Objective:To analyze the clinical diagnosis, treatment ，and surgical timing of otogenic intracranial complications. Methods:The clinical data of 11 patients with intracranial complications with ear symptoms as the first manifestation in Department of Otorhinolaryngology Head and Neck Surgery, Qilu Hospital of Shandong University（Qingdao） from December 2014 to June 2022 were collected, including 8 males and 3 females, aged from 4 to 69 years. All patients had complete otoendoscopy, audiology, imaging and etiology examination, and the diagnosis and treatment plan was jointly developed through multidisciplinary consultation according to the critical degree of clinical symptoms and imaging changes. Among the 11 patients, 5 cases were treated with intracranial lesions first in neurosurgery department and middle ear lesions later in otolaryngology, 3 cases of meningitis, were treated with middle ear surgery after intracranial infection control, 1 case was treated with middle ear lesions and intracranial infection simultaneously, and 2 cases were treated with sigmoid sinus and transverse sinus thrombosis conservatively. They were followed up for 1-6 years. Descriptive statistical methods were used for analysis. Results:All the 11 patients had ear varying symptoms, including ear pain, pus discharge and hearing loss, etc, and then fever appeared, headache, disturbance of consciousness, facial paralysis and other intracranial complication. Otoendoscopy showed perforation of the relaxation of the tympanic membrane in 5 cases, major perforation of the tension in 3 cases, neoplasia in the ear canal in 1 case, bulging of the tympanic membrane in 1 case, and turbidity of the tympanic membrane in 1 case. There were 4 cases of conductive hearing loss, 4 cases of mixed hearing loss and 3 cases of total deafness. Imaging examination showed cholesteatoma of the middle ear complicated with temporal lobe brain abscess in 4 cases, cerebellar abscess in 2 cases, cholesteatoma of the middle ear complicated with intracranial infection in 3 cases, and sigmoid sinus thrombophlebitis in 2 cases. In the etiological examination, 2 cases of Streptococcus pneumoniae were cultured in the pus of brain abscess and cerebrospinal fluid, and 1 case was cultured in streptococcus vestibularis, Bacteroides uniformis and Proteus mirabilis respectively. During the follow-up, 1 patient died of cardiovascular disease 3 years after discharge, and the remaining 10 patients survived. There was no recurrence of intracranial and middle ear lesions. Sigmoid sinus and transverse sinus thrombosis were significantly improved. Conclusion:Brain abscess, intracranial infection and thrombophlebitis are the most common otogenic intracranial complications, and cholesteatoma of middle ear is the most common primary disease. Timely diagnosis, multidisciplinary collaboration, accurate grasp of the timing in the treatment of primary focal and complications have improved the cure rate of the disease.
Female ; Humans ; Male ; Brain Abscess/therapy* ; Cholesteatoma ; Deafness/etiology* ; Hearing Loss/etiology* ; Lateral Sinus Thrombosis/therapy* ; Retrospective Studies ; Thrombophlebitis/therapy* ; Child, Preschool ; Child ; Adolescent ; Young Adult ; Adult ; Middle Aged ; Aged ; Cholesteatoma, Middle Ear/therapy* ; Central Nervous System Infections/therapy* ; Sinus Thrombosis, Intracranial/therapy* ; Ear Diseases/therapy*

Objective:To investigate the relationship of microcystic elongated fragmented (MELF) and clinicopathological features of patients with low grade endometrial endometrioid carcinoma, and to analyze its impact on prognosis.Methods:The clinical pathological data of 512 cases with low grade endometrial endometrioid adenocarcinoma were collected. The MELF invasive pattern in all of the sections were reappraised. The correlations between MELF pattern and clinicopathological features were analyzed by chi-square test, and the independent risk factor of lymph node metastasis were evaluated by Logistic multivariate regression analysis. Survival curves was drawn by Kaplan-Meier method, and Log-rank test was used to compare progression free survival (PFS) between patients with or without MELF pattern. Disease progression-related multivariate analysis was carried out by Cox proportional hazards model.Results:MELF pattern was observed in 12.9% (66/512) low grade endometrioid adenocarcinoma. It was significantly associated with cervical stroma invasion, more than half of the depth of myometrial invasion, lymph node metastasis and vessel invasion ( P<0.05). In addition, MELF pattern was an independent risk factor for lymph node metastasis ( P<0.05). The 5-year PFS of patients with and without MELF pattern were 95.0% and 96.0% respectively ( P>0.05). Conclusions:The patients with MELF pattern are more likely accompany with cervical stroma and deeper myometrium invasion, vessel invasion, and lymph node metastasis, and it is an independent risk factor of lymph node metastasis. However, MELF pattern has no significant impact on prognosis of patients with endometrioid carcinoma.

Objective:To investigate the relationship of microcystic elongated fragmented (MELF) and clinicopathological features of patients with low grade endometrial endometrioid carcinoma, and to analyze its impact on prognosis.Methods:The clinical pathological data of 512 cases with low grade endometrial endometrioid adenocarcinoma were collected. The MELF invasive pattern in all of the sections were reappraised. The correlations between MELF pattern and clinicopathological features were analyzed by chi-square test, and the independent risk factor of lymph node metastasis were evaluated by Logistic multivariate regression analysis. Survival curves was drawn by Kaplan-Meier method, and Log-rank test was used to compare progression free survival (PFS) between patients with or without MELF pattern. Disease progression-related multivariate analysis was carried out by Cox proportional hazards model.Results:MELF pattern was observed in 12.9% (66/512) low grade endometrioid adenocarcinoma. It was significantly associated with cervical stroma invasion, more than half of the depth of myometrial invasion, lymph node metastasis and vessel invasion ( P<0.05). In addition, MELF pattern was an independent risk factor for lymph node metastasis ( P<0.05). The 5-year PFS of patients with and without MELF pattern were 95.0% and 96.0% respectively ( P>0.05). Conclusions:The patients with MELF pattern are more likely accompany with cervical stroma and deeper myometrium invasion, vessel invasion, and lymph node metastasis, and it is an independent risk factor of lymph node metastasis. However, MELF pattern has no significant impact on prognosis of patients with endometrioid carcinoma.

Objective:To investigate the clinical management value of chitinase 3-like 1 protein(CHI3L1) in hepatocellular carcinoma (HCC) by studying the expression of CHI3L1 in peripheral blood, liver cancer and paired adjacent non-tumor tissues.Methods:Retrospective study. From 2013 to 2017, 405 patients with HCC in Third Affiliated Hospital of Naval Medical University were enrolled into the study. Meanwhile, 112 patients with liver cirrhosis (LC), 114 health subjects were included as disease and health controls. CHI3L1 in peripheral blood was detected by ELISA kit. Tissues array was made by collecting 90 pairs of tumor tissues and matched paracancer tissues, from HCC patients who were conformed by pathology. The expression of CHI3L1 in HCC tissues was analyzed by immunohistochemistry. Differences between independent groups were tested by Mann-Whitney U test or Kruskal Wallis H test, Pearson correlation analysis was used for analyzing the relationship between two subjects, and matched rank sum test was used for cancer tissue and adjacent tissue comparison. Results:The median (quartile) of CHI3L1 protein in LC group, HCC group and NC group was 195.8 (103.3,330.4) μg/L,118.2 (74.9,201.0) μg/L,46.8 (30.7,66.4) μg/L independently. The protein level of CHI3L1 in LC group was significantly higher than that in HCC group and health control group ( Z=5.186,12.928, P<0.001). HCC group was significantly higher than that in health control group ( Z=10.788, P<0.001). The level of CHI3L1 in HCC group was not related to whether liver cirrhosis was accompanied ( Z=-0.286, P=0.775). The level of serum CHI3L1 was positively correlated with noninvasive fibrosis markers (HA, PⅢNP, Ⅳ-C, FIB-4 index) ( r=0.202,0.159,0.299 and 0.221, P<0.05) and negatively correlated with ALB( r=-0.326, P<0.05) while positively correlated with AST and PT( r=0.138, 0.160, P<0.05). Positively correlation was observed between CHI3L1 and tumor size ( r=0.284, P<0.001). CNLC stage [CHI3L1 level in advanced group125.2(81.9,228.5)μg/L was higher than that in early group112.0(70.2,169.2)μg/L ( Z=-2.326, P=0.018)], but no correlation with microvascular invasion( Z=-1.531) and tumor capsule(χ 2=0.818, P>0.05). In 73 cases of HCC tissues, the positive rate of CHI3L1 was 78% (57/73) in cancer tissues and 83%(61/73) in paired adjacent non-tumor tissues. The staining intensity score of paracancer tissue 1.5(1.5,2.5) was higher than that of cancer tissue 1.5(1.5,2.0)( Z=-2.053, P=0.040). Conclusions:The tissue source of CHI3L1 protein in HCC includes cancer tissue and paracancerous tissue. The detection of serum CHI3L1 level is helpful to evaluate tumor load assessment and disease stratification management in HCC.

Objective To investigate the effect of angiotensin Ⅱ on protein kinase Cε (PKCε) and protein kinase Cα (PKCα) expression in hepatic stellate cells.Methods Hepatic stellate cell (HSC)-T6 cells were treated with different concentrations of angiotensin Ⅱ and the proliferation of HSC-T6 cells was detected by methyl thiazolyl tetrazolium (MTT) assay.The expression of PKCε and PKCα was detected by immunofluorescence staining.PKCε and PKCα mRNA levels was detected by real time polymerase chain reaction (PCR).Results Angiotensin Ⅱ concentrated the proliferation of HSC-T6 cells and the level of hydroxyproline (F =25.321,13.283,P ＜ 0.001) and showed a dose-dependent effect.With the increase of angiotensin Ⅱ concentration,PKCε significantly increased and translocated in the cell membrane;PKCα increased significantly,especially in transplanted membrane and cytoplasm (F =21.387,19.431,P ＜0.01),and showed obvious dose effect.Meanwhile,Angiotensin Ⅱ increased the expression of PKCε and PKCα,and induced cell proliferation by up-regulating PKCε and PKCα mRNA levels (F =13.279,15.174,P ＜ 0.05).Conclusions Angiotensin Ⅱ can up-regulate the expression of collagen in hepatic stellate cells in a dose-dependent manner,increase the expression of protein kinase Cε and Cα,and promote the proliferation of hepatic stellate cells.

Objective To investigate the regulation of interferon α-1b (IFNα-1b) on protein kinase Cε(PKCε) and protein kinase Cα(PKCα) which inhibit the fibrosis of hepatic stellate cells (HSC) ,and to explore its mechanism .Methods HSC-T6 cells were treated with different levels of IFNα-1b (100 , 200 ,400 ,800 and 1000 U/mL) and the proliferation of HSC-T6 cells was analyzed by methyl thiazol tetrazolium (MTT) assay .Changes of hydroxyproline level were analyzed .The expressions of PKCεand PKCαwere detected by immunofluorescence staining . PKCε, PKCα,β-catenin and Survivin mRNA levels were detected by RT-PCR . PKCε, PKCα,β-catenin and Survivin protein levels were detected by Western blot . Variance analysis was conducted by using one-way ANOVA approach . Results The inhibition rates of 100 , 200 , 400 , 800 and 1000 U/mL IFNα-1b treatment after 24 hours of administration were (15 .85 ± 1 .05)％ ,(36 .59 ± 1 .03)％ ,(45 .12 ± 1 .05)％ ,(50 .00 ± 1 .01)％ and (62 .20 ± 1 .02)％ ,respectively ,with statistically significant differences among groups (F=27 .478 , P<0 .01) .The 48h inhibition rates were (20 .87 ± 1 .09)％ ,(43 .96 ± 1 .08)％ ,(53 .85 ± 1 .08)％ ,(64 .84 ± 1 .06)％ and (74 .72 ± 1 .07)％ ,respectively ,with statistically significant differences among groups (F=25 .321 , P< 0 .01 ) . half maximal inhibitory concentration at 48 h was 343 .47 U/mL . The levels of hydroxyproline in 100 ,200 and 400 U/mL IFNα-1b groups were (7 .48 ± 0 .28) ,(6 .26 ± 0 .17) and (3 .86 ± 0 .20) μg/mL ,respectively ,which were lower than that in control group (8 .47 ± 0 .32) μg/mL .The differences were all statistically significant (t=4 .033 ,10 .564 and 21 .160 ,respective ,all P<0 .05) .The fluorescence intensities of PKCεin 100 ,200 and 400 U/mL IFNα-1b groups were all lower than that of control group .The differences were statistically significant (t=1 .984 ,2 .457 and 7 .771 ,respectively ,all P<0 .05) .The fluorescence intensities of PKCαwere also significantly lower than that of control group (t=9 .232 ,15 .921 and 22 .222 ,respectively ,all P< 0 .01) .With the increase of IFNα-1b level ,the levels of HSC-T6 PKCε,PKCα,β-catenin and survivin were significantly lower than those of control group (t=7 .020 ,24 .562 ,45 .701 and 14 .241 ,respectively ,all P<0 .01) .With the increase of IFNα-1b ,the levels of HSC-T6 PKCε,PKCα,β-catenin and survivin were significantly lower than those of control group (t=9 .564 ,4 .409 ,10 .036 and 6 .794 ,respectively ,all P<0 .01) .Conclusions IFNα-1b can down-regulate the expression of collagen in hepatic stellate cells in a dose-dependent manner ,reduce the expressions of PKCε,PKCα,β-catenin and Survivin ,and inhibit the proliferation of HSC-T6 hepatic stellate cells .

3-Hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR), the first enzyme of mavalonic acid pathway, is one of the key devices involved in ginsenoside biosynthesis based on synthetic biology approach. The open reading frame of a novel HMGR gene from Panax ginseng (PgHMGR2) was cloned and analyzed in this study. PgHMGR2-encoding protein showed 71.6% sequence similarity to a P. ginseng HMGR in GenBank. The full-length cDNA sequence of PgHMGR2 containing 1 770 bp, which encodes 589 amino acids, was cloned by RT-PCR strategy from P. ginseng. The bioinformatic analysis showed that PgHMGR2-encoding protein contained two transmembrane regions and the HMG_CoA_reductase domain, without signal peptide. The protein sequence of PgHMGR2 had the highest sequence similarity (99%) with Panax quinquefolius HMGR (GenBank accession No. ACV65036). The expression level of PgHMGR2 was the highest in flower based on a real-time PCR analysis, followed by leaf and root, and the lowest was in stem. The result will provide a foundation for exploring the molecular function of PgHMGR2 involved in ginsenoside biosynthesis based on synthetic biology approach in P. ginseng plants.

Research on medicinal model organism is one of the core technologies to promote the modernization of traditional Chinese medicine (TCM). The research progress of Salvia miltiorrhiza as medicinal model plant is summarized in this paper. The genome of S. miltiorrhiza is small and its life cycle is short, as well as this plant can be stably genetically transformed. Because S. miltiorrhiza possesses the important medicinal and economic values, recently the transcriptome and genome of S. miltiorrhiza have been significantly recovered. The research prospect of S. miltiorrhiza as medicinal model plant in TCM was discussed, including biosynthesis of active components and their genetic regulation, relationship between quality of TCM and ecological environments, and selective breeding of good quality lines. Furthermore, as medicinal model plant, the construction of mutant library for S. miltiorrhiza, the genome map with high quality, and the functional genome should be investigated. Accompanying modern investigation of life sciences, the platform for medicinal model plant, S. miltiorrhiza, will be promoted to be established. It is important to develop the ethnopharmacology and new drugs around the world.

Objective To investigate the therapeutic effect of frozen-thawed murine islets which were transplanted into diabetic rats after cultured with hyperbaric oxygenated rotary cell culture system (HORCCS). Methods The purified rat islets were divided into two groups: A. In vitro experiment groups (IvEG) : The rat islets in each subgroup were cultured in HORCCS or common medium for 30 days, then evaluated for the intracellular DNA and insulin contents of islets, and the viability and insulin secreting level of islets. B. Islet transplantation experimental groups (TxEG) : The frozen-thawed islets were cultured in HORCCS or common medium for 7 days, and then transplanted into the recipients. We observed the blood glucose level (BGL) and insulin secreting level in the recipients as well as the uhrastructure change of islets in TxEG. Results The viability and insulin secreting level of islets cultured with HORCCS at 14th day were much higher than those cultured with common medium (P <0.05). The blood glucose level in recipients transplanted with islets cultured with HORCCS recovered to normal value at the 2nd week and lasted for 8 weeks. All these recipients maintained the normal glucose tolerance curve. Electronic microscopy found microchannel outlets on the surface of the frozen-thawed islets cultured with HORCCS. Conclusions Frozen-thawed islets cultured with HORCCS could establish nutrient transmission microchannels, which were not only capable of oxygen and nutrients transmission, but also improving cryopreservation solution to diffuse inside the islet cells evenly and uniformly. So this method not only lessens islet damage from cryopreservation, but also improves the effect of transplantation.

OBJECTIVETo study the effect of xenotransplantation with pig parathyroid cells, which was prepared using cell microencapsulation technique, on the treatment of hypoparathyroidism in rats without immunosuppressor.

METHODSParathyroid cells were isolated from 10 healthy newborn pigs and encapsulated in alginate-polylysine-alginate (APA) membranes. Thirty-two aparathyroid Wistar rats were randomly allocated to microcapsule, non-microcapsule, empty microcapsule, and control groups. Each rat was injected intraperitoneally with encapsulated porcine parathyroid cells, free porcine parathyroid cells, empty capsules or 0.9% NaCl, respectively. Total serum calcium and parathyroid hormone levels were monitored continuously for 40 weeks. And then, the transplant beds were retrieved and subjected to morphologic and electron microscopic examination.

RESULTSIn those animals xenotransplanted with microencapsulated porcine parathyroid cells, the calcium and PTH levels were consistently within the normal range during the 40 weeks. In contrast, no therapeutic effects were observed in rats in the non-microcapsule group. Furthermore, neither empty capsules nor 0.9% NaCl were shown to have any effect on the recipient's serum calcium or PTH levels. After 40 weeks, electron microscopic examination demonstrated that the parathyroid cells within the microcapsules had survived well in vivo.

CONCLUSIONSXenotransplantation of microencapsulated newborn pig parathyroid cells can successfully treat hypoparathyroidism in rats without using immunosuppressive drugs. The results of this study show the possible clinical use of microencapsulated porcine parathyroid cells.

Animals ; Animals, Newborn ; Calcium ; blood ; Capsules ; Hypothyroidism ; therapy ; Parathyroid Glands ; cytology ; transplantation ; Parathyroid Hormone ; blood ; Random Allocation ; Rats ; Rats, Wistar ; Swine ; Transplantation, Heterologous ; methods