Mycophenolic acid (MPA), the active moiety of both mycophenolate mofetil (MMF) and enteric-coated mycophenolate sodium (EC-MPS), serves as a primary immunosuppressant for maintaining solid organ transplants. Therapeutic drug monitoring (TDM) enhances treatment outcomes through tailored approaches. This study aimed to develop an evidence-based guideline for MPA TDM, facilitating its rational application in clinical settings. The guideline plan was drawn from the Institute of Medicine and World Health Organization (WHO) guidelines. Using the Delphi method, clinical questions and outcome indicators were generated. Systematic reviews, Grading of Recommendations Assessment, Development, and Evaluation (GRADE) evidence quality evaluations, expert opinions, and patient values guided evidence-based suggestions for the guideline. External reviews further refined the recommendations. The guideline for the TDM of MPA (IPGRP-2020CN099) consists of four sections and 16 recommendations encompassing target populations, monitoring strategies, dosage regimens, and influencing factors. High-risk populations, timing of TDM, area under the curve (AUC) versus trough concentration (C₀), target concentration ranges, monitoring frequency, and analytical methods are addressed. Formulation-specific recommendations, initial dosage regimens, populations with unique considerations, pharmacokinetic-informed dosing, body weight factors, pharmacogenetics, and drug‍-‍drug interactions are covered. The evidence-based guideline offers a comprehensive recommendation for solid organ transplant recipients undergoing MPA therapy, promoting standardization of MPA TDM, and enhancing treatment efficacy and safety.
Mycophenolic Acid/administration & dosage* ; Drug Monitoring/methods* ; Humans ; Organ Transplantation ; Immunosuppressive Agents/administration & dosage* ; Delphi Technique

Objective:To investigate the association between cognitive impairment and manifestation of interictal electroencephalogram (EEG) in epilepsy patients, and the early assessment significance of the Grand Total Electroencephalography (GTE) score.Methods:A totall of 100 patients with primary epilepsy admitted to the Department of Neurology of the Affiliated Hospital of Yangzhou University were continuously collected from January 2019 to January 2024, and they were classified according to the latest version of the epilepsy classification by the International League Against Epilepsy in 2017. General information of all research subjects was recorded, including age, gender, educational level, etc. The disease details of epilepsy patients were recorded, including seizure duration, severity, seizure precursors, post seizure status, and use of anti-seizure medications (ASM). The survey scales and questionnaires used included the interictal GTE, Montreal Cognitive Assessment (MoCA), Hamilton Anxiety Scale, Hamilton Depression Scale, National Hospital Seizure Severity Scale (NHS3), Status Epilepticus Severity Score (STESS). All research subjects were classified into normal cognitive (NC) group, mild cognitive impairment (MCI) group, and dementia group according to MoCA score. Comparisons among multiple groups and pairwise comparisons were conducted. The correlation between 2 variables was analyzed using Spearman rank correlation analysis, and multiple linear regression analysis was employed to screen variables that have an impact on cognitive impairment. The receiver operating characteristic curve was plotted to determine the optimal cut-off point for predicting cognitive impairment in epilepsy.Results:According to the MoCA score, there were 32 (32%) patients in the NC group, 49 (49%) patients in the MCI group, and 19 (19%) patients in the dementia group. There were statistically significant differences in age [(26.31±10.01) years, (43.96±16.19) years, (57.68±16.83) years,respectively; F=29.440, P<0.001], education ( χ2=28.894, P<0.001), ASM ( χ2=11.258, P<0.017), STESS score [2.00(1.75, 2.25), 2.00(2.00, 3.00), 3.50(2.75, 4.25),respectively; H=12.646, P=0.002], STESS score>2 ( χ2=10.075, P=0.006), frequency of rhythmic background activity ( H=17.429, P<0.001), diffuse slow activity ( H=42.033, P<0.001), reactivity of the rhythmic background activity ( H=15.206, P<0.001), paroxysmal activity ( H=25.279, P<0.001), sharp wave activity ( H=15.492, P<0.001) and total GTE score [1 (1, 3), 6 (2, 8), 8 (7, 11),respectively; H=47.871, P<0.001] among the 3 groups. A significant negative correlation was observed between cognitive level (MoCA scores) and total GTE score ( ρ=-0.766, P<0.001), frequency of rhythmic background activity ( ρ=-0.520, P<0.001), diffuse slow activity ( ρ=-0.734, P<0.001), reactivity of the rhythmic background activity ( ρ=-0.438, P<0.001), paroxysmal activity ( ρ=-0.566, P<0.001), and sharp wave activity ( ρ=-0.407, P<0.001). The results of multiple linear regression analysis indicated that total GTE score ( t=-5.566, P<0.001), diffuse slow activity ( t=-2.548, P=0.014), reactivity of the rhythmic background activity ( t=-3.891, P<0.001), paroxysmal activity ( t=-3.139, P=0.003), age ( t=-5.493, P<0.001), education ( t=3.379, P=0.001), and STESS ( t=-2.183, P=0.033) were independent risk factors for cognitive impairment. In evaluating the cognitive impairment of epilepsy patients, the GTE score had a certain sensitivity (75.0%) and specificity (93.8%), with an optimal critical point value of 5. Conclusions:The interictal EEG of patients with poorer cognitive function is mainly characterized by an increase in slow waves and a decrease in overall background. The increase in slow waves, poor background responsiveness, paroxysmal activity and a high total GTE score may be important factors in predicting the outcome of cognitive impairment in epilepsy.

Objective:To analyze postictal electroencephalographic(EEG) characteristics of patients with status epilepticus (SE) based on the score of grand total electroencephalography (GTE), and explore the relationship between electroencephalographic characteristics of SE and clinical prognosis.Methods:A total of 110 SE patients were enrolled in the Department of Neurology, the Affiliated Hospital of Yangzhou University from September 1, 2021 to September 1, 2023. EEG and GTE scores were performed in all patients after seizures (0-2 days after the cessation of SE). After one year of discharge, the medication and seizure status of patients were followed up by phone or outpatient visits. The seizure outcomes were recorded according to the international league against epilepsy (ILAE) seizure outcome classification, with favorable outcomes defined as good prognosis group ( n=54) and unfavorable outcomes defined as poor prognosis group ( n=56). SPSS 27.0 software was used for statistical analysis. Binary Logistic regression analysis was employed to screen impact variables of prognosis. The receiver operating characteristic(ROC) curve was plotted to determine the optimal cut-off point for predicting prognosis in epilepsy. Results:There were statistically significant differences in the total GTE score(2(1, 4), 8(5, 10); Z=-6.837, P<0.001), diffuse slow activity(0(0, 1), 2(0, 2); Z=-6.495, P<0.001), reactivity of the rhythmic background activity(0(0, 0), 0(0, 1); Z=-2.705, P=0.007), paroxysmal activity(0(0, 0), 1.5(0, 3.0); Z=-4.420, P<0.001), focal disturbances(0(0, 0), 0(0, 0); Z=-2.130, P=0.033), and sharp wave activity(0(0, 2), 2(2, 3); Z=-5.714, P<0.001)between the good prognosis group and poor prognosis group. The differences in EEG results among SE patients with different types of epileptic seizures were statistically significant in terms of frequency of rhythmic background activity, diffuse slow activity, reactivity of rhythmic background activity and total GTE score (all P<0.05). The differences in EEG results between SE patients with clear and unknown causes were statistically significant in terms of paroxysmal activity and focal disturbances(both P<0.05). The results of binary Logistic regression analysis showed that independent factors associated with the prognosis of SE included medication adherence ( B=-0.496, OR=0.609, 95% CI=0.395-0.940, P=0.025), diffuse slow activity( B=1.580, OR=4.854, 95% CI=1.586-14.855, P=0.006), sharp wave activity( B=0.824, OR=2.280, 95% CI=1.210-4.297, P=0.011), and total GTE score ( B=0.561, OR=1.753, 95% CI=1.360-2.259, P<0.001). In evaluating the prognosis of SE, the GTE score had a certain sensitivity (74.6%) and specificity (85.1%), with a optimal cut-off point of 6. Conclusions:The differences in EEG results among SE patients with different types of epileptic seizures were statistically significant in terms of frequency of rhythmic background activity, diffuse slow activity, reactivity of rhythmic background activity. The appearance of diffuse slow activity and sharp wave activity in the electroencephalogram of SE patients indicates poor prognosis, and the total GTE score≥6 may be a strong predictor of poor prognosis. However, good medication adherence is a protective factor for epilepsy recurrence.

Objective:To evaluate the clinical efficacy of the modified medial gastrocnemius myocutaneous flap (MGMF) with extended anterior, posterior and (or) inferior boundaries.Methods:From January 2002 to September 2022, modified MGMFs were applied onto 33 patients who received reconstructive surgery for soft-tissue defects around knee or in calf, in the Department of Orthopaedics, the Second Xiangya Hospital of Central South University. The size of defects ranged from 10 cm×4 cm to 22 cm×12 cm, and the flap size ranged from 15 cm×6 cm to 28 cm×14 cm. Twenty-five patients had the complication of chronic osteomyelitis. The boundaries of a modified MGMF were as follows: the anterior boundary was the anterior border of the tibia, where the posterior boundary at 3.0 cm lateral to the posterior midline, the proximal boundary at the popliteal fossa crease, and the distal boundary at the plane 2.0 cm above the tip of medial malleolus. The anterior edge of the modified MGMF was designed running along the medial edge of the defect and its curved extension line. Pretibial skin was equally divided into 9 zones, with the 1st to 9th zones from proximal to distal in sequence. Postoperative routine anti-infection treatment was offered. All patients were included in the postoperative follow-up through outpatient visits, telephone or WeChat interviews. Flap viability and wound healing in both donor and recipient sites were evaluated. Function of the affected limb was assessed using the evaluation criteria established by Punor et al.Results:All patients were included in the follow-up for 1 to 169 (median duration: 9)months. The 33 modified MGMFs included MGMFs with extended boundary of anterior ( n=18), inferior ( n=5), anterior combined with inferior ( n=6), posterior combined with anterior ( n=2), and posterior combined with inferior ( n=2) boundaries. Twenty-nine (87.9%) flaps survived completely. Partial necrosis occurred in 4 flaps(12.1%)(2 flaps with extended anterior boundary and 2 flaps with extended inferior boundary). The anterior margins of 26 flaps (78.7%) with extended anterior boundary alone or in combination with extended inferior or posterior boundary exceeded the medial edge of the tibia by 1.0-4.5 (mean, 2.1) cm, and 3 of them reached the anterior edge of tibia. Fourteen (42.4%) modified MGMFs were used to reconstruct the defects involving 1/3 of distal calf, and the distal ends of these defects were located in the 7th ( n=8) or 8th ( n= 6) zone. All the skin grafts in the donor sites survived. During follow-up, 31 patients (93.9%) showed no sign of infection, and 2 patients (6.1%) who had recurrence of chronic osteomyelitis. Functions of the affected limbs were excellent ( n=25), good ( n=6) and fair ( n=2) by Punor et al. Conclusion:Modified MGMF with extended anterior, posterior and (or) inferior boundaries is clinically feasible. It offers advantages of easier design and operation. It can be used to reconstruct a more distal, wider and larger defect as well as broadens the application of the MGMF.

Objective:To investigate the clinicopathological characteristics, immunophenotype and differential diagnosis of atypical forms of microglandular hyperplasia of the cervix (AMGH).Methods:A total of 29 cases of AMGH diagnosed at the Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China from January 2010 to December 2024 were analyzed. Relevant clinical and pathological data of the patients were collected using the electronic medical record system and medical records copied from the outside hospitals. The patients were followed up.Results:Among the 29 cases, 28 were consultation cases, 22 (79%) of the 28 cases were considered as glandular neoplastic lesions by the original institutions. The nature of the lesion was uncertain in 1 case, the diagnosis was suspicious for AMGH in another 1 case, and only 4 cases were clearly diagnosed as AMGH. The median age of the 29 patients was 44 (43, 48) years. Eighteen (62%) of the 29 cases presented as cervical polyp. Twelve of the 16 tested cases were negative for human papillomavirus. The pathological presentation was complex and diverse, including solid, trabecular, cribriform, and papillary patterns, forming pseudo-invasive structures. The glandular epithelium and proliferating reserve cells had diverse morphologies, which presented with abundant eosinophilic cytoplasm or clear cytoplasm. Signet-ring or hobnail cells were also seen. The nuclear atypia was mild, with 0-7 mitotic figures per 10 HPF. Immature squamous metaplasia was noted. The stroma showed edema, myxoid change and hyaline degeneration, accompanied by infiltration of acute and chronic inflammatory cells. Immunohistochemistry demonstrated that p16 was negative in 8/16 of the cases or patchy positive in the other 8/16, Ki-67 positive rate was less than 10% in all 16 cases, p53 was wild phenotype (9/9), and carcinoembryonic antigen was negative in 4/5 cases and focally positive in 1/5 cases, while p63 was positive in 6/9 of the tested cases.Conclusions:AMGH is a benign non-neoplastic lesion of the cervical glands. Half of the cases occur in perimenopausal or postmenopausal women, often presenting as polypoid hyperplasia or localized cervical thickening/elevation with a friable, fragile texture. Microscopically, it may show a pseudoinvasive pattern, making it prone to misdiagnosis as a malignant lesion. Thus, differentiation from cervical adenocarcinoma, clear cell carcinoma and microglandular endometrioid carcinoma is required. Integration of clinical history, immunohistochemistry and molecular testing may aid in the differential diagnosis.

Objective:To analyze postictal electroencephalographic(EEG) characteristics of patients with status epilepticus (SE) based on the score of grand total electroencephalography (GTE), and explore the relationship between electroencephalographic characteristics of SE and clinical prognosis.Methods:A total of 110 SE patients were enrolled in the Department of Neurology, the Affiliated Hospital of Yangzhou University from September 1, 2021 to September 1, 2023. EEG and GTE scores were performed in all patients after seizures (0-2 days after the cessation of SE). After one year of discharge, the medication and seizure status of patients were followed up by phone or outpatient visits. The seizure outcomes were recorded according to the international league against epilepsy (ILAE) seizure outcome classification, with favorable outcomes defined as good prognosis group ( n=54) and unfavorable outcomes defined as poor prognosis group ( n=56). SPSS 27.0 software was used for statistical analysis. Binary Logistic regression analysis was employed to screen impact variables of prognosis. The receiver operating characteristic(ROC) curve was plotted to determine the optimal cut-off point for predicting prognosis in epilepsy. Results:There were statistically significant differences in the total GTE score(2(1, 4), 8(5, 10); Z=-6.837, P<0.001), diffuse slow activity(0(0, 1), 2(0, 2); Z=-6.495, P<0.001), reactivity of the rhythmic background activity(0(0, 0), 0(0, 1); Z=-2.705, P=0.007), paroxysmal activity(0(0, 0), 1.5(0, 3.0); Z=-4.420, P<0.001), focal disturbances(0(0, 0), 0(0, 0); Z=-2.130, P=0.033), and sharp wave activity(0(0, 2), 2(2, 3); Z=-5.714, P<0.001)between the good prognosis group and poor prognosis group. The differences in EEG results among SE patients with different types of epileptic seizures were statistically significant in terms of frequency of rhythmic background activity, diffuse slow activity, reactivity of rhythmic background activity and total GTE score (all P<0.05). The differences in EEG results between SE patients with clear and unknown causes were statistically significant in terms of paroxysmal activity and focal disturbances(both P<0.05). The results of binary Logistic regression analysis showed that independent factors associated with the prognosis of SE included medication adherence ( B=-0.496, OR=0.609, 95% CI=0.395-0.940, P=0.025), diffuse slow activity( B=1.580, OR=4.854, 95% CI=1.586-14.855, P=0.006), sharp wave activity( B=0.824, OR=2.280, 95% CI=1.210-4.297, P=0.011), and total GTE score ( B=0.561, OR=1.753, 95% CI=1.360-2.259, P<0.001). In evaluating the prognosis of SE, the GTE score had a certain sensitivity (74.6%) and specificity (85.1%), with a optimal cut-off point of 6. Conclusions:The differences in EEG results among SE patients with different types of epileptic seizures were statistically significant in terms of frequency of rhythmic background activity, diffuse slow activity, reactivity of rhythmic background activity. The appearance of diffuse slow activity and sharp wave activity in the electroencephalogram of SE patients indicates poor prognosis, and the total GTE score≥6 may be a strong predictor of poor prognosis. However, good medication adherence is a protective factor for epilepsy recurrence.

Objective:To evaluate the clinical efficacy of the modified medial gastrocnemius myocutaneous flap (MGMF) with extended anterior, posterior and (or) inferior boundaries.Methods:From January 2002 to September 2022, modified MGMFs were applied onto 33 patients who received reconstructive surgery for soft-tissue defects around knee or in calf, in the Department of Orthopaedics, the Second Xiangya Hospital of Central South University. The size of defects ranged from 10 cm×4 cm to 22 cm×12 cm, and the flap size ranged from 15 cm×6 cm to 28 cm×14 cm. Twenty-five patients had the complication of chronic osteomyelitis. The boundaries of a modified MGMF were as follows: the anterior boundary was the anterior border of the tibia, where the posterior boundary at 3.0 cm lateral to the posterior midline, the proximal boundary at the popliteal fossa crease, and the distal boundary at the plane 2.0 cm above the tip of medial malleolus. The anterior edge of the modified MGMF was designed running along the medial edge of the defect and its curved extension line. Pretibial skin was equally divided into 9 zones, with the 1st to 9th zones from proximal to distal in sequence. Postoperative routine anti-infection treatment was offered. All patients were included in the postoperative follow-up through outpatient visits, telephone or WeChat interviews. Flap viability and wound healing in both donor and recipient sites were evaluated. Function of the affected limb was assessed using the evaluation criteria established by Punor et al.Results:All patients were included in the follow-up for 1 to 169 (median duration: 9)months. The 33 modified MGMFs included MGMFs with extended boundary of anterior ( n=18), inferior ( n=5), anterior combined with inferior ( n=6), posterior combined with anterior ( n=2), and posterior combined with inferior ( n=2) boundaries. Twenty-nine (87.9%) flaps survived completely. Partial necrosis occurred in 4 flaps(12.1%)(2 flaps with extended anterior boundary and 2 flaps with extended inferior boundary). The anterior margins of 26 flaps (78.7%) with extended anterior boundary alone or in combination with extended inferior or posterior boundary exceeded the medial edge of the tibia by 1.0-4.5 (mean, 2.1) cm, and 3 of them reached the anterior edge of tibia. Fourteen (42.4%) modified MGMFs were used to reconstruct the defects involving 1/3 of distal calf, and the distal ends of these defects were located in the 7th ( n=8) or 8th ( n= 6) zone. All the skin grafts in the donor sites survived. During follow-up, 31 patients (93.9%) showed no sign of infection, and 2 patients (6.1%) who had recurrence of chronic osteomyelitis. Functions of the affected limbs were excellent ( n=25), good ( n=6) and fair ( n=2) by Punor et al. Conclusion:Modified MGMF with extended anterior, posterior and (or) inferior boundaries is clinically feasible. It offers advantages of easier design and operation. It can be used to reconstruct a more distal, wider and larger defect as well as broadens the application of the MGMF.

Objective:To investigate the association between cognitive impairment and manifestation of interictal electroencephalogram (EEG) in epilepsy patients, and the early assessment significance of the Grand Total Electroencephalography (GTE) score.Methods:A totall of 100 patients with primary epilepsy admitted to the Department of Neurology of the Affiliated Hospital of Yangzhou University were continuously collected from January 2019 to January 2024, and they were classified according to the latest version of the epilepsy classification by the International League Against Epilepsy in 2017. General information of all research subjects was recorded, including age, gender, educational level, etc. The disease details of epilepsy patients were recorded, including seizure duration, severity, seizure precursors, post seizure status, and use of anti-seizure medications (ASM). The survey scales and questionnaires used included the interictal GTE, Montreal Cognitive Assessment (MoCA), Hamilton Anxiety Scale, Hamilton Depression Scale, National Hospital Seizure Severity Scale (NHS3), Status Epilepticus Severity Score (STESS). All research subjects were classified into normal cognitive (NC) group, mild cognitive impairment (MCI) group, and dementia group according to MoCA score. Comparisons among multiple groups and pairwise comparisons were conducted. The correlation between 2 variables was analyzed using Spearman rank correlation analysis, and multiple linear regression analysis was employed to screen variables that have an impact on cognitive impairment. The receiver operating characteristic curve was plotted to determine the optimal cut-off point for predicting cognitive impairment in epilepsy.Results:According to the MoCA score, there were 32 (32%) patients in the NC group, 49 (49%) patients in the MCI group, and 19 (19%) patients in the dementia group. There were statistically significant differences in age [(26.31±10.01) years, (43.96±16.19) years, (57.68±16.83) years,respectively; F=29.440, P<0.001], education ( χ2=28.894, P<0.001), ASM ( χ2=11.258, P<0.017), STESS score [2.00(1.75, 2.25), 2.00(2.00, 3.00), 3.50(2.75, 4.25),respectively; H=12.646, P=0.002], STESS score>2 ( χ2=10.075, P=0.006), frequency of rhythmic background activity ( H=17.429, P<0.001), diffuse slow activity ( H=42.033, P<0.001), reactivity of the rhythmic background activity ( H=15.206, P<0.001), paroxysmal activity ( H=25.279, P<0.001), sharp wave activity ( H=15.492, P<0.001) and total GTE score [1 (1, 3), 6 (2, 8), 8 (7, 11),respectively; H=47.871, P<0.001] among the 3 groups. A significant negative correlation was observed between cognitive level (MoCA scores) and total GTE score ( ρ=-0.766, P<0.001), frequency of rhythmic background activity ( ρ=-0.520, P<0.001), diffuse slow activity ( ρ=-0.734, P<0.001), reactivity of the rhythmic background activity ( ρ=-0.438, P<0.001), paroxysmal activity ( ρ=-0.566, P<0.001), and sharp wave activity ( ρ=-0.407, P<0.001). The results of multiple linear regression analysis indicated that total GTE score ( t=-5.566, P<0.001), diffuse slow activity ( t=-2.548, P=0.014), reactivity of the rhythmic background activity ( t=-3.891, P<0.001), paroxysmal activity ( t=-3.139, P=0.003), age ( t=-5.493, P<0.001), education ( t=3.379, P=0.001), and STESS ( t=-2.183, P=0.033) were independent risk factors for cognitive impairment. In evaluating the cognitive impairment of epilepsy patients, the GTE score had a certain sensitivity (75.0%) and specificity (93.8%), with an optimal critical point value of 5. Conclusions:The interictal EEG of patients with poorer cognitive function is mainly characterized by an increase in slow waves and a decrease in overall background. The increase in slow waves, poor background responsiveness, paroxysmal activity and a high total GTE score may be important factors in predicting the outcome of cognitive impairment in epilepsy.

Objective:To investigate the clinicopathological characteristics, immunophenotype and differential diagnosis of atypical forms of microglandular hyperplasia of the cervix (AMGH).Methods:A total of 29 cases of AMGH diagnosed at the Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China from January 2010 to December 2024 were analyzed. Relevant clinical and pathological data of the patients were collected using the electronic medical record system and medical records copied from the outside hospitals. The patients were followed up.Results:Among the 29 cases, 28 were consultation cases, 22 (79%) of the 28 cases were considered as glandular neoplastic lesions by the original institutions. The nature of the lesion was uncertain in 1 case, the diagnosis was suspicious for AMGH in another 1 case, and only 4 cases were clearly diagnosed as AMGH. The median age of the 29 patients was 44 (43, 48) years. Eighteen (62%) of the 29 cases presented as cervical polyp. Twelve of the 16 tested cases were negative for human papillomavirus. The pathological presentation was complex and diverse, including solid, trabecular, cribriform, and papillary patterns, forming pseudo-invasive structures. The glandular epithelium and proliferating reserve cells had diverse morphologies, which presented with abundant eosinophilic cytoplasm or clear cytoplasm. Signet-ring or hobnail cells were also seen. The nuclear atypia was mild, with 0-7 mitotic figures per 10 HPF. Immature squamous metaplasia was noted. The stroma showed edema, myxoid change and hyaline degeneration, accompanied by infiltration of acute and chronic inflammatory cells. Immunohistochemistry demonstrated that p16 was negative in 8/16 of the cases or patchy positive in the other 8/16, Ki-67 positive rate was less than 10% in all 16 cases, p53 was wild phenotype (9/9), and carcinoembryonic antigen was negative in 4/5 cases and focally positive in 1/5 cases, while p63 was positive in 6/9 of the tested cases.Conclusions:AMGH is a benign non-neoplastic lesion of the cervical glands. Half of the cases occur in perimenopausal or postmenopausal women, often presenting as polypoid hyperplasia or localized cervical thickening/elevation with a friable, fragile texture. Microscopically, it may show a pseudoinvasive pattern, making it prone to misdiagnosis as a malignant lesion. Thus, differentiation from cervical adenocarcinoma, clear cell carcinoma and microglandular endometrioid carcinoma is required. Integration of clinical history, immunohistochemistry and molecular testing may aid in the differential diagnosis.

Objective:To observe the expression of CD36 in hepatocellular carcinoma tissues and cell lines, and to investigate the effects of CD36 on the proliferation and migration abilities of human hepatocellular carcinoma cell lines and human hepatocellular carcinoma cell xenograft models in nude mice.Methods:Differences in the expression levels of CD36 transcripts in 371 hepatocellular carcinoma and paracancerous tissues were analyzed based on information from The Cancer Genome Atlas (TCGA) database. Cancer tissues and corresponding paracancerous tissues of 48 hepatocellular carcinoma patients who were diagnosed and underwent surgical treatment at the Affiliated Hospital of Yangzhou University from January 2019 to February 2021 were prospectively collected, and the levels of CD36 mRNA in the tissues were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) method. Western blotting was used to detect CD36 protein levels in human hepatocellular carcinoma cell lines Huh7 and HCCLM3 and human normal liver cell line LO2. Plasmids containing CD36 interfering sequences and empty plasmids were transfected into Huh7 cells or HCCLM3 cells for sh-CD36 group and control group, respectively. The CCK-8 assay was used to detect the proliferation ability (expressed as absorbance value) of cells in each group at 0, 12, 24, 36, 48 and 60 h of culture, and the scratch healing assay and Transwell assay were used to detect the migration ability of cells in each group. The Huh7 cells of sh-CD36 group or control group were injected into the axillary subcutis of BALB/c nude mice, with 4 mice in each group, to construct nude mice models of human hepatocellular carcinoma xenografts; the long and short diameters of tumor were measured weekly after 1 week of inoculation, and the tumor volume was calculated. The nude mice were put to death after 5 weeks of inoculation, and the tumor specimens were collected and weighed; the tumor cell morphology was observed under the microscope, and the expressions of CD36 and Ki-67 proteins in the tumor tissues was detected by immunohistochemistry (IHC).Results:Analysis of the data from the TCGA database showed that the level of CD36 transcripts was higher in hepatocellular carcinoma tissues compared with that in paracancerous tissues (4.2±1.8 vs. 3.2±1.5, t = 2.28, P = 0.035). Tissues detection using qRT-PCR in 48 patients with hepatocellular carcinoma showed that the relative expression of CD36 mRNA in hepatocellular carcinoma tissues was higher than that in paracancerous tissues (0.76±0.26 vs. 0.48±0.23, t = 3.52, P < 0.001). Western blotting assay showed that CD36 protein level in Huh7 and HCCLM3 cells was higher than that in LO2 cells, which were (1.42±0.11) times and (1.68±0.16) times higher than LO2 cells, respectively (both P < 0.001). At the mRNA and protein levels, the CD36 of Huh7 and HCCLM3 cells in the sh-CD36 group was lower than that in the corresponding control group (both P < 0.001). CCK-8 assay showed that the proliferative ability of Huh7 cells and HCCLM3 cells in the sh-CD36 group was lower than that in the corresponding control group after 36 and 24 h of culture (both P < 0.01). Scratch healing assay showed that the scratch healing rates of Huh7 cells [(12±3)% vs. (30±5)%, t = 4.01, P < 0.001] and HCCLM3 cells [(15±4)% vs. (29±5)%, t = 4.16, P < 0.001] in the sh-CD36 group were lower than those in the corresponding control group at 48 h of culture; Transwell assay showed that the number of Huh7 cells [(46±6) cells/field of view vs. (88± 6) cells/field of view, t = 5.56, P < 0.001] and HCCLM3 cells [(42±5) cells/field of view vs. (82±7) cells/field of view, t = 5.34, P < 0.001] penetrating into the membrane in 24 h in the sh-CD36 group was less than that in the corresponding control group. Five weeks after subcutaneous injection, the tumor volume [(682±268) mm 3vs. (1 375±512) mm 3, t = 4.73, P = 0.006] and tumor mass [(432±95) mg/mouse vs. (871±109) mg/mouse, t = 6.57, P < 0.001] of nude mice injected with Huh7 cells of the sh-CD36 group were lower than those of nude mice injected with Huh7 cells of the control group; under the microscope, the density of tumor cells in transplanted tumor specimens of nude mice injected with Huh7 cells of the sh-CD36 group was lower than that in nude mice injected with Huh7 cells of the control group, and the expression levels of both CD36 and Ki-67 proteins were also low. Conclusions:CD36 expression is up-regulated in cancer tissues of hepatocellular carcinoma patients and human hepatocellular carcinoma cell lines Huh7 and HCCLM3, and it may associate with cell proliferation and migration of hepatocellular carcinoma. Knockdown of CD36 expression significantly inhibits the proliferation and migration abilities of hepatocellular carcinoma cells in vitro, and inhibits the tumors of human hepatocellular carcinoma cell xenograft models in nude mice.