which was higher than that in normal women (77%, 24/31). The incidence of HPV16 integration was significantly higher in high-grade lesions, and the incidence of HPV16 integration was 43% (18/42) in CIN Ⅲ patients and 8/12 in cervical cancer patients ( P<0.001). A total of 14 438 integration events were detected in 126 samples with HPV integration. The integration sites were mainly distributed in the host intergenic region (51.0%, 7 359/14 438) and intronic region (38.1%, 5 494/14 438), and the integration frequency of viral L1 gene was the highest (28.4%, 4 498/16 781). Functional enrichment analysis showed that HPV integration-related host genes were significantly enriched in transport of small molecules,cyclic guanosine monophosphate (cGMP)-protein kinase G (PKG) signaling pathway, and purine ribonucleotide biosynthetic process, which synergistically drove carcinogenesis through multiple mechanisms. Conclusions:HPV integration events are significantly associated with the progression of cervical lesions. HPV integrated detection based on cervical exfoliated cells is expected to optimize the current screening strategy, reduce excessive intervention of HPV positive women and facilitate their accurate triage management.

Objective To investigate the current status of liver and gallbladder diseases and analyze the influencing factors of fatty liver based on the ultrasound examination results of officers and soldiers on a certain island.Methods The data of physical examination from 377 officers and soldiers on a certain island in 2021 were retrospectively analyzed.Age,gender,living habits,abdominal ultrasound results,and liver function test results were collected,and the incidence of liver and gallbladder diseases were investigated.Univariate and multivariate logistic regression analyses were used to screen the influencing factors of fatty liver disease.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of influencing factors for fatty liver disease.Results A total of 103 officers and soldiers were diagnosed with liver and gallbladder diseases.The top three were fatty liver(63.11%,65/103),gallbladder polyps(19.42%,20/103),and liver hemangioma(8.74%,9/103),accounting for 91.27%(94/103)of the subjects with liver and gallbladder diseases and 24.93%(94/377)of all the subjects undergoing physical examination.There were significant differences in the age,body mass index(BMI),the habit of mid night snacks,liver function indexes(alanine aminotransferase[ALT],aspartate aminotransferase[AST],total bilirubin[T-Bil],direct bilirubin[D-Bil],gamma-glutamyltransferase[γ-GT]),glucose(Glu),and triglyceride(TG)between the officers and soldiers with fatty liver(65 cases)and without fatty liver(312 cases)(all P<0.01).Multivariate logistic regression analysis showed that ALT,Glu,TG,BMI and the habit of midnight snacks were independent influencing factors of fatty liver(all P<0.05).The area under the ROC curve values of ALT,Glu,TG,BMI,the habit of midnight snacks and their combination in prediction of fatty liver were 0.776,0.621,0.764,0.828,0.637 and 0.866,respectively.Conclusion There is a high prevalence of fatty liver in officers and soldiers on a certain island(17.24%,65/377).ALT,Glu,TG,BMI and the habit of midnight snack can be used as a combined diagnostic model of fatty liver.The diet,exercise and training methods should be systematically and comprehensively managed to control related risk factors.Risk factor intervention and treatment should be carried out for high-risk population to effectively improve the physical and mental health of officers and soldiers on the island.

Objective:To explore the feasibility and clinical effectiveness of preoperative positioning grid combined with CTA assisted design of anterolateral thigh perforator flap (ALTPF) in reconstruction of soft tissue defects around foot and ankle.Methods:From May 2018 to December 2021, a total of 18 cases with soft tissue defects around foot and ankle were admitted to the Department of Plastic and Reconstructive Surgery, First Affiliated Hospital of Ningbo University. The patients were 11 males and 7 females, aged from 32 to 78 years old, with an average age of 57.5 years old. Among them, 10 had chronic ulcer wounds, 6 had traumatic wounds, and 2 had postoperative wounds after malignant tumor resection. The sizes of soft tissue defect ranged from 6.0 cm×8.0 cm to 9.0 cm×13.0 cm, and the flap sizes ranged from 8.0 cm×10.0 cm to 11.0 cm×15.0 cm. Preoperative positioning grid combined with CTA three-dimensional reconstruction were used to mark the exit point of the perforator vessels in digital format, in order to restore the course of vessels and calculate the length of the vascular pedicles. ALTPFs were accurately designed based on the digitally reconstructed images, and then the ALTPFs were used to repair the soft tissue defects around the ankle. The flap donor sites were directly closed in stage Ⅰ. After the operation, all the patients were included in scheduled follow-ups at the outpatient department to observe the appearance of the recipient flaps and donor sites. The functional evaluation of the affected feet were assessed according to the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score.Results:All 18 flaps survived, and the wounds in both the recipient and donor sites healed in stageⅠ. Postoperative follow-up for the 18 patients were 5 to 36 months, with an average of 13 months. At the last follow-up, the appearances of the flaps were good without swelling, the donor sites had good recovery, and there was no obvious scar hypertrophy. The function evaluation of the affected feet were found at excellent in 10 cases, good in 6 cases, and fair in 2 cases.Conclusion:Preoperative position grid combined with CTA three-dimensional reconstruction can digitally mark the exit points and running courses of the perforator vessels. It is an effective method for accurate position of perforator vessels before surgery. It can effectively reduce the operation time, lower the surgical risks, and achieve a high survival rate of the flap, thus holding considerable clinical value.

Objective:To investigate the therapeutic effect of conjoint fascia sheath suspension and levator palpebrae muscle shortening on congenital severe blepharoptosis.Methods:From June 2014 to December 2018, 30 cases (40 eyes) of congenital severe ptosis were treated in Ningbo First Hospital and Shaoxing Women and Children＇s Hospital. All patients were corrected by conjoint fascia sheath suspension and levator palpebrae muscle shortening. Six months after operation, the distance between the middle point of upper eyelid margin and corneal reflex point was measured to evaluate the correction effect of blepharoptosis; the improvement of upper eyelid appearance was evaluated by 5-point Likert scale (LS), and the incidence of complications was counted.Results:Thirty patients (40 eyes) were followed up for 6-12 months. Correction effect showed that preoperative marginal reflex distance (MRD) was (0.10±0.05) and postoperative MRD was (3.80±0.55); the difference was statistically significant ( t=0.95, P<0.05); 38 eyes (95%) were corrected and 2 eyes (5%) were undercorrected; appearance of upper eyelid showed that preoperative LS was (0.50±0.05) and postoperative LS (3.80±0.55); the difference was statistically significant ( t=0.98, P<0.05). Only one case was complicated with keratitis caused by improper early nursing after incomplete closure exposure, and the patient was cured by sealing the eyes combined with drug conservative treatment. Conclusions:Combined conjoint fascial sheath suspension and levator palpebrae shortening in the treatment of severe blepharoptosis has significant effect, with the advantages of simple operation, small surgical trauma, low recurrence rate, low incidence of complications, high predictability of surgical results and high satisfactory rate of patients.

Objective:To analyze the factors influencing prognosis of intrahepatic cholangiocarcinoma (ICC) after surgical resection.Methods:The clinical data of patients diagnosed with ICC and who underwent surgical resection from December 2015 to December 2019 at the Fifth Medical Center of PLA General Hospital were retrospectively analyzed. Of 39 patients who were included in this study, there were 23 males and 16 females, with age of (54.1±7.2) years old. The body mass index, hepatitis B virus infection status, tumor diameter, degree of differentiation, microvascular tumor thrombus, lymph node metastasis, and serum levels of carbohydrate antigen 19-9 (CA19-9) were analyzed as risk factors affecting postoperative recurrence and survival.Results:The median times to recurrence were significantly better in patients with a tumour length <5 cm (11 vs. 5 months), patients without microvascular tumor thrombus (54 vs. 6 months) and patients without lymph node metastasis (8 vs. 5 months) (all P<0.05). The median survival of patients with CA19-9≥100 U/ml was significantly shorter than that of patients with CA19-9<100 U/ml, (9 vs. 27 months, P<0.05). Tumor diameter>5 cm, microvascular tumor thrombus, lymph node metastasis, and CA19-9 ≥100 U/ml are risk factors affecting the recurrence time after ICC resection, CA19-9 ≥100 U/ml is a risk factor affecting survival time after ICC resection. Conclusion:Tumor diameter, microvascular tumor thrombus, lymph node metastasis and CA19-9 can be used to estimate the risk of ICC recurrence, and CA19-9 level can be used to estimate postoperative survival of ICC patients after resection.

Objective: To investigate the clinical effect of free medical sural artery perforator myocutaneous flap for repairingof the woundwith osteomyelitis. Methods: 17 patients suffered from the wound with osteomyelitis were treated in the Ningbo First People′s Hospital, There were 11 males and 6 females with an mean age of 53.2 years (range, 21-76 years). The sizes of the defect ranged 5 cm×4 cm to 13 cm×6 cm. All patients underwent debridement and used antibiotic-loaded bonecement to cover the wound. Meanwhile, patients were treated with sensitive antibiotics, operation and free medical sural artery perforator myocutaneous flap were used to treat the wound. Preoperative use ultrasound and CT angiography to positioning perforator, The flap area ranged from 6 cm×4 cm to 13 cm×7 cm and the donor sites were closed directly. The author provided the patients with the treatment of anti-inflammatory, anti-spasmodic and anti-coagulantin the postoperative. Used infrared thermograms to assess the flap blood supply. Results: One flap skin margin was non union due to poor blood supply.All of the other 16 flaps success survived and the donor sites were closed directly. Postoperative follow-up period was 4 to 23 months and the flaps had satisfied texture and appearance.All the donor sites had a good healing with no pain and complications, also the osteomyelitis was controlled. Conclusions The free medical sural artery perforator myocutaneous flap is reliable for reconstruction of the wound with osteomyelitis.

Objective: To investigate the clinical effect of reconstruction of sensory anterolateral thigh perforator flaps in repairing soft tissue defects after radical operation for Paget disease of perineum. Methods: From April 2017 to July 2017, Ningbo First Hospital treated 3 male patients with Paget′s disease of perineum, 2 patients with RayA2 stage and 1 patient with RayB stage. The range of soft tissue defect after resection of tumors is 8 cm×10 cm to 10 cm×12 cm. The anterolateral thigh perforator flaps were designed to repair skin defect, and the lateral thigh cutaneous nerve branch was contained to reconstruct protective sensation and restore autonomic nerve function. Tension-free direct suture was performed in part of the donor site, of which 3 cases were difficult to suture directly, 2 cases were treated with skin grafting and 1 case was closed with skin distractor. Results: All 3 flaps survived. The average area of the flaps was about 12 cm×14 cm. The average follow-up period was 3 months. The appearance and function of the flaps were satisfactory. The average sensory recovery of the flaps was as high as S3. The autonomic nervous function was partially restored. Some of the skin had sweating function and normal skin temperature. At the same time, there was no difference between the urination function and sexual competence before operation. The donor site wound healed well and its appearance and function recovered well. Conclusions The reconstruction of sensory anterolateral thigh perforator flaps is an good method for repairing soft tissue defect after radical resection of Paget disease of perineum. It has satisfactory results in the protection and reconstruction of donor and recipient areas.

To investigate the clinical efficacy of one-stage repairing the wounds after excision of soft tissue malignant tumor in lower limb with deep inferior epigastric perforator (DIEP) flaps. Methods From Au-gust, 2016 to July, 2018, 11 patients(8 males and 3 females; aged 35-76 years) with the soft tissue defects in the low-er limb underwent immediate reconstructive operations with DIEP flap after their tumor resection. The defects ranged in area from 11 cm×7 cm to 25 cm×10 cm after extended resection, including 5 cases of bone exposure. The DIEP flap area ranged from 11 cm×8 cm to 26 cm×18 cm.Among them, 9 cases were treated with free transplantation and 2 cases with pedicle transplantation.The donor sites were closed directly. The patients’ limb function, tumor recurrence and the healing of surgical incision had been followed-up. Results Eleven flaps survived and the donor sites healed directly. One flap had skin pigmentation due to postoperative vascular crisis, and the other flaps had satisfied texture and appearance. All patients were followed-up from 6 to 30 months. The lower limb movement was normal. All the donor sites had a good healing with no pain and abdominal complications. No tumor recurrence was observed during the followed-up period. Conclusion DIEP flap have many advantages, such as large cutting area, which is suitable for large-area wound repair after extended excision of soft tissue malignant tumor in lower limb. At the same time, the perforator vessel is constant and the caliber are thick, also the donor area is concealed and can be sutured directly.This kind of the technique for reconstruction of lower limb soft tissue defects after malignant tumor resection is well acceptable.

Objective To explore the method and clinical efficacy of microsurgical reconstructive surgery af-ter malignant tumors radical resection on the surface. Methods From June, 2015 to March, 2017, a total of 32 pa-tients in this group underwent radical resection of malignant tumors. All of them resulted in a large area of soft tissue defect with loss of function.The average defect area was 10.0 cm×12.0 cm.All types of free flap and pedicle flap were selected. Myocutaneous flaps or local flaps were used to repair large areas of soft tissue defect on the basis of micro-surgical techniques to protect the blood supply of the flaps and to reduce the damage to the donor area by evaluating the defect size, surrounding anatomical characteristics, and other factors. Results Thirty-two flaps survived com-pletely. After 4 to 18 months of followed-up, the color of flap was close to that of the surrounding skin. The texture was good. There was no bloated swelling, and the skin protective sensation was restored. There was no obvious scar hyperplasia, no chills, no pain and no other complications in the donor site. Conclusion After the radical mastec-tomy of the body surface, different types of flaps can be selected for the first-stage microsurgery according to the characteristics of the different parts and the tumor excision wounds to obtain a good clinical efficacy.