Volume 92  ·   Issue 11  ·  2012

Glioblastoma (GBM) is the most common highly malignant tumor of the nervous system (World Health Organization Grade IV), accounting for 50% ~55% of neuroectodermal tumors and 25% of adult intracranial tumors [1]. At present, the standard treatment of GBM is recommended, that is, concurrent chemoradiotherapy and 6 courses of temozolomide (TMZ) chemotherapy after surgery, with an average survival time of 12~15 months [2-3].

Objective To examine the antitumor activity of IL-13PE38 on solid malignant glioma cells in vitro and to investigate its relationship between the antitumor activity of IL-13PE38 and the expression level of IL-13Rα2 in malignant glioma.Methods Ten fresh tissues of anaplastic glioma and 11 fresh tissues of glioblastoma multiforme were obtained during craniotomy at First Affiliated Hospital,Wenzhou Medical College between June 2009 and December 2010.All fresh glioma cells were cultured in vitro with IL-13PE38.Then the cytotoxicity of IL-13PE38 was determined by colorimetric MTS proliferation assay and the SR (survival rate) calculated.The expression level of IL-13Rα2 was studied by immunohistochemical SABC method in 21 cases of malignant glioma.And the value of integrated optical density (IOD) was examined by computer assisted pathological image analysis system.The correlation between the IOD of IL-13Rα2 and the SR of malignant glioma cells was also studied.Results ( 1 ) There were strongly positive expression of 1L-13Rα2 in most cases ( 19/21,90％ ).And the expression level of IL-13Rα2 in glioblastoma muhiforme was higher than the expression level of IL-13Rα2 in anaplastic glioma (P ＜ 0.05).(2) As the fresh malignant glioma cells were cultured with IL-13PE38 of same concentration,the number of surviving cells decreased in different degrees.The survival rate of 14 cases were ＜ 70％ and 9 cases ＜ 50％.And the survival rate of anaplastic glioma cells was higher than that of glioblastoma multiforme cells (P＜0.05).(3) The IOD of IL-13Rα2 and the SR of malignant glioma cells were strongly negatively correlated (r =-0.093,P ＜0.01 ).Conclusion A low concentration of IL-13PE38 shows a high level of cytotoxicity for solid malignant glioma cells.And its cytotoxic efficiency depends on the expression level of IL-13Rα2.

Objective To explore the role of Rac1 in the SDF-1-induced migration and invasion of glioma cells with a specific Rac1 inhibitor.Methods Human glioma cell lines U251 treated with SDF-1 or/and specific Rac1 inhibitor were used.The migration and invasion capacities of cells in 2D cell migration/3D invasion assay were assessed.Western blot was employed to detect the levels of Rac1 and GAPDH in cell lysates and the Rac1 activity measured by Rac1 activation assays.Immunofluorescence was used to identify the expression and intracellular location of Rac1 in U251 cells.Results SDF-1 significantly increased the migration and invasion capacities of U251 cells (P ＜ 0.05 ).The stimulation of SDF-1 boosted the activity of Rac1 versus the unstimulated cells (P ＜ 0.05 ).And Rac1 was recruited to protruding edge in SDF-1-stimulated cells. Inhibition of Rac1 with specific Rac1 inhibitor decreased the migration and invasion capacities of SDF-1-induced U251 cells(P ＜ 0.05 ).In comparison with the SDF-1 treated group,the activity of Rac1 significantly decreased (P ＜ 0.05 )and the recruitment of Rac1 to protruding edge significantly decreased in the NSC23766 pre-treated group.Conclusions This study provides novel evidence that Rac1 modulates the SDF-1-induced migration and invasion of glioma cells.It suggests that the inhibition of Rac1 activation may be a new therapeutic target for glioma.

Objective To analyze the features of isocitrate dehydrogenase 1 (IDH 1 ) mutation in 165 oligodendroglial tumors.Methods IDH1 was detected in a series of 165 oligodendroglial paraffin specimens from 2009 to 2011.And their features were analyzed.Results Mutant IDH1 was detected in 111 (67.3％) tumors including 109 (98.2％ ) CGT→CAT mutations,1 (0.9％ ) CGT→AGT mutation and 1 (0.9％ ) CGT→TGT mutation.The frequencies of IDH mutation in AO,O and OA were 13/15,83.3％ and 72.9％ respectively.They were significantly higher than that in AOA (27.0％,P ＜0.001 ).Conclusion The different frequencies of IDH1 mutation in different subsets of oligodendroglial tumors may imply varied tumorigenic pathways between subsets.

Objective To explore the early changes of arterial elasticity in initial hypertension through two different methods.Methods A total of 76 hypertensive patients ( Grade 1 - 2,n =32) were assigned into hypertension group while the age and gender-matched healthy population was selected as normal group (n =44 ).All of them were monitored for baPWV in right carotid artery by wave intensity technique.The parameters of two methods were compared between two groups.Results Higher bi-lateral baPWV were found in the hypertension group( right:( 1472 ± 198 ) m/s vs ( 1353 ± 233 ) m/s),( left:(1465 ± 198) m/s vs (1353 ± 241 ) m/s)( P ＜ 0.05).Compared with the normal group,Patients in the hypertension group had statistically higher W1 ( (9.8 ±3.8) × 103 mm Hg·m·s-3 vs (7.4 ±2.4) × 103 )mm Hg·m·s-3,W2((2.8±1.1) × 103 mm Hg·m·s-3 vs (2.0 ± 1.0) x 103 mm Hg·m·s-3),stiffness parameter(β) ( (6.9 ± 2.6 ) vs ( 5.1 ± 1.3 ) ),pressure-strain elastic modulus (Ep) ( ( 99 ± 39 )kPa vs (57 ± 17) kPa),one-point wave velocity(PWVβ) ((6.0 ± 1.1 ) m/s vs (4.6 ±0.6) m/s),PWVWI((6.2±1.4) m/svs (4.4±1.0) m/s)(P＜0.01).R-W1((100±14) ms vs (103±13) ms),( W1 -W2 ) ( (261 ± 20) ms vs ( 274 ± 15 ) ms ) and arterial compliance ( AC ) ( (0.8 ± 0.4 ) mm2/kPa vs (1.1 ±0.3) mm2/kPa)in the hypertension group were statistically lower than those of the normal group (P ＜ 0.05).Conclusion The initial hypertensive patients may have impaired arterial elasticity during an early stage.

Objective To explore the therapeutic effects of posterior transpedicular screw internal fixation in atlantoaxial complex fracture patients.Methods Among 17 patients,there were Anderson Ⅱ type of odontoid plus atlas facture (n =10) and Hangman plus atlas fracture (n =7).They were categorized by their preoperative spinal cord scores according to the Frankel classification:C-class ( n =2 ),D-class ( n =6) and E-class ( n =5 ).Transpedicular screw internal fixation was performed to stabilize atlas and axis in 8 cases.Granular allograft bone was planted onto the surface of posterior arches in both atlas and axis.And the other 5 cases underwent no bony fusion.Results All patients were followed up for an average of 24 months.All screws were successfully placed in atlas and axis.And the cases of atlantoaxial dislocation were reduced.Bony fusion was observed at Months 3 - 6 post-operation.Atlantoaxial rotational function restored satisfactorily in all patients.But axial rotation was partially lost.Conclusion Posterior transpedicular screw internal fixation can regain atlantoaxial joint stability immediately and promote bony fusion.And it is an effective therapy for atlantoaxial complex fracture.

Objective To Compare the two techniques in the treatment of congenital kypho-scoliosis caused by fully-segmented hemivertebra.Methods 44 consecutive cases,21 males and 23 females.The average age at surgery was 11.4 years ranging from 2 to 17 years,20 patients underwent one-stage anterior and posterior hemivertebra resection and instrumentation.24 patients underwent one-stage posterior hemivertebra resection and instrumentation.Long cassette standing radiographs were taken before and after surgery and at the final follow-up.The apical translation,the Cobb's angle in the coronal and sagittal plane were measured and analyzed.The medical records were reviewed and the complications were recorded.Results The average follow-up was 39.2 months ranging from 24 to 72 months.The 2 groups did not demonstrate any significant differences in gender,age at surgery,preoperative and postoperative Cobb angle,blood loss,or fusion segments (all P ＞ 0.05 ).However, the anterior and posterior group demonstrated a less curve flexibility and longer operative time compared with the posterior groups ( all P ＜0.05 ).Complications:the anterior and posterior group included pedical cutting in 1 case,lumber curve decompensation in 1 case and crankshaft phenomenon in 2 cases.The posterior group included pedical cutting in 1 case and length of instrumentation in 1 case.Conclusions Two techniques are safe and effective procedure for the congenital kypho-scoliosis.Anterior and posterior hemivertebra resection is indicated to rigid deformity,and anterior epiphysiodesis is needed to avoid crankshaft phenomenon for patients at an earlier skeletal age.Posterior hemivertebra resection can save operation time and is less invasive compared with anterior-posterior approach.

Objective To investigate the Bloodless Medical and Surgical Procedures for retroperitoneal neoplasm resection.Methods Retrospectively analyse the Bloodless Medical and Surgical Procedures during 36 retroperitoneal neoplasm resections from Beijing Shijitan Hospital Affiliated to Capital Medical University from September 2009 to December 2010,to discuss the preoperative preparation,anesthetic induction and maintainance, intraoperative monitoring and use of vasoactive drugs related experience.Results All the patients were safe during the perioperative period,without any operative and anesthetic complication.By applying the Bloodless Medical and Surgical Procedures,the intraoperative haemodynamics maintained steady,the mean hematocrit decreased from 0.368 ± 0.095 before autologous blood collection to 0.252 ±0.032 before the ends of operation.Majority of the patients(91.7％ ) stop using vasoactive drugs and extubated within 1 h after operation,and return wards.Conclution Erythroeyte-raising medicine therapy and modified preservation autologous blood transfusion are important process of Bloodless Medical and Surgical Procedures.

Objective To explore the relationship of venous sinus stenosis and idiopathic intracranial hypertension (IIH) and to discuss the efficacy and strategy of endovascular treatment.Methods Balloon angioplasty and stent placement were performed in 24 IIH cases by lateral sinus stenosis.Mechanical thrombus maceration was performed before stent placing in 5 cases with suspected chronic dural sinus thrombosis.The clinical data of clinical presentations,intracranial pressure,pressure of lateral sinus,pressure gradient between distal and proximal lateral sinus stenosis,methods of treatment and therapeutic outcomes were retrospectively analyzed.Results Intracranial pressure dropped from (314 ±56) mm H2O of pre-operation to (197 ±31 ) mm H2O of post-operation.The post-treatment pressure gradient was obviously high((159 ±11 ) mm H2O)and a reduction was found ((95 ± 11) mm H2O).Headache in 16 cases,vision in 13 and papilledema in 10 were resolved or improved.There was no other permanent procedurerelated morbidity.The patients were followed up for 2-19 months.And no in-stent restenosis occurred.Conclusion Endovascular treatment is effective for IIH by venous sinus stenosis.

Objective To estimate the prevalence of visual impairment among adults aged ≥ 40 years,compare the prevalence between Uyghur and Han residents and analyze the causes of visual impairment.Methods Population-based cluster sampling survey was conducted among a target population of 5032 persons ≥ 40 years of age from Shuimogou District in Urumqi, Xinjiang Uyghur Autonomous Region.Distance visual acuity (VA) was measured with a Tumbling-E chart at 5 m.Presenting and best corrected VA were recorded.Clinical evaluations were performed.And primary causes of visual impairment was determined for those with VA ＜0.3.Results Among them,there were 4104 responders ( Uyghur 2006 persons,Han 2098 persons) with a response rate 81.56％.Prevalence of bilateral blindness was 1.25％/1.86％ (Uyghur/Han) ( P ＜ 0.05 ) and bilateral low vision 4.04％/3.29％ (Uyghur/Han) (P ＜ 0.05 ).Primary cause of visual impairment among participants was cataract ( cause proportion of 49.23％ among Uyghurs and 51.85％ among Hans).Visual impairment was correlated with advanced age,males and poor literacy.Prevalences of bilateral blindness and bilateral low vision in age group 40 - 50 yr increased from 0.48％ and 1.13％ to 3.47％ and 10.88％ in age group ≥ 70 among Uygur citizens respectively (x2 =22.36,P ＜ 0.05 vs x2 =70.97,P ＜ 0.05 ).Relative data among the Han citizens varied from 0％ and 1.42％ to 4.39％ and 7.89％ (x2 =27.94,P ＜ 0.05 vs x2 =39.99,P ＜ 0.05 ).Conclusion The most frequent cause of visual impairment in this area is cataract (Uyghur 49.23％ vs Han 51.85％ ) while retinal diseases and glaucoma are important causes as well.Blindness prevention programs targeting the elderly should be expanded,particularly in areas with a limited access and affordability of eye care services.

Objective To investigated the impact of viral load decline on virus-specific T-cell reactivity on patients with chronic hepatitis B.Methods 23 cases of patients with chronic hepatitis B were recruited randomized to therapy with nucleoside analogue or alpha interferon from January 2009 to April 2010.Peripheral blood mononuclear cells (PBMCs) were collected longitudinally at baseline and the time of HBV DNA undetected.T-cell reactivity to HBV core antigens were tested using Elispot assays and Luminex.Results (1) The frequency of T cell reactivity induced by HBcAg in patients with chronic hepatitis B were 91.3％ at the time of HBV DNA undetected,which significantly higher than The frequency of 69.6％ at baseline.The frequency between nucleoside analogue treatment group and alpha interferon treatment group was no significant difference.(2) The average response magnitude was expressed as spot forming unit (SFU) per million input cells.SFU of T cell responses to HBcAg was 120 SFU/106 PBMCs at baseline,much lower than SFU of 1060 SFU/106 PBMCs at the time of HBV DNA undetected.No significant difference between patients with negative T cell reactivity at baseline and patients with positive T cell reactivity at baseline was found.In patients with initial virological response (IVR) to therapy and patients with early virological response (EVR),no significant difference was found in the magnitude at baseline as well as at the time of HBV DNA undetected.(3) The average response magnitude of nucleoside analogue treatment group was 1713 SFU/106 PBMCs at the time the time of HBV DNA undetected,higher than 189 SFU/106 PBMCs at baseline.But in interferon treatment group,the average response magnitude was no significant difference,120 SFU/106 PBMCs at the baseline and 305 SFU/106 PBMCs at the time the time of HBV DNA undetected respectively.The average response magnitude in nucleoside analogue treatment group was greater than that in interferon treatment group.(4)As to compare difference of IFN-γ concentration in supernatant of T cell culture solution stimulated by HBcAg,IFN-γsecreted by T cell at the time of HBV DNA undetected was clearly higher than IFN-γ secreted at baseline,(38 ± 9) ng/L and (90 ± 9) ng/L respectively.Conclusions Antiviral therapy made profit to improve virus-specific T-cell reactivity in patients with chronic hepatitis B,suggesting the importance to investigate HBV specific T cell responses.

Objective To evaluate the effect of ultrasound-guided percutaneous radiofrequency ablation (RFA) in treating advanced hepatocellular carcinoma (HCC) and to analyze the prognostic factors.Methods A total of 90 patients with unresectablely advanced HCC underwent percuatenous RFA therapy between July 2000 and June 2011 were enrolled into the study.According to the 6th UICC/AJCCTNM system,80 and 10 patients were in stage Ⅲ and Ⅳ,respectively.78 patients were men and 12 patients were women.Ages ranged from 24 to 87 years old (mean ±SD,59 ± 12 years).The tumor size ranged from 1.5 to 8.0 cm ( mean ± SD,4.5 ± 1.4 cm).The maximum tumor of 73 patients ( 81.1％ ) was larger than 3.0 cm.31 patients (34.4％ ) had 2 -4 tumors.The Child-Pugh classification of B and C were 32 patients (35.6％) altogether.By regular follow-up,enhanced CT combined with AFP was used to evaluate the effect after RFA.Kapian-Meier model and Log-rank test were used in univariate analysis and Cox regression model was used in multivariate analysis to identify prognostic factors for survival.P ＜ 0.05 was considered statistically significant difference.Results Complete tumor necrosis rate after initial RFA was 90.9％ ( 120/132 tumors).Serious complications were developed in two patients (2.2％ ) and no treatment-related death occurred.3 - 129 months were followed up.Local recurrence rate was 15.2％ (20/132 tumors).The 1-,3-,5-year overall survival rates were 83.3％,48.3％,21.9％,respectively,and the median survival time was 35 months.The univariate analyses showed that patients with Child-Pugh classification of A,tumor less than 3.0 cm,applying CEUS,using standard treatment protocols,achieving complete tumor necrosis and without tumor recurrence survived longer ( P ＜ 0.05 ).Child-Pugh classification and the standard treatment protocols were identified as independent prognostic factors for survival by multivariate model ( P =0.001,P ＜0.001 ).Conclusion Paying attention to the following factors of CEUS,standard treatment protocols,initially complete tumor necrosis and liver protection therapy for patients with advanced HCC is helpful to improve the patients' survival.

Objective To detect deep-level microvascular structure in rat hind limb by synchrotron radiation and microangiographic technique.Methods Microangiography in vivo and ex vivo was performed by synchrotron radiation based absorption imaging and phase contrast imaging,with omnipaque and barium sulfate solution as contrast media,respectively,and synchrotron radiation-based micro-computed tomography (SRmCT) was also performed to reveal three-dimensional morphology of the blood vessel in rat hind limb.Results Using microangiographic technique in vivo and in vitro (with barium sulfate),blood vessels in the rat limb muscle could be visualized with high resolution,and the fourth branches of iliac artery in rat hind limb could be detected with the minimum visualized blood vessels about 40 μm and 9 μm in diameter,respectively.In addition,the vascular network could be defined and analyzed at the micrometer scale from the 3D renderings of limb vessel as shown by SRmCT.Conclusion Synchrotron radiation-based microangiography and SRmCT thus provided a practical and effective means to observe the microvasculature of rat hindlimb,which might be useful in assessment of angiogenesis in lower limbs.

Objective To determine the role of toll like receptor 4 (TLR4) in intimal hyperplasia induced by low shear stress (LSS).Methods TLR4-/- mice and control mice C57BL/6J were used.Polyethylene cuff was placed on murine carotid to establishing a LSS model.Cultured vascular endothelial cells under LSS condition were used as an in vitro LSS cell model.Intimal hyperplasia was evaluated pathologically.TLR4 was tested by Western blot and the expression of IL-1 β,1L-6 and TNF-α mRNA were detected using RT-PCR.Cell proliferation was determined by detecting DNA synthesis.Results LSS elicited significant carotid intimal hyperplasia in normal mice but a slight neointima formation in TLR4-/- mice (42.67 ± 16.46 vs 7.03 ±2.95,P ＜0.05).LSS upregulated the expression of TLR4 (2.30 ±0.66 vs 0.16±0.10,P＜0.05),as well as the mRNA of IL-1β(6.52 ±3.15 vs 1.65 ±0.45,P＜0.01),IL-6 ( 16.17 ±7.49 vs 6.50 ± 1.84,P ＜0.01 ) and TNF-α(9.98 ±3.77 vs 2.72 ± 1.03,P ＜0.01 ) in normal mice.However,only moderate increases in IL-6 and TNF-α mRNA were observed in TLR4-/- mice.LSS induced the proliferation in cultured endothelial cells.And it was further enhanced by TLR4 overexpression (177 ±33 vs 83 ± 15,P ＜0.05) but attenuated by TLR4 silencing (40 ±8 vs 83 ± 15,P＜0.05).Conclusion TLR4 plays an important role in LSS-induced intimal hyperplasia.It is likely that LSS induces the proliferation of endothelial cells through TLR4-mediated inflammatory reaction and ultimately promotes intimal hyperplasia.

Objective To explore the antiviral efficacy of small interfering RNAs (siRNAs)/shRNA targeting preC/C of HBV in human hepatoma cells Huh-7 and HepG2.2.15 cells.Methods Three 21 nucleotide(nt) siRNAs for treating HBV preC/C gere were designed and synthesized according to the HBV genome in GenBank accession numbers (U95551); simultaneously,one 21-nt-long non-homologous siRNA was also designed randomly for negative control.They were cloned into vector pU6 for constructing shRNA-expressing plasmids pU6-C1,pU6-C2,pU6-C3 and control pU6-C4. To assess the function of siRNAs,a reporter gene system was constructed.The HBV preC/C gene was synthesized by PCR with pTHBV1.3 as the template.The preC/C gene was then inserted into the enhanced green fluorescent protein expression vector (EGFP-N1) in order to construct the recombinant plasimid pEGFP-preC/C (E-C),which carries the EGFP reporter gene.The three shRNA-expressing plasmids-pU6-C1,pU6-C2,or pU6-C3-was each then cotransfected into Huh-7 cells along with either reporter gene expression vector E-C or the controls; or these three plasmids-pU6-C1,pU6-C2,or pU6-C3-was each cotransfected into HepG2.2.15 cells along with the controls.First,upon determination of the number of cells exhibiting EGFP expression in Huh-7cells as detected by an BH-2 fluorescence microscope and FACS-440 flow cytometry at different times after cotransfection,the investigators evaluated the inhibitory efficiency of the three shRNA-expressing plasmids by an EGFP reporter system in cultured cells.Subsequently,the expression amount of HBsAg and HBeAg in HepG2.2.15 cell supernatant at 24,48,72 and 96 h post-cotransfection was detected by enzymelinked immunosorbent assay (ELISA).Immunofluorescence was used to detect the expression of HBsAg and HBcAg at 72 h post-cotransfection in HepG2.2.15 cells.The copy level of HBV mRNA transcripts cDNA in HepG2.2.15 cells was further investigated through quantitative real-time polymerase chain reaction ( realtime PCR).Results In comparison with single plasmid transfection pEGFP-N1 or E-C,fluorescence microscope examination and flow cytometry detection at 48 hours after cotransfection indicated that the expression of the reporter gene EGFP in cotransfected group Huh-7 cell involving pU6-C1,pU6-C2 or pU6-C3 resulted in an 80％ reduction in EGFP signal relative to the controls ( P ＜ 0.01 ).It was also found through immunofluorescence that the expression of HBsAg and HBcAg in HepG2.2.15 cells was reduced markedly (P ＜ 0.01 ),that the copy level of HBV mRNA transcripts cDNA as detected at 48 hours after cotansfection by quantitative real-time PCR was reduced respectively by 73.9％ ± 1.2 ％ ( P =0.029 ),48.2％ ± 1.8％ and 35.8％ ± 1.4％ ( P =0.037,0.040) relative to the control,that it conformed with that detected by fluorescence microscope/flow cytometry,ELISA,and immunofluorescence ( P ＜ 0.01 ).Thereby further corroborating the antiviral efficacy of RNAi.The efficacy was obvious at 48 h,reaching a peak at 72 h.Conclusion For the first time it has been found that RNAi induced by siRNA/shRNA targeting HBV preC/C gene is effective and specific in inhibiting HBV replication and expression in human hepatoma cells Huh-7 and HepG2.2.15 cells.Our data suggest that RNAi may provide an effective,viable approach in gene therapy to treating major infectious diseases such as HBV/HCV/HIV infection.

With the improvement of living standard and the change of people's diet structure in China, the incidence of gout has increased significantly in the past 10 years. Although hyperuricemia as the pathogenesis of gout is well known and the detection of serum uric acid level is quite popular, there are still a large number of misdiagnosed cases in clinic. The purpose of this study is to retrospectively analyze the serum uric acid level and related influencing factors in patients with acute gout attack, and further analyze the significance of serum uric acid detection in acute gout attack, so as to correctly understand the diagnostic value of serum uric acid in acute gout attack.

Multiple organ dysfunction syndrome (MODS) has always been an important medical problem affecting human health and survival. When various public health emergencies occur, the key to the treatment success rate is the treatment of those patients with early MODS or multiple organ failure (MOF) due to the limitation of early treatment methods and strategies [1-2]. In hospitals, MODS patients are an important part of hospital mortality [3].

Glioma is the most common tumor of central nervous system, accounting for about 35% ~61% of intracranial tumors. According to the differentiation of tumor cells, it can be divided into astroglioma, oligodendroglioma, ependymoma, medulloblastoma, glioblastoma multiforme, etc. Glioma cells grow invasively, have no clear boundary with normal brain tissue, are highly proliferative and invasive, have high mortality, poor prognosis and short survival time, especially the median survival time of glioblastoma patients is only about 14 months [1]. At present, the main treatment of glioma is postoperative radiotherapy and chemotherapy. In recent years, new therapeutic methods such as immunotherapy and gene targeted therapy have been gradually applied in clinical practice.

The patient, a 56-year-old female from Shanxi, was admitted to the PLA General Hospital in September 2010 due to left hip discomfort for 6 years, diffuse spinal pain and limb pain for 3 years, which aggravated for 10 months. The patient began to suffer from left hip soreness and discomfort without inducement since August 2004, and the leg was heavy when walking, and the left leg needed to draw an arc outward to step forward, but it did not affect daily work and did not treat it. After 3 years, the back pain began to appear, which was severe, especially when stretching the waist and back, accompanied by tightness and stiffness of the whole body and inflexibility of the extension of the limbs. In November 2009, the pain diffused to the whole body, focusing on the back, causing the spine to not dare to straighten, and gradually appeared humpback deformity within half a year, and she was bedridden. Lumbar vertebra radiographs were taken locally, skeletal fluorosis was diagnosed, and calcium supplementation was given. The whole body pain improved for a time around 3 weeks, and she was able to stand and perform a small amount of activities. After 3 days, severe diffuse pain in the whole body appeared again without inducement, resulting in bed-ridden and unable to take care of herself.