ObjectiveTo investigate the mechanism of action of Huangqi Guizhi Wuwutang （HGWT） against cerebral ischemia-reperfusion injury （CIRI） based on bioinformatics and experimental validation. MethodsBiological informatics methods were used to screen for active components of HGWT and their targets. The GEO database was utilized to obtain CIRI-related differentially expressed genes （DEGs）， and platforms such as GeneCards were used to identify disease targets. Venn diagram analysis was conducted to identify overlapping targets， followed by protein-protein interaction （PPI）， gene ontology （GO）， and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis， as well as immune infiltration and immune cell differential analysis. Core genes （Hub genes） were screened using LASSO regression and ROC curves， and molecular docking was used to validate the binding efficiency between the active components of the drug and the core targets. A rat CIRI model was established， with rats randomly divided into five groups （n=10）： Sham surgery group （Sham）， model group （MG）， and low-dose （LD，5.3 g·kg^-1）， medium-dose （MD，10.6 g·kg^-1）， and high-dose （HD，21.2 g·kg^-1） HGWT groups. From 3 days before modeling to 7 days after surgery， oral administration was performed daily： Sham and MG groups received physiological saline， while each drug group received the corresponding dose of HGWT. Hematoxylin-eosin （HE） staining， Nissl staining， and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL staining） were used to assess the repair effects of HGWT on neural damage. Western blot analysis was used to detect B-cell lymphoma-2 protein （Bcl-2）， Bcl-2-associated X protein （Bax）， signal transducer and activator of transcription 3 （STAT3）， phosphorylated STAT3 ［p-STAT3 （Tyr705）］， protein kinase B1 （Akt1）， and phosphorylated Akt1 ［p-Akt1 （Ser473）］， among other target proteins. ResultsAfter screening， 56 common target points of DEGs-disease-drug were obtained. GO and KEGG analyses indicated that HGWT primarily functions in pathways such as apoptosis， oxidative stress， and inflammatory responses. Immune infiltration analysis revealed a significant association between HGWT's anti-CIRI activity and immune cells such as Th17 cells and myeloid-derived suppressor cells （MDSCs） （P0.01）. LASSO-ROC analysis identified Akt1， Caspase-3， glycogen synthase kinase-3β （GSK-3β）， and STAT3 as core genes. Molecular docking confirmed that Hub genes exhibit significant binding affinity with the active components of HGWT （binding energy ≤ -5 kJ·mol^-1）（1 cal≈4.186 J）. Animal experiment results showed that compared with the sham group， the MG group exhibited significant neuronal necrosis， nuclear condensation， and vacuolar degeneration in rat brains， with a significant decrease in Nissl body density （P0.01） and increased neuronal apoptosis in rat brains as indicated by TUNEL staining （P0.01）. Compared with the MG， the LD， MD， and HD groups showed reduced neuronal necrosis， nuclear condensation， and vacuolar degeneration in rat brain neurons， increased Nissl body density， and reduced apoptosis （P0.01）， with significant differences among the drug groups （P0.01）. Western blot results showed that compared with the sham group， the MG group had reduced Bcl-2 and p-Akt1 （P0.01） and increased Bax and p-STAT3 （P0.01）. Compared with the MG group， the drug groups showed increased Bcl-2 and p-Akt1 （P0.01） and decreased Bax and p-STAT3 （P0.01）. There were no significant changes in total Akt1 and STAT3 protein levels among the groups. ConclusionBased on network pharmacology and experimental verification， HGWT may exert its neuroprotective effects by regulating the phosphorylation levels of Akt1 and STAT3， thereby alleviating cell apoptosis， inflammatory responses， and oxidative stress in rat brain tissue following CIRI. This provides theoretical support for the clinical treatment of CIRI.

Purpose To analyze the morphological charac-teristics of papillary thyroid carcinoma(PTC)patients with NTRK gene fusion in order to provide more important morpholog-ic evidences for molecular detection.Methods A retrospective collection of 790 cases PTC was conducted.Then the patients with NTRK gene fusion were selected.The histopathological fea-tures of PTC patients with NTRK gene fusion were compared with those of classical PTC.Results Nine cases(1.1%)of NTRK fusion positive PTC were detected,including 2 cases of NTRK1 and 7 cases of NTRK3 gene fusion.The main his-topathological features were follicular subtypes,with tumors ex-hibiting multinodular infiltration or"jumping"infiltration.The cytoplasm was associated with hyaline change.The cell morphol-ogy was slight irregularity.Conclusion The incidence of NTRK fusion is low in PTC and it tends to occur in the young group.Follicular subtype is the main characteristic histopatholo-gy,with mild tumor cells.But the ability of the invasion and metastasis is strong.Therefore,NGS detection should be per-formed for early intervention and prolonging the survival of PTC patients.

Abstract: Objective　China was certified by World Health Organization as a malaria-free country in 2021. Malaria has become a rare infectious disease, and preventing the re-transmission of imported malaria and reducing deaths are the main challenges facing China after elimination of malaria. To analyze and clarify the characteristics of imported malaria deaths, and to provide prevention and treatment recommendations for overseas workers and health care workers. Methods　The data of 17 imported malaria deaths in the analysis of malaria deaths from 2016 to 2020 by the National Severe Malaria Treatment Expert Group were collected, and the relevant clinical epidemiological data and disease course records were analyzed. Results　The 17 malaria deaths were all imported from Africa with Plasmodium falciparum infection (malarial cerebral type), with no obvious regularity in the month of onset. Among them, 16 were male patients, 5 cases with underlying diseases such as diabetes mellitus, and 10 patients were first diagnosed in a second-level or lower hospital. Excluding patients who died of respiratory cardiac arrest in ambulances, the mean time difference between first onset and malaria diagnosis in 16 patients was 6.8 days (median 5.5 days), and the mean time between first onset and antimalarial treatment was 7.4 days (median 6 days), the mean time difference from initial onset to death was 10.3 days (median 8.5 days). Excluding cases with onset abroad and unknown time of return, all 14 patients developed the disease within 30 days after returning to China. Conclusion　All the fatal cases were infected with Plasmodium falciparum imported from Africa. The patients' awareness of actively seeking medical treatment is weak, and the delay in seeking medical treatment caused by the insufficient diagnosis and treatment capacity of health institutions at the township level and below is the main reason for the deaths. It is recommended to strengthen the self-protection awareness of staff in malaria-endemic areas overseas and raise their awareness of malaria. For returnees from areas with high malaria risk, primary medical institutions should pay attention to the patient's travel history in Africa, improve the awareness of malaria diagnosis, malaria diagnosis and treatment capabilities.

Objective:To investigate the effect of growth hormone (GH) supplementation during luteal phase one cycle before ovulation induction in patients undergoing in vitro fertilization-embryo transfer (IVF-ET).Methods:IVF-ET pregnancy-assisted patients who underwent long-term Gonadotropin Releasing Hormone-agonist (GnRH-a) protocol from January 1, 2019 to June 30, 2020 were collected from the Reproductive Center of Hunan Provincial Maternal and Child Health Hospital. Among them, 106 patients (GH group) were added with GH during luteal phase one cycle before ovulation induction, and 212 patients (control group) were not added with GH. Ovulation induction and pregnancy outcome were compared between the two groups.Results:(1) There was no statistically significant difference in primary infertility/secondary infertility rate, infertility years, age, and transplant cancellation cycle rate between the two groups (all P>0.05). (2) There were no significant differences in the number of oocytes obtained, MII oocytes, two pronucleus (2PN) oocytes, high-quality embryos and average number of transplanted embryos between GH group and control group (all P>0.05). The total amount of Gn in control group and GH group was (2 109.75±555.75)IU and (1 863±610.52)IU, respectively, with statistically significant difference ( P<0.05). (3) The embryo implantation rate of the control group and GH group was 43.73%(129/295) and 60.42%(87/144), respectively, with statistically significant difference ( P<0.05). The clinical pregnancy rates of the control group and GH group were 58.79%(107/182) and 71.91%(64/89), the difference was statistically significant ( P<0.05). The spontaneous abortion rate of early pregnancy in control group (4.67%, 5/107) was slightly higher than that in GH group (3.12%, 2/64), but there was no significant statistical difference ( P>0.05). Conclusions:For patients with normal ovarian response, adding small dose of growth hormone during luteal stage one cycle before controlled hyperovulation can improve the embryo implantation rate and clinical pregnancy rate, and reduce the amount of Gn, which is beneficial to patients.

Humans ; Thyroid Cancer, Papillary ; Carcinoma, Papillary/surgery* ; Thyroid Neoplasms/surgery*

Objective:To explore effects of controlled ovarian stimulation (COS) protocols on pregnancy outcomes for patients with polycystic ovarian syndrome (PCOS) undergoing in vitro fertilization-embryo transfer (IVF-ET). Methods:A total of 1 032 patients with PCOS who underwent IVF-ET from September 1, 2016 to July 31, 2020 in the Reproductive Center of Hunan Provincial Maternal and Child Health Care Hospital were retrospectively analyzed. The patients were divided into modified long regimen group (group A, 126 cases), luteal phase long regimen group (group B, 185 cases), antagonist regimen group (group C, 344 cases), and progestin primed ovarian stimulation (PPOS) group(group D, 377 cases) according to different ovulation stimulation regimens. The ovulation promotion status [days of gonadotropin (Gn), total amount of Gn, estradiol (E 2) level on the day of human chorionic gonadotropin (HCG) injection, number of retrieved eggs, number of mature eggs (MII eggs), number of normal fertilized embryos (2PN), number of high-quality embryos] and the first frozen embryo transfer pregnancy status (clinical pregnancy rate, implantation rate, early abortion rate) were compared among the patients in each group. Results:(1) There was no significant difference in general clinical data between the four groups (all P>0.05). (2) The number of Gn days in group D was significantly less than that in groups A, B and C, and the total number of Gn was significantly less than that in groups A, B and C (all P<0.05); The E 2 level of patients in group C and group D on the day of hCG injection was significantly lower than that of group A and group B (all P<0.05); The number of eggs obtained and MII eggs in group C and group D were significantly lower than those in group A and group B (all P<0.05); The number of high-quality embryos and 2PN in group D were significantly different from those in group A, group B and group C (all P<0.05). (3) The clinical pregnancy rates of the first frozen embryo transfer after whole embryo cryopreservation in group A, group B, group C and group D were 54.72%(29/53), 56.79%(46/81), 52.56%(82/156) and 54.32%(195/359), respectively, with no significant difference among the four groups (all P>0.05). There was no significant difference in embryo implantation rate and early abortion rate among the four groups (all P>0.05). Conclusions:The modified long regimen, luteal phase long regimen, antagonist regimen and PPOS regimen can achieve better pregnancy outcomes in patients with PCOS. Among them, PPOS regimen can reduce the amount and time of Gn, and frozen embryo transfer does not affect the pregnancy outcome of patients. It can be used as one of the priority recommended strategies for PCOS patients who plan to undergo frozen embryo transfer.

Objective To investigate regional spontaneous brain activity in mild cognition impairment (MCI) patients with amnesic (aMCI) and non-amnesic (naMCI).Methods Twenty-five aMCI patients,21 naMCI patients and 15 normal controls (NC) underwent resting-state functional magnetic resonance imaging.The regional homogeneity (ReHo) map of the whole brain was obtained by calculating the similarity of each voxel with its nearest 26 voxel time series.The differences of ReHo map across the whole brain among three groups were compared.Results In aMCI group,ReHo values were lower in right frontal lobe and higher in left middle temporal gyrus and left cerebellum compared with NC (P＜0.05,Alphasim correction).In naMCI group,ReHo values were higher in anterior cingulate cortex and right middle frontal gyrus and lower in right parahippocampa gyrus,right middle temporal gyrus as well as right precuneus compared with NC (P＜0.05,Alphasim correction).Compared with naMCI,the ReHo values were significantly higher in left prefrontal gyrus,left middle temporal gyrus and lower in right cerebellum (P＜0.05,Alphasim correction).Conclusion There are differences in spontaneous brain activity of left prefrontal gyrus,left middle temporal gyrus and right cerebellum between aMCI and naMCI,which may be used to differentiate brain function between aMCI and naMCI patients.

Epigenetics is a researching hot topic of worldwide now.Increasing evidence shows that the pathogenesis of human diseases is not only influenced by the abnormalities of genetic factors but also by epigenetic mechanisms.Recent technological advances in epigenomic profiling has led to further understanding of the role epigenetic factors played in development,inflammation,aging,immunity,angiogenesis,tumorigenesis,and stem cell biology.The researchers in ophthalmology should pay close attention to the current research of major epigenetic mechanisms and their involvement in human diseases,especially ocular diseases.Moreover,the potential application of epigenetic drugs in the treatment of common human diseases also should be understood.Finally,the challenges and future perspectives underlying epigenetic research are discussed in this editorial paper.

OBJECTIVETo investigate the possibility of applying multiplex ligation-dependent probe amplification (MLPA) to the detection of azoospermia factor (AZF) microdeletion on the Y chromosome in infertile men with azoospermia or severe oligozoospermia.

METHODSDNA samples were obtained from 147 azoospermia or severe oligozoospermia patients and 154 normal controls. After denatured at 95 degrees C, the samples were hybridized to the specific probes designed for the AZF region. With the ligase, the hybrid products were amplified by a pair of universal primers labeled with FAM fluorescence, and then separated by capillary electrophoresis for data analysis. Meanwhile all the samples were subjected to multiplex-PCR (mPCR) analysis for sequence-tagged sites (STS) in the AZF region.

RESULTSSTS deletion was detected in 22 (15.0%) of the 147 patients but not in the normal controls. By MLPA, 40 (27.2%) of the patients were found with specific probe omission in the AZF region, as compared with 20 cases in the control group.

CONCLUSIONCompared with mPCR, MLPA has a better sensitivity in detecting AZF microdeletions, and it provides more precise genetic information on the AZF regions, which may contribute to in-depth exploration into the etiological mechanism of impaired spermatogenesis.

Adult ; Azoospermia ; genetics ; Case-Control Studies ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; DNA Probes ; Genetic Loci ; Humans ; Infertility, Male ; Male ; Nucleic Acid Amplification Techniques ; methods ; Oligospermia ; genetics ; Polymerase Chain Reaction ; methods ; Seminal Plasma Proteins ; genetics ; Sequence Tagged Sites ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; genetics ; Young Adult

OBJECTIVETo detect the expression of Toll-like receptor 9 (TLR9) in ovarian cancer, and to explore their clinical significance.

METHODSWestern blot method and immunohistochemical staining were used to examine the expression of TLR9 in the ovarian cancer, paracancerous tissues and normal ovarian tissues, obtained during operation from 30 ovarian carcinoma patients and 30 normal non-tumor patients. The relationships of TLR9 with pathological grade, clinical stage, and metastasis of ovarian cancer were statistically analyzed.

RESULTSThe percentage of positive cells expressing TLR9 protein in human ovarian cancer tissues, paracancerous tissues and normal ovarian tissues were 80.0%, 36.7% and 20.0%, respectively. The protein expression level of TLR9 was gradually descending (P < 0.01). The highly expressed TLR9 significantly correlated with the degree of tumor differentiation, an advanced FIGO stage and lymph node metastasis. The Western blot results showed that TLR9 protein expression in ovarian cancer, paracancerous tissues and normal ovarian tissues were 0.803 ± 0.072, 0.411 ± 0.087 and 0.113 ± 0.065, respectively. The expression of TLR9 in ovarian cancer was significantly higher than that in normal tissue and paracancerous tissues (P < 0.01).

CONCLUSIONSTLR9 has a higher expression in ovarian cancer tissues. TLR9 expression has a close relationship with pathological grades of ovarian cancer, suggesting that TLR9 plays an important role in the development and progression of ovarian cancer through immunologic mechanisms.

Adult ; Blotting, Western ; Carcinoma, Endometrioid ; metabolism ; pathology ; Case-Control Studies ; Cystadenocarcinoma, Mucinous ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Neoplasm Staging ; Ovarian Neoplasms ; metabolism ; pathology ; Ovary ; metabolism ; Toll-Like Receptor 9 ; metabolism