Transmembrane protein 33(TMEM33)is up-regulated in the majority of cancers.To change the expression of TMEM33 can significantly affect the proliferation,migration,and invasion of renal and cervical cancer cells.Its mechanisms of action may involve four aspects:1)participation in lipid metabolism;2)regu-lation of endoplasmic reticulum;3)regulation of intracellular calcium homeostasis;4)participation in angio-genesis.

Objective This study aimed to explore the expression pattern of transmembrane protein 33(TMEM33)in lung adenocarcinoma and its correlation with clinical pathological features.Methods Bioinformatics tools and public databases(e.g.,TCGA and GEO)were used to analyze TMEM33 expression data in lung cancer.Then,immunoblotting and real-time quantitative PCR were performed to detect TMEM33 protein and mRNA levels in four cell lines.Immunofluorescence and immunohistochemistry were also used to assess TMEM33 expression and localization in lung adenocarcinoma and normal lung tissue samples.Results Bioinformatics analysis revealed higher TMEM33 expression in lung adenocarcinoma than in normal lung tissue(P＜0.05)and a significant correlation between TMEM33 and SLC30A9 expression(P＜0.0001).Logistic regression analysis indicated an association between TMEM33 expression and tumor T stage(OR=0.48,P=0.044).Experimental results showed higher TMEM33 protein(P＜0.01)and mRNA(P＜0.001)levels in lung adenocarcinoma cell lines than in normal lung epithelial cells.Similarly,TMEM33 protein(P＜0.05)and mRNA(P＜0.01)levels were higher in lung adenocarcinoma tissues than in adjacent tissues.Immunohistochemistry confirmed high TMEM33 expression in lung adenocarcinoma tissues.Conclusion TMEM33 is highly expressed in lung adenocarcinoma,associated with malignancy and T stage,and may be a potential prognostic and therapeutic target.

OBJECTIVE: To explore the pathogenesis of two siblings (including a fetus) from a pedigree affected with Joubert syndrome. METHODS: Peripheral blood samples of the proband and his parents as well as amniotic fluid and abortion tissues of the fetus were collected. Part of the samples were used for the extraction of DNA, and whole exome sequencing (WES) was carried out to screen potential variants in the proband and his parents. Suspected variants were subjected to bioinformatics analysis with consideration of the clinical phenotype, and were verified by Sanger sequencing of the proband, fetus and their parents.The remainders were used for the extraction of RNA, and the mechanism of splicing variant was validated by reverse transcription-PCR (RT-PCR). RESULTS: WES showed that both patients have carried c.175C>T (p.R59X) and c.553+1G>A compound heterozygous variants of the TMEM237 gene. Among these, c.175C>T was a nonsense mutation inherited from the asymptomatic mother, while c.553+1G>A was an alternative splicing mutation inherited from the asymptomatic father. RT-PCR showed that this variant has resulted in aberrant splicing by exon skipping. CONCLUSION The compound heterozygous variants of the TMEM237 gene probably underlay the etiology of Joubert syndrome in this pedigree. Above finding has enriched the phenotype and variant spectrum of the TMEM237 gene, and facilitated genetic counseling and prenatal diagnosis for the family.
Abnormalities, Multiple/genetics* ; Cerebellum/abnormalities* ; Eye Abnormalities ; Female ; Genotype ; Humans ; Kidney Diseases, Cystic ; Mutation ; Pedigree ; Phenotype ; Pregnancy ; Retina/abnormalities*

BACKGROUND:Transplanted neural stem cel s can survive, proliferate and differentiate into neurons and/or glial cel s in the host, thereby promoting partial function recovery in the host.
OBJECTIVE:To study the therapeutic effects of neural stem cel transplantation on cerebral palsy rats. METHODS:Twenty-four rats were randomly divided into three groups:control group, model group and transplantation group. Animal models of cerebral palsy were made in the latter two groups. One week after modeling, rats in the transplantation group were injected 1 mL stem cel suspension (1×105) via the jugular vein, and rats in the control and model group were given the same volume of normal saline. Toe distance, step length and elevated body swing test in rats were detected, and histopathological changes in the rat brain were observed 3 weeks after transplantation.
RESULTS AND CONCLUSION:In the model group, the toe distance and step length of the front left palm were significantly lower than those of the front right palm (P<0.05). Compared with the model group, the toe distance and step length in the transplantation and control groups were significantly increased (P<0.05). In the elevated body swing test, rats in the model group presented with asymmetric swing of the
body, but rats in the other two groups exhibited symmetric swing of the body (P<0.05). Additional y, the ratio of right to left hemispheric areas was significantly higher in the transplantation and control groups compared with the model group (P<0.05). In conclusion, neural stem cel transplantation via the jugular vein can improve brain function and restore motor function in rats with cerebral palsy.

OBJECTIVE:To observe clinical efficacy and safety of naloxone hydrochloride combined with Danshen injection in the treatment of neonatal hypoxic ischemic encephalopathy(HIE). METHODS:104 patients with HIE were randomly divided into observation group and control group with 52 cases in each group. The control group was treated with Naloxone hydrochloride injec-tion 0.02 mg/kg,ivgtt,qd and conventional symptomatic treatment;while observation group was additionally treated with Danshen injection 4-6 ml+10% Glucose injection 20 ml,ivgtt,qd,on the basis of the control group. Treatment course of 2 groups lasted for 7 d. Clinical efficacy,symptom and sign recovery of 2 groups were compared,as well as levels of serum MMP-9,IL-6 and TNF-α,cerebral hemodynamic parameters and ADR. RESULTS:There was no statistical significance in MMP-9,IL-6,TNF-α and cerebral hemodynamic parameters before treatment(P>0.05). The total effective rate of observation group(92.31%)was signifi-cantly higher than that of control group (75.00%);after treatment,the recovery time of consciousness disturbance,primitive re-flex and muscle tension in the observation group were shorter than those in the control group;the levels of serum MMP-9,IL-6 and TNF-α in observation group were significantly lower than those in control group;peak systolic flow velocity(PSFV),end-dia-stolic flow velocity (EDTV) were higher in the observation group than in the control group,with statistical significance (P<0.05);resistance index (RI) was lower than in control group,but without statistical significance (P>0.05). No ADR was ob-served in 2 groups. CONCLUSIONS:Naloxone hydrochloride combined with Danshen injection can significantly promote HIE and signs recovery,reduce the levels of serum MMP-9,IL-6 and TNF-α,and improve hemodynamic parameters with good safety.

Objective To investigate the effect of sulforaphane synergistic with lithium chloride on neuroprotective of rats with traumatic brain injury. Methods According to feeney method free fall production models of traumatic brain injury induced injury models, and rats were divided into control group, model group, SFN group and collaborative group, each group 15 rats, control group without impact experiment, after operation the SFN group received daily intraperitoneal injection of sulforaphane, collaborative group were given sulforaphane and chloride lithium intraperitoneal injection, the model group received daily intraperitoneal injection of saline, the degree of nerve function impairment score (mNSS) in rats of each group postoperative 1 d, 3 d, 5 d and 7 d, and brain tissue of rats, the water content of brain tissue, HE staining of brain tissue and activity of SOD, IL-6, TNF-alpha, the level of MDA in each group postoperative 7 d were compared. Results Compared with the control group, mNSS scores in the model group, SFN group, collaborative group at each time point were significantly increased (P<0.05); After 5 d, 7 d of operation, synergy scores in SFN group and mNSS group rats were significantly lower than those in the model group (P<0.05), and mNSS score in collaborative group was significantly lower than that of SFN group(P<0.05); After 7 d of operation, the brain tissue water content in control group of rats was (69.29±2.06)%, the model group was(75.40±1.73)%, SFN group was (73.08±1.06)%, collaborative group was (71.27±1.52)%; Brain tissue HE staining results showed that the nerve cell injury of rats in SFN group and cooperative group was obviously alleviated than those in the model group, necrotic cells decreased, and the collaborative group relieve neuronal injury was the most remarkable; The SOD in model group, SFN group, collaborative group decreased significantly compared with the control group (P<0.05), IL-6, TNF-alpha, MDA levels increased significantly(P<0.05);The activity of SOD in SFN group and collaborative group increased significantly compared with the model group(P<0.05), IL-6, TNF-alpha, MDA levels decreased significantly (P<0.05), and collaborative TNF-alpha and IL-6 levels were significantly lower than that of SFN group (P<0.05). Conclusion Sulforaphane in combination with lithium chloride have remarkable neuroprotection on traumatic brain injury in rats, its mechanism may be related with the reduction of brain edema in rats, nerve cell damage, inhibition of oxidative stress and related to the release of inflammatory factors.

Objective To investigate the clinical efficacy and treatment method for no midline shift-severe craniocerebral trauma accompanied with post-traumatic acute diffuse brain swelling (PADBS).Methods 60 PADBS patients were randomly divided into conservative treatment group and operation group,30 patients in each group.The operation group was treated with intracranial pressure monitoring by implantation of the probe and decompressive craniectomy,while the conservative treatment group received conservative treatment.The postoperative recovery was observed.Results The GCS scores of operation group postoperative 7d and 15d were (11.21 ± 2.24) and (12.88 ±2.31),which were obviously higher than (7.47 ± 1.51) and (8.19 ± 1.28) of the conservative treatment group (t =2.215,2.321,all P ＜ 0.05).Postoperative long-term follow-up results indicated that,according to GOS score,63.3％ patients in the operation group recovered well,which was significantly higher than 26.7％ in the conservative treatment group.While the percent of patients with coma or dead was 6.7％ and 10.0％ in the operation group,which were significantly lower than the conservative treatment group (x2 =15.721,4.172,3.84,all P ＜ 0.05).Conclusion In general,PADBS could not be cured easliy,the operation methods of using intracranial pressure monitoring and decompressive craniectomy based on conservative treatment could help to evaluate the trauma objectivly,detect the changes of disease earlier,treat in time and assess the prognosis accurately,all which would reduce the mortality.

Five groups were assigned to include intrahepatic cholangiocarcinoma ( ICC, n=30 ) , liver cirrhosis (LC,n=30),metastatic carcinoma (MCA,n=30) and 30 healthy subjects.The serum level of GPC3 was measured by a sandwich method of enzyme-linked immunosorbent assay ( ELISA ) and alpha-fetoprotein (AFP) by microparticle enzyme immunoassay (MEIA).The serum levels of GPC3 and AFP were significantly higher than those of other groups (P<0.05).At a cut-off value of 3.5μg/L,the sensitivity and specificity of GPC3 in the diagnosis of HCC was 83.3%and 76.7%respectively.The sensitivity of combined measurement of GPC3 and AFP was better than GPC3 or AFP alone.Detectable GPC3 was significantly correlated with the presence of viral hepatitis markers and tumor size.However there was no obvious difference in tumor thrombi in portal vein ( PVTT), tumor number, age, gender or hepatic function of HCC.Thus,as a sensitive serum diagnostic marker for HCC ,GPC3 may be a good supplement to AFP in differentiating HCC from non-malignant chronic liver diseases and other liver cancers.

Objective: To observe the safety and clinical efficacy of tumor antigen-pulsed dendritic cell(DC) vaccine in treatment of advanced malignant tumor.Methods: Ninety-one patients with non-small cell lung cancer,colon and rectal cancer,melanoma,renal carcinoma,breast cancer and other malignant tumors were enrolled in this study.All patients met the selecting standard and signed informed consent.Human dendritic cells were obtained from peripheral blood monocytes by culturing them with granulocyte macrophage-colony stimulating factor and interleukin-4.DC vaccine was prepared from tumor antigen pulsed immature dendritic cells in vitro.Patients received the vaccine therapy once every week and one cycle was defined as once every week for 3 weeks.Results: All the patients received 96 cycles of DC vaccine treatment.Symptoms of toxicity included fever,shivering,aching pain of muscle,asthenia,itching,stifle and transient fatigue;most of the symptoms automatically recovered.Clinical efficacy of the treatment was evaluated in 76 patients.Thirty-one of the 76 patients were stable after treatment and 45 were in progressive situation,with the clinical benefiting rate being 40.8%.Eighty-five patients were followed up.The median time for progression was 2.6 months;the overall survival time was 0.9-30.6 months;and the median survival period was 4.5 months,with the one year survival rate being 9.2%.Conclusion: The results suggest that the DC vaccine therapy is well tolerated in treating patients with advanced malignant tumors and has satisfactory clinical benefit;the clinical value of DC vaccine therapy needs to be further observed.