[Q-value-guided femtosecond laser-assisted in situ keratomileusis for low-to-moderate myopia with age-related accommodation deficiency].

Objective: To evaluate clinical outcomes and visual quality after femtosecond laser-assisted in situ keratomileusis (FS-LASIK) performed with the Q-value-guided optimized monocular vision protocol (Custom-Q) for correction of low-to-moderate myopia and compensation for age-related accommodation deficiency. Methods: A prospective study was performed based on patients with myopia and age-related accommodation deficiency, who underwent Custom-Q FS-LASIK in the Peking University Third Hospital from October 2022 to October 2023. Monocular and binocular distance, intermediate, and near visual acuities, simulated whole-course visual acuity (binocular defocus curve), objective and subjective refractions, anterior corneal Q factor, anterior corneal higher-order aberrations and a subjective questionnaire assessing near visual acuity and visual quality were evaluated at 3 months postoperatively. Visual acuity was recorded using the logarithm of the minimum angle of resolution. The data were analyzed using the t-test and the Wilcoxon rank-sum test. Results: A total of 45 patients (90 eyes) who met the inclusion criteria and completed the 3-month follow-up were included in the study. The mean age of the 21 men (42 eyes) and 24 women (48 eyes) was (42.82±2.72) years. During the 3-month follow-up, all the patients achieved binocular uncorrected distance visual acuity not less than 0.00 and uncorrected near visual acuity not less than 0.20. Defocus curves revealed better results at 3 months postoperatively [0.00 (-0.08, 0.00), -0.08 (-0.08, 0.00)] at intermediate and near vergence (-1.50 to -1.00 D) compared to the preoperative values [0.00 (-0.08, 0.05), -0.08 (-0.08, 0.00)] (P<0.05). All the 45 patients maintained 0.10 or better vision at the defocus range from -2.50 to 0.00 D. The Q value in the dominant eyes was more positive postoperatively (0.01±0.26 vs.-0.15±0.14, P<0.05), while in the nondominant eyes, the Q value and corneal spherical aberration coefficient became more negative than those before surgery (-0.27±0.32 vs.-0.14±0.11, P<0.05; 0.08±0.05 vs. 0.12±0.03, P<0.05). The questionnaire demonstrated 42 (93.3%) patients were satisfied with near vision. The three most commonly reported visual discomforts after surgery were blurred vision, glare and halos. These symptoms were all mild to moderate in severity and did not interfere with daily life. Conclusion: For myopic patients with age-related accommodation deficiency, the Custom-Q algorithm proved to be a safe and effective way to achieve acceptable near vision and visual quality without compromising distance vision.

The effect of small molecule inhibitor NSC348884 on nucleophosmin 1-mutated acute myeloid leukemia cells.

OBJECTIVE: Nucleophosmin 1 (NPM1) is a common shuttling protein. Mutation in the NPM1 gene is the most frequent gene alteration in acute myeloid leukemia (AML). This study aims to explore the inhibitory effects of small molecule NSC348884 on wild-type and NPM1-mutated AML cells. MATERIALS AND METHODS: Immunofluorescence was used to determine the intracellular localization of NPM1 protein in wild-type (OCI-AML2) and NPM1-mutated (OCI-AML3) AML cell lines. The oligomerization state of NPM1 was assessed by Western blot analysis, and the inhibitory effect of NSC348884 on the proliferation of AML cells was evaluated by Cell-counting kit-8 (CCK-8). Flow cytometry was used to detect the proapoptotic effect of NSC348884 on AML cells. RESULTS: Western blot results showed a significant reduction in the levels of the oligomeric NPM1 protein after the treatment with NSC348884. NSC348884 had an inhibitory effect on the proliferation of both wild-type and NPM1-mutant AML cells. The inhibitory effect on OCI-AML3 cells was stronger, compared to OCI-AML2 cells. Flow cytometry showed that NSC348884 could significantly induce AML cell apoptosis and had a stronger proapoptotic effect on OCI-AML3 cells. CONCLUSIONS: NSC348884 had inhibitory and proapoptotic effects on both wild-type and NPM1-mutated AML cells. The effect of NSC348884 on AML cells, carrying NPM1 mutation was significantly stronger.

[Comprehensive assessment of mismatch repair and microsatellite instability status in molecular classification of endometrial carcinoma].

Objective: To explore the concordance and causes of different mismatch repair (MMR) and microsatellite instability (MSI) detection results in endometrial carcinoma (EC) molecular typing. Methods: A total of 214 EC patients diagnosed from January 2021 to April 2023 were selected at the Department of Pathology, Peking University Third Hospital. The immunohistochemistry (IHC) results of MMR protein were reviewed. Tumor specific somatic mutations, MMR germline mutations, microsatellite scores and tumor mutation burden (TMB) were detected by next-generation sequencing (NGS) with multi-gene panel. Methylation-specific PCR was used to detect the methylation status of MLH1 gene promoter in cases with deficient MLH1 protein expression. In cases with discrepant results between MMR-IHC and MSI-NGS, the MSI status was detected again by PCR (MSI-PCR), and the molecular typing was determined by combining the results of TMB and MLH1 gene promoter methylation. Results: (1) In this study, there were 22 cases of POLE gene mutation subtype, 55 cases of mismatch repair deficient (MMR-d) subtype, 29 cases of p53 abnormal subtype, and 108 cases of no specific molecular profile (NSMP). The median age at diagnosis of MMR-d subtype (54 years old) and the proportion of aggressive histological types (40.0%, 22/55) were higher than those of NSMP subtype [50 years old and 12.0% (13/108) respectively; all P<0.05]. (2) Among 214 patients, MMR-IHC test showed that 153 patients were mismatch repair proficient (MMR-p), 49 patients were MMR-d, and 12 patients were difficult to evaluate directly. MSI-NGS showed that 164 patients were microsatellite stable (MSS; equal to MMR-p), 48 patients were high microsatellite instability (MSI-H; equal to MMR-d), and 2 patients had no MSI-NGS results because the effective sequencing depth did not meet the quality control. The overall concordance between MMR-IHC and MSI-NGS was 94.3% (200/212). All the 12 discrepant cases were MMR-d or subclonal loss of MMR protein by IHC, but MSS by NGS. Among them, 10 cases were loss or subclonal loss of MLH1 and (or) PMS2 protein. Three discrepant cases were classified as POLE gene mutation subtype. In the remaining 9 cases, 5 cases and 3 cases were confirmed as MSI-H and low microsatellite instability (MSI-L) respectively by MSI-PCR, 6 cases were detected as MLH1 gene promoter methylation and 7 cases demonstrated high TMB (>10 mutations/Mb). These 9 cases were classified as MMR-d EC. (3) Lynch syndrome was diagnosed in 27.3% (15/55) of all 55 MMR-d EC cases, and the TMB of EC with MSH2 and (or) MSH6 protein loss or associated with Lynch syndrome [(71.0±26.2) and (71.5±20.1) mutations/Mb respectively] were significantly higher than those of EC with MLH1 and (or) PMS2 loss or sporadic MMR-d EC [(38.2±19.1) and (41.9±24.3) mutations/Mb respectively, all P<0.01]. The top 10 most frequently mutated genes in MMR-d EC were PTEN (85.5%, 47/55), ARID1A (80.0%, 44/55), PIK3CA (69.1%, 38/55), KMT2B (60.0%, 33/55), CTCF (45.5%, 25/55), RNF43 (40.0%, 22/55), KRAS (36.4%, 20/55), CREBBP (34.5%, 19/55), LRP1B (32.7%, 18/55) and BRCA2 (32.7%, 18/55). Concurrent PTEN, ARID1A and PIK3CA gene mutations were found in 50.9% (28/55) of MMR-d EC patients. Conclusions: The concordance of MMR-IHC and MSI-NGS in EC is relatively high.The discordance in a few MMR-d EC are mostly found in cases with MLH1 and (or) PMS2 protein loss or MMR protein subclonal staining caused by MLH1 gene promoter hypermethylation. In order to provide accurate molecular typing for EC patients, MLH1 gene methylation, MSI-PCR, MMR gene germline mutation and TMB should be combined to comprehensively evaluate MMR and MSI status.

[Correlation between pulmonary quantitative CT measurement indicators and respiratory symptoms in patients with chronic obstructive pulmonary disease in stable stage].

Objective: To explore the correlation between pulmonary quantitative CT measurement indicators and respiratory symptoms in patients with stable chronic obstructive pulmonary disease (COPD). Methods: A total of 186 patients with COPD in stable stage who visited in the outpatient department of Beijing Hospital from March 2021 to February 2022 were prospectively included. Demographic data, respiratory symptoms and lung function were collected. The original DICOM data of high-resolution CT (HRCT) were processed using the FACT medical imaging information system and the pulmonary emphysema index pixel index-950 (PI-950) and the airway wall thickness (4-6 T) and the percentage of airway area (4-6 WA%) of the 4-6 generation bronchi which represent the segmental and subsegmental bronchi were measured automatically. According to the modified British medical research council dyspnea scale (mMRC, 0-1 point for low score group, 2-4 points for high score group), chronic obstructive pulmonary disease assessment test (CAT, score<10 points for low score group,≥10 points for high score group), cough, expectoration and wheezing (asymptomatic group and symptomatic group), they were divided into two groups as dependent variables. The relationship between imaging parameters and the above symptoms was evaluated using a logistic regression model. Results: The study ultimately included 186 patients who met the inclusion criteria, including 162 males and 24 females, aged (68.9±9.3) years old. There were 83 patients in the high mMRC group, 120 patients in the high CAT group, 146 patients in the cough group, 154 patients in the expectoration group, and 65 patients in the wheezing group. The age and emphysema parameter PI-950 in the high score group of mMRC were higher than those in the low score group, while the percentage of the forced expiratory volume in 1 second (FEV1) predicted value (FEV1 pred) after medication, the percentage of carbon monoxide diffusion volume (DLCO) predicted value (DLCO pred), and the percentage of the maximum midexpiratory flow (MMEF) predicted value (MMEF pred) after medication were lower than those in the low score group (all P<0.05). The age of the high CAT group was higher than that of the low score group, while FEV1 pred and MMEF pred after medication were lower than those of the low score group (all P<0.05). The proportion of males, patients with smoking history, and smoking index in the cough group were higher than those in the non cough group, while the 4 WA% was lower than that in the non cough group (all P<0.05). The proportion of males, patients with smoking history, smoking index, and PI-950 in the expectoration group were higher than those in the non expectoration group, while FEV1 pred after medication and 4 WA% were lower than those in the non expectoration group (all P<0.05). The 5 WA% and 6 WA% of the wheezing group were higher than those of the non wheezing group, while MMEF pred after medication was lower than that of the non wheezing group (all P<0.05). Multivariate logistic regression analysis showed that after adjusting for demographic characteristics, smoking, combined diseases, lung function and other confounding factors, for every 10% increase in PI-950, the likelihood of developing more severe dyspnea for the patients (high score group according to mMRC) increased by 67.3% (OR=1.673, 95%CI: 1.052-2.658); Every 10% increase in 6WA% increased the likelihood of wheezing by 3.189 times (OR=4.189, 95%CI: 1.070-16.395). No correlation was found between various imaging indicators and cough, expectoration, and CAT scores (P>0.05). Conclusion: Quantitative CT measurement indicators in stable COPD patients can explain the presence and severity of respiratory symptoms, the pulmonary emphysema indicator is associated with dyspnea, and the percentage of proximal airway wall area is associated with wheezing.

[Malignant gastrointestinal neuroectodermal tumor: a clinicopathological analysis of three cases].

Objective: To investigate the clinicopathological characteristics of malignant gastrointestinal neuroectodermal tumors (GNET), and to describe their clinical, histological, immunophenotypic, ultrastructural, and molecular features, diagnosis and differential diagnosis. Methods: Three cases of malignant GNET were collected at Xijing Hospital of the Fourth Military Medical University, from 2013 to 2022. All patients underwent surgical resection of the tumor. Histological, immunohistochemical (IHC), ultrastructural and molecular genetic analyses were performed, and the patients were followed up for six months, three years and five years. Results: There were two males and one female patients. The tumors were located in the ileum, descending colon, and rectum, respectively. Grossly, the tumors were solid, firm, and poorly circumscribed, measured in size from 2 to 4 cm in greatest dimension, and had a greyish-white cut surface. These tumors were histologically characterized by a sheet-like or nested population of oval to spindled cells or epithelioid cells with weakly eosinophilic or clear cytoplasm, small nucleoli and scattered mitoses. Electron microscopy showed neuroendocrine differentiation, and no evidence of melanogenesis. IHC staining showed that the tumor cells were diffusely positive for S-100 protein, SOX10, CD56, synaptophysin and vimentin. They were negative for melanocytic markers, HMB45 and Melan A. All three cases showed split EWSR1 signals consistent with a chromosomal translocation involving EWSR1. Next-generation sequencing in one case confirmed the presence of EWSR1-ATF1 fusion. These patients were followed up for 6 months, 3 years and 5 years, respectively, and all of them developed possible lung or liver metastases, and one of them died of multiple pulmonary metastases. Conclusion: Malignant GNET has distinctive morphological, IHC, and molecular genetic features and it should be differentiated from other malignancies of the gastrointestinal tract, especially clear cell sarcoma and melanoma.

Functional identification of ANR genes in apple (Malus halliana) that reduce saline-alkali stress tolerance.

As one of the major abiotic stresses restricting the development of global agriculture, saline-alkali stress causes osmotic stress, ion poisoning, ROS damage and high pH damage, which seriously restrict sustainable development of fruit industry. Therefore, it is essential to develop and cultivate saline-alkali-resistant apple rootstocks to improve the yield and quality of apples in China. Based on transcriptome data, MhANR (LOC114827797), which is significantly induced by saline-alkali stress, was cloned from Malus halliana. The physicochemical properties, evolutionary relationships and cis-acting elements were analysed. Subsequently, the tolerance of MhANR overexpression in Arabidopsis thaliana, tobacco, and apple calli to saline-alkali stress was verified through genetic transformation. Transgenic plants contained less Chl a, Chl b and proline, SOD, POD and CAT activity, and higher relative electrical conductivity (REC) compared to WT plants under saline-alkali stress. In addition, expression of saline-alkali stress-related genes in overexpressed apple calli were also lower than in WT calli, including the antioxidant genes (MhSOD and MhCAT^), the Na+ transporter genes (MhCAX5, MhCAX5, MhSOS1, MhALT1), and the H+ -ATPase genes (MhAHA2 and MhAHA8), while expression of the K+ transporter genes (MhSKOR and MhNHX4) were higher. Expression of MhANR reduced tolerance of A. thaliana, tobacco, and apple calli to saline-alkali stress by regulating osmoregulatory substances, chlorophyll content, antioxidant enzyme activity, and expression of saline-alkali stress-related genes. This research provides a theoretical basis for cultivating apple rootstocks with effective saline-alkali stress tolerance.

[Progress in minimally invasive surgery for adjacent segment disease after lumbar fusion].

Adjacent segment disease (ASDis) is a common complication of posterior lumbar spine fusion and often requires surgical treatment. In the treatment of ASDis, percutaneous spinal endoscopy can be used for simple decompression without removal of the original internal fixation, or for posterior fixation and fusion under the scope or in combination with other access fixation and fusion techniques, with the advantages of less surgical trauma, less bleeding, and faster postoperative recovery. Traditional trajectory screw technique is one of the risk factors for adjacent segment degeneration because of its tendency to cause damage to the adjacent synovial joint during surgery. In contrast, the cortical tone trajectory (CBT) screw placement technique not only reduces the damage to the articular joint during the screw placement process, but also preserves the original internal fixation in the treatment of ASDis, which significantly reduces the surgical trauma. Secondly, the implantation of CBT screws with the aid of digital technologies such as three-dimentinal printed guides, CT navigation, and robotics allows for more precise "double nailing" of ASDis patients to complete the fusion of adjacent segments, and is a minimally invasive procedure to be considered for patients who meet the clinical indications for fusion. This article reviews the literature on the use of percutaneous spinal endoscopy and CBT in the surgical management of ASDis.

[Fumarate hydratase deficient uterine leiomyoma: a clinicopathological and molecular analysis of 80 cases].

Objective: To investigate the clinicopathologic and molecular characteristics of fumarate hydratase (FH) deficient uterine leiomyoma. Methods: Eighty cases of FH deficient uterine leiomyoma were diagnosed from April 2018 to September 2022 in Department of Pathology, Peking University Third Hospital. Sanger sequencing of FH gene exons (exon 1-10) were performed on tumor tissues and matched non-tumor tissues/peripheral blood for all cases. FH immunohistochemistry were performed in 74 cases; S-(2-succino)-cysteine (2SC) were also detected by immunohistochemistry in five cases. Results: Patients' age ranged from 18 to 54 (36.0±7.5) years, with more than 60% exhibiting clinical symptoms of multiple and large leiomyomas (the median diameter was 70 mm). More than four histologic features, including staghorn vasculature, alveolar-pattern edema, bizarre nuclei, oval nuclei arranged in chains, prominent eosinophilic nucleoli with perinucleolar haloes and eosinophilic intracytoplasmic globules were observed in 98.5% (67/68) patients. The immunohistochemical sensitivity of FH and 2SC were 97.3% and 100%, respectively. Based on the Sanger sequencing results, the cases were divided into germline variant group (31 cases), somatic variant group (29 cases) and no variant group (20 cases). Sixty-nine percent (20/29) of the patients with FH germline variation had clear family history. Conclusions: Clinical features, histological morphology, FH and 2SC immunohistochemistry and Sanger sequencing have their own significance and limitations in differential diagnosis of FH deficient uterine leiomyoma. In clinical practice, the above information should be fully integrated and studied for accurate pathologic diagnosis and selection of patients with FH germline variation.

[Activation of JAK/STAT in ovarian high-grade serous cancers and its prognostic significance].

OBJECTIVE: The activation of Janus kinase (JAK) and signal transducers and activators of transcription (STAT) plays an important role in the prognosis and targeted therapy of ovarian high-grade serous carcinoma (HGSC). Utilizing simple and practicable technique, this study aimed to evaluate the activation of JAK/STAT signaling pathway in ovarian HGSC patients, and investigated the correlation between the activation of JAK/STAT signaling pathway and the prognosis of the HGSC patients. METHODS: We performed immunohistochemistry of phosphorylated STAT3 (pSTAT3) and phosphorylated STAT5 (pSTAT5) on paraffin imbedded slides of 73 ovarian HGSC patients, and evaluated the expression level and range of both markers. According to the grading score of the immunostaining of pSTAT3 and pSTAT5, we divided the 73 ovarian HGSC cases into STAT3 low/high expression and STAT5 low/high expression groups, and analyzed the prognosis of the patients in different groups, in order to explore the relationship between the expression of pSTAT3 and pSTAT5 proteins and the prognosis of the HGSC patients. RESULTS: Some of the ovarian HGSC cases showed high expression of pSTAT3 and pSTAT5 protein level, which was related to the poorer prognosis of the HGSC patients. There was a significant difference in the expression level of pSTAT3 and pSTAT5 between the patients with better prognosis (survival time ≥3 years) and poorer prognosis (survival time < 3 years). The patients with higher protein expression of pSTAT3, pSTAT5 or both markers might have poorer prognosis, with significant shorter progression-free survival time and overall survival time (P < 0.001). CONCLUSION: Immunostaining of pSTAT3 and pSTAT5 proteins might be helpful to evaluate and predict the prognosis of the ovarian HGSC patients, and to identify the patients who might have higher chances to respond to the STAT inhibitors and anti-angiogenesis therapy.

[Reclassification of flat type sudden deafness].

Objective: To reclassify the flat type sudden deafness according to the types of audiogram shape, and to explore the correlation between different pattern of hearing loss and prognosis. Methods: All of 1 024 patients with unilateral sudden deafness (492 males and 532 females, aged from 19 to 65 years, with an average age of 41.2 years old) admitted to 33 hospitals nationwide from August 2007 to October 2011 were divided into four types according to Chinese Guideline of Sudden Deafness(2015): low-frequency, high-frequency, flat and total deafness. Then, 402 patients with flat type sudden deafness were further divided into ascending type, descending type and consistent type according to the audiogram shapes. First, we compared the clinical characteristics and prognosis among these three subtypes of flat deafness, then compared the clinical characteristics and prognosis between ascending flat deafness and low-frequency deafness, descending flat deafness and high-frequency deafness, consistent flat deafness and total deafness, explored the factors related to the prognosis of flat deafness. SPSS 21.0 software, ANOVA, χ2 test, t-test and Logistic regression were used to analyze the data. Results: The cure rates of flat ascending, flat descending and flat consistent sudden deafness groups were 70.7%, 17.1% and 34.0% respectively, with a statistically significant difference (χ2=33.984, P<0.001); However, there was no significant difference in age, sex and affected side (all P>0.05). The independent related factors for the recovery of flat type sudden deafness were as follows: whether there was dizziness [OR=0.459; 95% confidence interval (CI): 0.271-0.777], the type of audiogram shape (OR=0.721; 95%CI: 0.530-0.981), and days from onset to therapy (OR=0.903, 95%CI: 0.835-0.978), all of which had P values<0.05. There was no significant difference in the cure rates between ascending flat sudden deafness and low-frequency descending sudden deafness, descending flat sudden deafness and high-frequency descending sudden deafness (all P>0.05). The pure tone average(PTA) of flat consistent sudden deafness and total deafness were (69.1±18.9) and (101.7±17.7) dB HL, respectively, with a statistically significant difference (t=20.890, P<0.001), and the cure rates were 34.0% and 14.5%, respectively, with a statistically significant difference (χ2=29.012, P<0.001). Conclusion: According to the audiogram shape, the flat type sudden deafness can be further divided into ascending flat sudden deafness, descending flat sudden deafness and consistent flat sudden deafness, which can more effectively evaluate the prognosis. The cure rate of ascending flat sudden deafness is similar to that of low-frequency sudden deafness, and the prognosis is well; The cure rate of descending flat sudden deafness is similar to that of high-frequency descending sudden deafness, and the prognosis is poor. The cure rate of consistent flat sudden deafness is higher than that of total deafness. PTA plays an important role in the prognosis of consistent flat sudden deafness and total deafness. Total deafness can be regarded as a single type of sudden deafness.

The impact of instant neutrophil gelatinase-associated lipocalin level on the severity of septic acute kidney injury.

OBJECTIVE: The clinical value of increased levels of neutrophil gelatinase-associated lipocalin (NGAL) in patients with septic acute kidney injury (AKI) is still unclear. This study aimed to assess the link between illness severity and NGAL in patients with septic AKI. PATIENTS AND METHODS: This is a retrospective observational study that took place at the Fourth Hospital of Hebei Medical University, Shijiazhuang, China. The cohort included 365 patients who were admitted to the ICU during the 21-month period. Of them, 18 patients were diagnosed with sepsis (septic group). The average age of patients in the septic group was over 65, and 60.00% of them eventually progressed to septic AKI. Plasma NGAL (pNGAL) and urine NGAL (uNGAL) levels at defined time points were measured. AKI staging was done based on the Kidney Disease Improving Global Outcomes (KDIGO) classification. The Sequential Organ Failure Assessment (SOFA) and Acute Physiology and Chronic Health Evaluation (APACHE) II scores were determined. Patterns and associations between NGAL levels with SOFA scores and different stages of septic AKI were investigated. RESULTS: Both pNGAL and uNGAL showed a positive correlation with SOFA and proved to be reliable predictors of the same. Furthermore, the accuracy of severe sepsis (SOFA ≥ 8) was 0.67 for pNGAL and 0.66 for uNGAL. Real-time detection of pNGAL and uNGAL indicated that they were good biomarkers of severe septic AKI. Area under the receiver operating characteristic (AUROC) for pNGAL and uNGAL were 0.72 (0.69-0.85), and 0.83 (0.71-0.95), respectively. However, only patients with KDIGO 3 AKI presented significantly elevated levels of pNGAL (p < 0.05). Furthermore, the uNGAL level at each stage of septic AKI was higher than that of the non-AKI period (p < 0.01). CONCLUSIONS: In patients with septic AKI, levels of NGAL correlated with SOFA. Levels of pNGAL were good predictors of severe kidney injury and uNGAL levels could detect mild stages of AKI.

[Clinical application of laparoscopic sentinel lymph node mapping in early staged cervical cancer].

Objective: To investigate the application of sentinel lymph node biopsy (SLNB) in early-staged cervical cancer by laparoscopy. Methods: It was a prospective, single-arm, single-center clinical study. Seventy-eight cases of cervical cancer patients were collected from July 2015 to December 2018 at the Fourth Hospital of Hebei Medical University. All the patients were injected with tracer into the disease-free block of cervical tissue after anesthesia by the same surgeon who learned sentinel lymph node (SLN) mapping technique in Memorial Sloan-Kettering Cancer Center, and underwent SLN mapping followed by complete pelvic lymphadenectomy. Moreover, all the dissected lymph nodes were stained with hematoxylin eosin staining (HE) pathological examination. Besides, the negative SLN on hematoxylin-eosin staining were detected by immunohistochemistry cytokeratin staining micro-metastasis. To analyze the distribution, detection rate, false negative rate the sensitivity and negative predictive value of the SLN in early-staged cervical cancer by laparoscopy, and explore the value of SLN mapping in predicting the lymph nodes metastasis in early-staged cervical cancer. Results: The overall detection rate of SLN in cervical cancer was 99% (77/78), bilateral detection rate was 87% (68/78). The average of 12.4 lymph node (LN) and 3.6 SLN were dissected for each patients each side. SLN of cervical cancer were mainly distributed in the obturator space (61.5%, 343/558), followed by external iliac (23.5%, 131/558), common iliac (7.3%, 41/558), para-uterine (3.8%, 21/558), internal iliac (2.2%, 12/558), para abdominal aorta (1.1%, 6/558), and anterior sacral lymphatic drainage area (0.7%, 4/558). Fourteen cases of LN metastasis were found among all 78 cases. There were a total of 38 positive LN, including 26 SLN metastasis and 12 none sentinel LN metastasis. Through immunohistochemical staining and pathological ultra-staging, 1 SLN was found to be isolated tumor cells (ITC), and 5 SLNs were found to be micro-metastases (MIC), accounting for 23% (6/26) of positive SLN. SLN mapping with pathological ultra-staging improved the prediction of LN metastasis in cervical cancer (2/14). Metastatic SLN mainly distributed in the obturator space (65%, 17/26), peri-uterine region (12%, 3/26), common iliac region (15%, 4/26), and external iliac region (8%, 2/26). The consistency of the diagnosis of lymph node metastasis by SLN biopsy and postoperative retroperitoneal lymph node metastasis showed that the Kappa value was 1.000 (P<0.001), indicated that the metastasis status of SLN and retroperitoneal lymph node were completely consistent. The sensitivity, specificity, accuracy, false-negative rate, and negative predictive value of SLN biopsy in the diagnosis of lymph node metastasis were 100%, 100%, 100%, 0, and 100%, respectively. Conclusions: SLN in early-staged cervical cancer patients were mainly distributed in the obturator and external iliac space, pathalogical ultra-staging of SLN could improve the prediction of LN metastasis. Intraoperative SLN mapping is safe, feasible and could predict the state of retroperitoneal LN metastasis in early-staged cervical cancer. SLNB may replace systemic pelvic lymphadenectomy.

[Hemophagocytic lymphohistiocytosis caused by hematogenous disseminated pulmonary tuberculosis: A case report].

Hemophagocytic lymphohistiocytosis (HLH) was a life-threatening syndrome due to the uncontrolled immune activation of cytotoxic T lymphocytes, natural killer (NK) cells, and macrophages. HLH is characterized by primary and secondary causes, the early diagnosis and treatment of patients are closely related to the prognosis and clinical outcome of patients. The clinical presentation is variable but mostly includes prolonged fever, splenomegaly, coagulopathy, hypertriglyceridemia, and hemophagocytosis, none of them is specific and particular for HLH. Tuberculosis (TB) infection is one of the causes of HLH. HLH caused by TB is very rare clinically, but it has a high mortality. For patients with fever of unknown origin, HLH-related clinical manifestations sometimes present before the final diagnosis of TB, and HLH is associated with the most significant mortality rate. This article is mainly about a 28-year-old patient with HLH who suffered from severe TB infection. The patient attended a hospital with a history of 2 months of prolonged fever, 10 days booger and subcutaneous hemorrhage in lower limbs. Before this, he was in good health and denied any history of tuberculosis exposure. Combined with relevant laboratory test results (such as splenomegaly, hemoglobin, platelet count, and hypertriglyceridemia) and clinical manifestations (e.g. fever), the patient was diagnosed with hemophagocytic lymphohistiocytosis, but the etiology of HLH remained to be determined. To confirm the etiology, the patient was asked about the relevant medical history (intermittent low back pain) and was performed chest CT scan, bone marrow biopsy, and fundus photography. Finally, he was diagnosed with hemophagocytic lymphohistiocytosis caused by hematogenous disseminated pulmonary tuberculosis. In response to this, intravenous methylprednisolone and anti-tuberculosis treatment (isoniazid, pyrazinamide, moxifloxacin, and amikacin) were administered to the patient. After more than a month of treatment, the patient recovered from HLH caused by severe TB infection. Therefore, this case suggests that we should be vigilant to the patient who admitted to the hospital with fever for unknown reasons, to diagnose HLH as early as possible and clarify its cause, then perform interventions and treatment, especially HLH secondary to tuberculosis. Also, cases of atypical TB and severe TB should be carefully monitored to achieve early diagnosis and early intervention.

[Research advances on exosomes derived from adipose-derived mesenchymal stem cells in promoting diabetic wound healing].

Impaired healing of diabetic wounds is mainly attributed to its pathological mechanism, and refractory diabetic wounds bring heavy burdens to patients and society. Exosomes derived from stem cells possess the similar ability as stem cells in promoting tissue regeneration and more clinical advantages and are gradually playing important roles in wound healing. In recent years, researches have shown that exosomes derived from adipose-derived mesenchymal stem cells (ADSC-EXOs) can promote the healing of diabetic wounds by participating in various processes of wound healing. This article reviews the pathological mechanism leading to impaired healing of diabetic wounds, the related mechanism and the application prospect of ADSC-EXOs in promoting diabetic wound healing.

RUNX2 Regulates Osteoblast Differentiation via the BMP4 Signaling Pathway.

RUNX2 is a master osteogenic transcription factor, and mutations in RUNX2 cause the inherited skeletal disorder cleidocranial dysplasia (CCD). Studies have revealed that RUNX2 is not only a downstream target of the bone morphogenetic protein (BMP) pathway but can also regulate the expression of BMPs. However, the underlying mechanism of the regulation of BMPs by RUNX2 remains unknown. In this project, we diagnosed a CCD patient with a 7.86-Mb heterozygous deletion on chromosome 6 containing all exons of RUNX2 by multiplex ligation-dependent probe amplification (MLPA) and whole-genome sequencing (WGS). Bone marrow mesenchymal stem cells (BMSCs) were further extracted from patient alveolar bone fragments (CCD-BMSCs), an excellent natural model to explore the possible mechanism. The osteogenic differentiation ability of CCD-BMSCs was severely affected by RUNX2 heterozygous deletion. Also, BMP4 decreased most in BMP ligands, and CHRDL1, a BMP antagonist, was abnormally elevated in CCD-BMSCs. Furthermore, BMP4 treatment essentially rescued the osteogenic capacity of CCD-BMSCs, and RUNX2 overexpression reversed the abnormal expression of BMP4 and CHRDL1. Notably, we constructed CRISPR/Cas9 Runx2+/m MC3T3-E1 cells, which simulated a variant in CCD-BMSCs, to exclude the interference of other gene deletions and the heterogeneity of the genetic background of primary cells, and verified all findings from the CCD-BMSCs. Moreover, the luciferase reporter experiment showed that RUNX2 could inhibit the transcription of CHRDL1. Through immunofluorescence, the inhibitory effect of CHRDL1 on BMP4/Smad signaling was confirmed in MC3T3-E1 cells. These results revealed that RUNX2 regulated the BMP4 pathway by inhibiting CHRDL1 transcription. We collectively identified a novel RUNX2/CHRDL1/BMP4 axis to regulate osteogenic differentiation and noted that BMP4 might be a valuable therapeutic option for treating bone diseases.

[Application of near infrared fluorescence imaging in detection of residual cancer in oral squamous cell carcinoma].

Objective: Local recurrence is the main cause of treatment failure in patients with oral squamous cell carcinoma (OSCC). This study was proposed to investigate the feasibility of near infrared fluorescence (NIF) via indocyanine green (ICG) for monitoring surgical marginal in operation for OSCC patients. Methods: In 35 patients with OSCC treated surgically in the Department of Oral and Maxillofacial Surgery, Nanjing University School of Medicine, from January 2019 to June 2020, ICG (0.75 mg/kg) was administered intravenously via elbow vein at (12±1) hours before surgery, and NIF was performed intraoperatively on the surgical field and the cut edge of the surgically excised specimen, and fluorescence intensity was measured for OSCC tissue and normal oral mucosa, abnormal fluorescence signals were taken and subjected to rapid cryopathological examination. Correlation between NIF tumor boundary grading and pathological tumor boundary grading was analyzed by Spearman correlation analysis. Results: Clear ICG NIF was obtained for tumor lesions in all 35 patients, with a positive rate of 100%. The fluorescence intensity of OSCC tissue was (412.73±146.56) au, which was higher than that of normal oral mucosa tissue [(279.38±82.56) au, P<0.01]. Abnormal fluorescence signals were detected at the tumor bed and the cut edge of the surgical resection specimen in 4 patients, of which 2 cases were pathologically confirmed as cancer cell residue and 2 cases as inflammatory cell infiltration. The rate of positive detection of cut margins using ICG NIF technique in OSCC was 5.7% (2/35). Twenty of the 35 OSCC patients had grade 1, 11 of grade 2, and 4 of grade 3 tumor borders revealed by NIF of surgical resection specimens, which was positively correlated with pathological tumor border (r=0.809, P<0.001). Conclusions: ICG NIF technique can effectively detect the residual cancer cells at the incision margin, which is of great clinical value in reducing local recurrence of OSCC after surgery due to intraoperative cancer residue.

[The prevalence and associated factors of metabolic syndrome among Tibetan pastoralists in transition from nomadic to settled urban environment].

Objective: To study the prevalence and associated factors of metabolic syndrome (MS) among Tibetan pastoralists in transition from high altitude nomadic to settled urbanized environment, especially dietary factors. Methods: The community-based cross-sectional study included 920 Tibetan adults (men 419, women 501). Data were collected using questionnaires, anthropometric measurements, and biomarker tests. Questionnaires included socio-economic, lifestyle characteristics and food consumption. Principal component analysis was used to identify dietary patterns. The risk factors of MS and its components were analyzed by logistic regression model. Results: The prevalence rates of MS and its components were 32.8% (MS), 83.7% (decreased HDL-C), 62.1% (central obesity), 36.7% (elevated blood pressure), 11.8% (elevated TG), and 7.9% (elevated blood glucose), respectively. The prevalence of overweight was 31.2%, obesity 30.3%. Multivariate analysis showed smoking was associated factor for both of decreased HDL-C (OR=1.239, 95%CI: 1.025-1.496) and elevated TG (OR=1.277, 95%CI: 1.038-1.571). Alcohol drinking appeared as associated factor of elevated TG (OR=1.426, 95%CI: 1.055-1.927). However, physical activity showed as a protective factor for central obesity, decreased HDL-C, and elevated TG. With the increase of age, the adherence to the urban and western dietary patterns decreased, and that to the pastoral dietary pattern increased. By quintiles of dietary pattern scores, the urban dietary pattern was significantly associated with MS (trend test P=0.016). Conclusions: Tibetan pastoralists had high prevalence of both MS and obesity. Smoking, alcohol drinking, the transition from pastoral dietary pattern to urban dietary pattern and inadequate physical activity served as associated factors for MS and its components.

[The safety and efficacy of anlotinib in combination with stereotactic radiotherapy for the treatment of brain metastases from non-small cell lung cancer].

Objective: To investigate the efficacy and safety of anti-vascular drug anlotinib in combination with stereotactic radiosurgery (SRS) for the treatment of brain metastases from non-small cell lung cancer (NSCLC). Methods: A total of 46 patients diagnosed with NSCLC brain metastases and treated with SRS in Peking University Third Hospital were included from October 2017 to June 2019. Of these, 21 patients (33 lesions) received anlotinib combined with SRS (combined treatment group), while 25 patients (35 lesions) only received SRS (SRS-alone group). The data of combined treatment group and SRS-alone group were compared, including remission rate of intracranial hypertension, response rate (RR) of local control of intracranial lesions, incidence of radiation-induced brain necrosis and intracranial progression-free survival (iPFS). The medication and adverse reactions of anlotinib in the combined treatment group were recorded. Results: The remission rate of intracranial hypertension in the combined treatment group was 71.4% (15/21), which was significantly higher than that in the SRS-alone group [12.0% (3/25), P<0.001). However, the RR of combined treatment group and SRS-alone group was 80.9% (17/21) and 60.0% (15/25), respectively, with no statistically significant difference (P=0.289). The incidence of radiation-induced brain necrosis in the combined treatment group was 3.0% (1/33), which was significantly lower than that in the SRS-alone group [20.0% (7/35), P=0.030]. The iPFS of the combination treatment group was (13.9±2.4) months, which was significantly longer than that in the SRS-alone group [(11.4±1.8) months, P<0.001]. The medication time of anlotinib in the combined treatment group was 6 (6, 18) weeks. One patient developed grade Ⅰ hypertension and one patient developed grade Ⅰ hand-foot syndrome. The incidence of drug-related adverse reactions was 9.5% (2/21). Conclusions: Anlotinib in combination with SRS may relieve brain edema, reduce the rate of radiation-induced brain necrosis, and the rate of drug-related adverse reactions were low.

Identification of Malus halliana R2R3-MYB gene family under iron deficiency stress and functional characteristics of MhR2R3-MYB4 in Arabidopsis thaliana.

Iron (Fe) is an essential element for plant growth and development. Fe deficiency can trigger leaf chlorosis and reduce fruit yield. Therefore, it is necessary to explore transcription factors in response to Fe deficiency stress. A total of 29 MhR2R3-MYB transcription factors were identified based on the transcriptome of Malus halliana under Fe deficiency stress. A comprehensive analysis of physical and chemical properties, gene structures, conserved motif composition, evolutionary relationship and chromosome distribution was performed. Subsequently, based on the transcriptome, 14 genes with the most significant expression under Fe deficiency stress were screened for qRT-PCR verification. Among them,the functional characteristics of MhR2R3-MYB4 (MD05G1089600) were further studied in Arabidopsis thaliana. Expression of 13 out of these 14 genes was upregulated, only one was downregulated. Maximum upregulation of MhR2R3-MYB4 under Fe deficiency was 36.39-fold and 58.21-fold compared with day 0 in leaves and roots, respectively. Overexpression of MhR2R3-MYB4 enhanced tolerance to Fe deficiency in A. thaliana and led to multiple biochemical changes: transgenic lines have higher chl a, chl b and Fe2+ content, higher enzyme activity (SOD, POD, CAT and FCR) and lower chlorosis than the wild type in Fe deficiency conditions. We suggest that MhR2R3-MYB4 plays an important part in Fe deficiency stress, which may contribute to improve Fe deficiency tolerance of apple in future.

[The peripheral blood inflammatory markers in Ménière's disease patients with and without migraine].

Objective: To investigate the peripheral blood inflammatory markers including white blood cell count (WBC), monocytes, neutrophils, lymphocytes, platelets, neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), high-density lipoprotein(HDL-C), low-density lipoprotein and fibrinogen (FIB) in Ménière's disease (MD) patients with and without migraine, and to explore the relationship between the inflammatory response with MD and migraine. Methods: The general physical condition, clinical manifestations, pure-tone audiometry, and peripheral blood inflammatory markers of 92 unilateral MD patients who were hospitalized in Peking University People's Hospital for surgical treatment from January 2017 to January 2021 were continuously collected. Meanwhile, 50 healthy controls matched with age and sex were included, and their general physical conditions and peripheral blood inflammatory markers were also collected. This study consisted of two parts. First, the differences in epidemical characteristics and peripheral blood inflammatory markers between MD patients and healthy controls were compared by univariate analysis. Second, all 92 MD patients were divided into two subgroups according to whether they were accompanied by migraine. The clinical characteristics and peripheral blood inflammatory markers of MD patients with and without migraine were compared by univariate analysis. Thereafter, binary Logistic regression was used to analyze the related factors of whether MD patients were accompanied with migraine. Results: Compared with the healthy control group, the peripheral blood WBC, neutrophils and FIB of MD patients were significantly increased (all P<0.05). Compared with MD patients without migraine, MD patients with migraine had higher female prevalence, longer disease history, lower low-frequency hearing threshold, higher frequency of vertigo attacks and higher HDL-C levels (all P<0.05), meanwhile, female, frequency of vestibular attacks and HDL-C were independent related factors of whether MD patients were accompanied with migraine. Conclusion: The occurrence of MD and migraine may be related to the inflammatory response. The level of anti-inflammatory factors in the blood of MD patients with migraine are higher, suggesting that the inflammatory response status of MD patients with and without migraine is different.